bluuebunny/crossref_metadata_2025 huggingface dataset:
Adenocarcinoma arising in association with gastric heterotopic pancreas: A case report and review of the literature
Authors: Emerson Lyska , Layfield Lester J. , Rohr L. Ralph , Dayton Merril T.
Published: June 2004

We report a case of adenocarcinoma arising in a focus of heterotopic pancreas, occurring in the stomach of a 52‐year‐old man. The patient presented with gastric outlet obstruction. Radiographic studies revealed thickening of the gastric wall, but endoscopy failed to reveal a mucosal abnormality. A 50% distal gastrectomy was performed, along with vagotomy. Microscopic examination revealed extensive involvement of the muscularis propria of the distal stomach by heterotopic pancreas. The ectopic pancreas had a microscopic appearance consistent with Heinrich's class III, in which the majority of the heterotopic pancreas was characterized by cystically dilated duct structures. Occasional islets were present. Intimately associated with the cystically dilated ducts was a prominent number of small infiltrating ducts lined by columnar or cuboidal cells with enlarged hyperchromatic nuclei containing prominent nucleoli. These were consistent with a well‐differentiated invasive adenocarcinoma. Despite multiple sectioning, no connection between the adenocarcinoma and the overlying gastric mucosa was seen. Adenocarcinoma arising within ectopic pancreas is a rare occurrence with fewer than 30 well‐documented cases reported in the world literature to our knowledge. J. Surg. Oncol. 2004;87:53–57. © 2004 Wiley‐Liss, Inc.
Diffusion and Reaction of Methylaluminoxane (MAO) within a Silica Membrane
Authors: Hansen Eddy W. , Bernauer Bohumil , Blom Richard
Published: June 2004

Summary: The self‐diffusion coefficient of MAO confined within a silica membrane is not experimentally accessible by traditional PFG NMR measurements, probably due to a susceptibility effect resulting in rather broad, unobservable resonance lines. A more indirect approach was therefore applied to probe the diffusivity of MAO within a silica membrane. MAO confined in a bulk toluene solution on one side of the membrane was free to diffuse through the membrane (saturated with toluene) and into a corresponding deuterated toluene solution on the other side of the membrane where its proton NMR signal intensity was monitored as a function of time. By introducing a relevant diffusion model, the diffusivities of both toluene and MAO were derived by model‐fitting. By refining the diffusion model, the formation of methane within the silica was explained, and its diffusivity estimated. The results suggest a significant reduction in diffusivities of both MAO and toluene within the silica as compared to the corresponding diffusivities in bulk solution.

1H NMR spectra as a function of time within the shift region δ = 2.0–2.5 ppm (toluene) acquired during the first 40 h of the diffusion process.

magnified image

1H NMR spectra as a function of time within the shift region δ = 2.0–2.5 ppm (toluene) acquired during the first 40 h of the diffusion process.
Segmented Multifunctional Poly(ether ester) Polymers Containing H‐Bonding Units. Preparation and Characterization
Authors: Signori Francesca , Solaro Roberto , Lips Priscilla A. M. , Dijkstra Pieter J. , Feijen Jan , Chiellini Emo
Published: June 2004

Summary: A series of poly(ether ester)s containing amide and carbamate groups as H‐bonding units and 13–50 mol‐% of poly(ethylene glycol) (PEG) segments were prepared by polycondensation in bulk using Ti(OBu)4 as a catalyst. The copolymers were obtained starting from PEG/1,4‐butanediol mixtures and a synthetic monomer carrying H‐bonding groups. These polymers were designed for biomedical applications, where material biodegradability is required. The influence of the nature of the H‐bonding units, the length of the polymethylene spacer between the H‐bonding groups and the PEG content on the thermal and solubility properties of the copolymers was investigated. Amide‐containing copolymers were more thermally stable than those containing carbamate groups. The PEG content also slightly affected the polymer thermal stability. The DSC traces of all samples presented multiple transitions, whose shape and peak temperature were strongly dependent on the PEG content. Polymer hydrophilicity, surface free energy and equilibrium swelling in phosphate buffer solution (PBS) at 37 °C were mainly influenced by the PEG content, whereas the nature of the H‐bonding groups had little effect.

magnified image
Inflammatory malignant fibrous histiocytomas and dedifferentiated liposarcomas: histological review, genomic profile, and MDM2 and CDK4 status favour a single entity
Authors: Coindre Jean‐Michel , Hostein Isabelle , Maire Georges , Derré Josette , Guillou Louis , Leroux Agnès , Ghnassia Jean‐Pierre , Collin Françoise , Pedeutour Florence , Aurias Alain
Published: June 2004

Inflammatory malignant fibrous histiocytoma (inflammatory MFH) is a very rare tumour that occurs most often in the retroperitoneum. So far, it has been considered to be a special subtype of MFH. As it is now widely accepted that most retroperitoneal pleomorphic MFHs are dedifferentiated liposarcomas, the present study compared histological features, genomic profile (CGH analysis), and MDM2 and CDK4 status (immunohistochemistry, FISH, and quantitative PCR) in inflammatory MFHs from 12 patients and dedifferentiated liposarcomas that had an inflammatory MFH component from eight patients. Metaphase cytogenetic and FISH analyses were also performed on one inflammatory MFH. Histological review showed areas of well‐differentiated liposarcoma in nine inflammatory MFHs. CGH analysis showed 12q13–15 amplification or gain in six of seven inflammatory MFHs and in seven of seven dedifferentiated liposarcomas. Immunohistochemistry showed positivity of tumour cells for MDM2 in every tumour in both groups and for CDK4 in ten and seven inflammatory MFHs and dedifferentiated liposarcomas, respectively. Metaphase cytogenetic and FISH analysis performed on one inflammatory MFH showed the presence of a supernumerary large marker chromosome and ring chromosome with high‐level amplification of both MDM2 and CDK4 genes. FISH analysis on paraffin wax‐embedded sections showed amplifications of MDM2 and CDK4 in seven of seven inflammatory MFHs and in seven of seven dedifferentiated liposarcomas. Quantitative PCR showed amplification of MDM2 in six and of CDK4 in seven of nine inflammatory MFHs. In conclusion, this study strongly suggests that most so‐called inflammatory MFHs are dedifferentiated liposarcomas. Copyright © 2004 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
Protein expression profiles in adenocarcinomas and squamous cell carcinomas of the lung generated using tissue microarrays
Authors: Ullmann Reinhard , Morbini Patrizia , Halbwedl Iris , Bongiovanni Massimo , Gogg‐Kammerer Margit , Papotti Mauro , Gabor Sabine , Renner Heiko , Popper Helmut Hans
Published: June 2004

With the appearance of defect‐targeted therapies, the definition of tumour protein expression profiles has gained increasing importance. Two lung carcinoma tissue microarrays, one including 75 primary adenocarcinomas (ACs) and the other comprising 67 primary squamous cell carcinomas (SQCCs), were generated in the present study. On both arrays, each tumour was represented by an average of five cores. In addition, one punch of normal lung parenchyma adjacent to each tumour was included in the array. Immunohistochemical expression of 86 proteins was evaluated and the results were analysed by non‐parametric tests, hierarchical clustering, and principal component analysis. In both tumour entities, parenchyma and tumours were clearly separated by hierarchical clustering. By the same statistical approach, it was possible to distinguish ACs from SQCCs with 98% accuracy and to distinguish parenchyma adjacent to ACs from that adjacent to SQCCs with 96% accuracy. It was also possible to separate ACs into three groups that significantly differed in survival. Cathepsin E and hsp105 were identified as previously unknown predictors of survival in lung AC. In summary, this study has shown that protein profiles are feasible tools for anticipating biological behaviour. Copyright © 2004 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.
Effect of high‐fat diet on the expression of proteins in muscle, adipose tissues, and liver of C57BL/6 mice
Authors: Schmid Gerhard M. , Converset Véronique , Walter Nadia , Sennitt Matthew V. , Leung Kit‐Yi , Byers Helen , Ward Malcolm , Hochstrasser Denis F. , Cawthorne Michael A. , Sanchez Jean‐Charles
Published: June 2004

In the present study, the effect of a high fat diet on the expression of proteins in insulin target tissues was analyzed using a proteomic approach. Gastrocnemius muscle, white and brown adipose tissue, and liver were taken from C57BL/6 mice either fed on a high‐fat or a chow diet. Expression levels of approximately 10 000 polypeptides for all the four tissues were assessed by two‐dimensional gel electrophoresis (2‐DE). Computer‐assisted image analysis allowed the detection of 50 significantly (p < 0.05) differentially expressed proteins between obese and lean mice. Interestingly, more than half of these proteins were detected in the brown adipose tissue. The differentially expressed proteins were identified by tandem mass spectrometry. Several stress and redox proteins were modulated in response to the high‐fat diet. A key glycolytic enzyme was found to be downregulated in adipose tissues and muscle, suggesting that at elevated plasma fatty acid concentrations, fatty acids compete with glucose as an oxidative fuel source. Furthermore, in brown adipose tissue there were significant changes in mitochondrial enzymes involved in the Krebs tricarboxylic acid (TCA) cycle and in the respiratory chain in response to the high‐fat diet. The brown adipose tissue is an energy‐dissipating tissue. Our data suggest that the high‐fat diet treated mice were increasing energy expenditure to defend against weight gain.
Identification of post‐mortem cerebrospinal fluid proteins as potential biomarkers of ischemia and neurodegeneration
Authors: Lescuyer Pierre , Allard Laure , Zimmermann‐Ivol Catherine G. , Burgess Jennifer A. , Hughes‐Frutiger Séverine , Burkhard Pierre R. , Sanchez Jean‐Charles , Hochstrasser Denis F.
Published: June 2004

Only few biological markers are currently available for the routine diagnosis of brain damage‐related disorders including cerebrovascular, dementia, and other neurodegenerative diseases. In this study, post‐mortem cerebrospinal fluid samples were used as a model of massive brain insult to identify new markers potentially relevant for neurodegeneration. The protein pattern of this sample was compared to the one of cerebrospinal fluid from healthy subjects by two‐dimensional gel electrophoresis. Using gel imaging, N‐terminal microsequencing, mass spectrometry, and immunodetection techniques, we identified 13 differentially expressed proteins. Most of these proteins have been previously reported to be somehow associated with brain destruction or with the molecular mechanisms underlying certain neurodegenerative conditions. These data indicate that the identified proteins indeed represent potential biomarkers of brain damage. We recently showed that H‐FABP, a protein highly homologous to E‐FABP and A‐FABP identified in this study, is a potential marker of Creutzfeldt‐Jakob disease and stroke.
Chain transfer in ethylene polymerization. VII. Very reactive and depletable transfer agents
Authors: Mortimer George A.
Published: June 2004

The study of chain transfer in the free‐radical polymerization of ethylene was extended to very reactive transfer agents, particularly aldehydes and mercaptans The chaintransfer constant Cs for aldehydes was insensitive to temperature changes but was strongly reduced as pressure was increased. Mercaptans were found to deplete during polymerization (Cs > 1.0). The conventional Mayo equation was integrated to allow accurate calculation of Cs for depleting transfer agents.
Anionic polymerization of methyl methacrylate with Cr(C5H7O2)3–Al(C2H5)3 catalyst system
Authors: Deshpande A. B. , Kale S. M. , Kapur S. L.
Published: June 2004

A homogeneous catalyst system, Cr(C5H7O2)3–Al(C2H5)3, was used for the polymerization of methyl methacrylate. The yield of polymer increased up to an Al/Cr ratio of 12 and thereafter remained almost constant with increasing Al/Cr. The rate of polymerization increased linearly with increasing catalyst and monomer concentrations at Al/Cr = 12. The molecular weight, however, decreased with increasing catalyst concentration and increased with increasing monomer concentration, indicating anionic polymerization reaction. NMR studies of the polymers indicated the presence of a stereoblock structure, which changed to heteroblock structure in presence of triethylamine and hydroquinone as additives in the catalyst. In the light of these observations, the mechanism of the polymerization is discussed.
Cycloolefin polymerization initiated by transition metal–π‐allylic complexes
Authors: Kormer V. A. , Poletayeva I. A. , Yufa T. L.
Published: June 2004

A new class of catalysts for polymerization of cycloalkenes was developed, based on the interaction of these hydrocarbons with π‐allylic transition metal complexes. The reaction route (decyclization or double‐bond opening) and the microstructure of polyalkenamers obtained is determined primarily by the nature of the transition metal and also by the nature and the number of ligands bound to the metal. π‐Allylic complexes of zirconium and chromium catalyze the polymerization of cycloalkenes in both directions. For complexes of the group VI metals the substitution of molybdenum or tungsten for chromium results in complete suppression of the double‐bond opening. In the presence of Group VIII metal compounds the polymerization of cycloolefins occurs exclusively at the double bond. The introduction of acidic ligands into the internal sphere of the π‐allylic compound does not affect the reaction route but results in a significant rise of the yield of polyalkenamers and enriches the content of trans‐1,4‐units. Lewis acids act as activators for complexes of all the metals, the halides of molybdenum and tungsten affecting favorably the decyclization. It is assumed that the ring‐opening polymerization of cycloolefins proceeds via π‐allylic complex formation.
Gel formation in cationic polymerization of divinyl ethers. III. Effect of oligooxyethylene chain versus oligomethylene chain as central spacer units
Authors: Hashimoto Tamotsu , Nakamura Tomonori , Tanahashi Sachi , Kodaira Toshiyuki
Published: June 2004

Cationic polymerizations of two series of divinyl ethers were carried out to clarify the effects of their central spacer chain structure on their crosslinking polymerization behavior. One series of the monomers involves divinyl ethers with an oligooxyethylene central spacer chain: diethylene glycol divinyl ether (O‐3), triethylene glycol divinyl ether (O‐4), tetraethylene glycol divinyl ether (O‐5), pentaethylene glycol divinyl ether (O‐6), and heptaethylene glycol divinyl ether (O‐8) (see Scheme 1). The other series includes divinyl ethers with an oligomethylene central spacer chain: 1,4‐butanediol divinyl ether (C‐4), 1,6‐hexanediol divinyl ether (C‐6), and 1,8‐octanediol divinyl ether (C‐8). Cationic polymerizations of these monomers were carried out with the hydrogen chloride/zinc chloride (HCl/ZnCl2) initiating system in methylene chloride (CH2Cl2) at −30 °C ([Monomer]0 = 0.15 M; [HCl]0 = 5.0 mM; [ZnCl2]0 = 0.5 mM). The polymerizations of the oligomethylene‐based divinyl ethers C‐6 and C‐8 caused gel formation at high monomer conversions (∼90%), whereas C‐4 formed soluble polymers even at almost 100% monomer conversion. The oligooxyethylene‐based divinyl ethers O‐3, O‐4, O‐5, and O‐6 underwent gel‐free polymerizations up to 100% monomer conversion and O‐8 did so at least up to ∼80% conversion. The content of unreacted pendant vinyl groups of the obtained soluble polymers was measured by 1H NMR spectroscopy. In the polymerizations of the oligomethylene‐based divinyl ethers (C‐4, C‐6, and C‐8), the vinyl contents of the polymers decreased monotonously with increasing monomer conversion, and their number‐average molecular weights (Mn's) and polydispersity ratios (Mw/Mn's) increased considerably just before the gelation occurred. On the contrary, the vinyl contents of the polymers obtained from the oligooxyethylene‐based divinyl ethers (O‐3, O‐4, O‐5, O‐6, and O‐8) decreased steeply even in the early stage of the polymerizations and almost all the pendant vinyl ether groups were consumed in the soluble polymers at the final stage of the polymerizations. The oligooxyethylene spacer units adjacent to the pendant unreacted vinyl ether groups may solvate intramolecularly with the carbocationic active center to accelerate frequent occurrence of intramolecular crosslinking reactions. © 2004 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 42: 3729–3738, 2004
A model study of circum‐Arctic soil temperatures
Authors: Oelke Christoph , Zhang Tingjun
Published: June 2004

A one‐dimensional heat transfer model with phase change is applied to the entire Arctic terrestrial drainage area. The main forcing parameters are reanalysed surface air temperature that was topography‐enhanced, and snow depth, derived from passive microwave satellite data. We present results based on daily fields of soil temperature, ranging from the soil surface down to 14 m depth. The horizontal grid cell resolution is 25 km × 25 km. Model results are compared to several long series of soil temperature measurements from Alaska and Siberia, with a relatively good match of the the annual cycle at different depths. We show time series of soil temperature at different depths for the 22‐year period 1980 through 2001, and both seasonal and regional analyses are included. A trend analysis reveals positive trends for all permafrost regions in response to positive trends in air temperature, with the strongest warming trend in regions of continuous permafrost. A slight cooling trend is only found for the topmost soil layers in regions of seasonally frozen ground at the southern margins of the Arctic drainage domain. Copyright © 2004 John Wiley & Sons, Ltd.
Circumpolar‐Active‐Layer‐Monitoring (CALM) sites in the Mackenzie Valley, northwestern Canada
Authors: Tarnocai Charles , Mark Nixon F. , Kutny Les
Published: June 2004

Active‐layer studies have been maintained for more than a decade throughout the 1200 km length of the Mackenzie Valley. An important goal of these programmes is to monitor processes linking climate, permafrost and the active layer. Thirteen sites have been selected from these studies for the Circumpolar Active Layer Monitoring programme of the International Permafrost Association. Annual maximum thaw penetration and ground surface movements are measured relative to thaw tubes and soil temperature probes anchored in the permafrost. Active layer thickness, calculated from thaw penetration and surface movement or measured directly, varies more with local soil properties, vegetation and microclimate than with regional atmospheric climate. While depth of thaw penetration has increased at most sites for much of the record, this increase is not always reflected by a proportional increase in active layer thickness because of variable thaw settlement. Air thawing degree‐days (DDT) in the Subarctic and Boreal regions are more than double those in the Arctic regions. These increases in DDT are not reflected in active‐layer depths, suggesting the insulating effect of the surface organic layer, vegetation and snow cover on active‐layer development. Copyright © 2004 John Wiley & Sons, Ltd.
Competing issues for aluminium alloys in structural engineering
Authors: Mazzolani Federico M.
Published: June 2004

This paper makes a contribution to explaining the main reasons for the choice of aluminium alloys as new material in those applications, where they can compete with steel. First of all, the basic physical, mechanical and technological differences from steel are out‐lined in order to identify the main driving issues for justifying a switch from steel to aluminium. Significant results of the application of proper design criteria are shown by examples of many constructions made of aluminium alloys and built around the world in the last few decades in the field of buildings, bridges and special structures.
Intrapartum sonography to determine fetal occipital position: interobserver agreement
Authors: Akmal S. , Tsoi E. , Nicolaides K. H.
Published: June 2004

Objective

To investigate the interobserver agreement on the intrapartum ultrasonographic definition of the fetal occipital position.

Methods

In 60 singleton pregnancies in labor at term the fetal occipital position was determined by transabdominal ultrasound by two appropriately trained sonographers who were not aware of each other's findings. The Bland–Altman plot was performed and the 95% limits of agreement were calculated. Logistic regression analysis was used to investigate the association between complete agreement in the fetal occipital position between the two observers and maternal and labor characteristics.

Results

The two observers had complete agreement on the fetal occipital position in 22/60 (36.7%) cases and disagreement by 15° and 30° in 31 (51.7%) and seven (11.6%) cases, respectively. The mean of the differences between the two observers was 0.25° and the 95% limits of agreement were −28.9° (−32.2° to −25.6°) to 29.4° (26.1° to 32.7°). There were no significant associations between complete agreement and maternal and labor characteristics.

Conclusion

The interobserver agreement on sonographically determined fetal occipital position during labor is within 15° in nearly 90% of cases and within 30° in all cases. Copyright © 2004 ISUOG. Published by John Wiley & Sons, Ltd.
A small heat‐shock protein, p26, from the crustacean Artemia protects mammalian cells (Cos‐1) against oxidative damage
Authors: Collins Crista H. , Clegg James S.
Published: June 2004

A small heat‐shock protein (p26) purified from stress‐resistant embryos of the crustacean, Artemia franciscana, was introduced into cultured cells of green monkey kidney (Cos‐1) using the BioPORTER® delivery system. Cells containing p26 exhibited impressive resistance to hydrogen peroxide compared to controls. Introduction of the disaccharide trehalose did not provide protection against oxidative damage, but enhanced substantially the protective performance of p26 when both were present. These studies extend previous research on the protective role played by p26 in cells exposed to various forms of stress, presumably through its ability to function as a molecular chaperone.
Performance implications of assembly work teams
Authors: McCreery John K. , Krajewski Lee J. , Leong G.Keong , Ward Peter T.
Published: June 2004

This paper explores the role of selected workforce management practices in developing mix, volume, and product flexibility. Using a model of a manually‐paced assembly area, we examine three workforce management practices—the configuration of work teams, the extent of cross training, and the deployment of workers—that have the potential to enhance the level of manufacturing flexibility. We examine the effects of these practices at the level of the individual operation and individual worker, with the goal of maximizing overall system performance in a variety of manufacturing environments. Our results indicate that the value of workforce flexibility is contingent upon characteristics of the operating environment. Environments having high levels of product variety call for the use of a larger number of parallel work teams, while environments with highly complex tasks tend to require a smaller number of parallel teams. Further, the value of cross training and worker task sharing is diminished as work tasks become more complex, due to learning and forgetting effects on the workforce. The overall implication is that more worker flexibility does not always yield improved system performance.
Influence of small imperfections on the stability of plane Poiseuille flow: A theoretical model and direct numerical simulation
Authors: Luo Jisheng , Wu Xuesong
Published: June 2004

It is well known that the stability of plane Poiseuille flow is extremely sensitive to small imperfections that are inevitably present. In this paper, a simple model is proposed, in which the imperfections are represented by a steady but spatially periodic surface roughness and a small oscillatory pressure gradient. A steady perturbation in the form of spatially periodic suction is also considered. For both cases, the resulting steady and unsteady components interact to produce a forcing that is in resonance with the Tollmien–Schlichting (TS) wave. The latter is excited as a result, and grows in proportion to time during the initial stage. The subsequent nonlinear development of the TS wave is shown to be governed by a forced (nonlocal) amplitude equation, which provides a simple framework to link the subcritical nonlinear instability explicitly to external forcing. The validity of the amplitude equation was checked against the direct numerical simulation (DNS), carried out in the case of wall suction. The results indicate that for a fixed level of external disturbance, there exist two distinctive regimes of response: A large-amplitude regime for the Reynolds numbers above a critical value, and a small-amplitude regime below it. The large-amplitude response regime in DNS was found to correspond to the occurrence of a finite-time singularity in the solution to the amplitude equation. This observation allows the critical Reynolds number for the large-amplitude response regime to be defined as one that divides the singular and bounded solutions of the amplitude equation. Estimate based on this equation shows that presence of small imperfections may reduce the critical Reynolds number for subcritical instability to values well below 5772.22.
Small scale structure of homogeneous turbulent shear flow
Authors: Livescu D. , Madnia C. K.
Published: June 2004

The structure of homogeneous turbulent shear flow is studied using data generated by direct numerical simulations (DNS) and a linear analysis for both compressible and incompressible cases. At large values of the mean shear rate, the rapid distortion theory (RDT) limit is approached. Analytical solutions are found for the inviscid compressible RDT equations at long times. The RDT equations are also solved numerically for both inviscid and viscous cases. The RDT solutions, confirmed by the DNS results, show that the even order transverse derivative moments of the dilatational and solenoidal velocity fields are anisotropic, with the dilatational motions more anisotropic than their solenoidal counterparts. The results obtained for the incompressible case are similar to those obtained for the solenoidal motions in the compressible case. The DNS results also indicate an increase in the anisotropy of the even order transverse derivative moments with the order of the moment, in agreement with the RDT predictions. Although the anisotropy decreases with Reynolds number, it is likely that for higher even order moments it will persist at large values of the Reynolds number, in contrast with the postulate of local isotropy. The RDT solutions also predict that the normalized odd order transverse derivative moments of the solenoidal velocity for the compressible case and of the velocity for the incompressible case should approach a constant different than zero at large times. This prediction is supported by the DNS data. For higher odd order normalized moments, the RDT analysis suggests that the anisotropy may persist at large values of the Reynolds number, in agreement with the existent experimental data. The amplification of the dilatational kinetic energy in the direction of the mean shear and the anisotropy of the dilatational dissipation tensor found in the DNS results are also consistent with the RDT analysis.
SUMOylation regulates nucleo-cytoplasmic shuttling of Elk-1
Authors: Salinas Sara , Briançon-Marjollet Anne , Bossis Guillaume , Lopez Marie-Aude , Piechaczyk Marc , Jariel-Encontre Isabelle , Debant Anne , Hipskind Robert A.
Published: June 2004

The transcription factor Elk-1 is a nuclear target of mitogen-activated protein kinases and regulates immediate early gene activation by extracellular signals. We show that Elk-1 is also conjugated to SUMO on either lysines 230, 249, or 254. Mutation of all three sites is necessary to fully block SUMOylation in vitro and in vivo. This Elk-1 mutant, Elk-1(3R), shuttles more rapidly to nuclei of Balb/C cells fused to transfected HeLa cells. Coexpression of SUMO-1 or -2 strongly reduces shuttling by Elk-1 without affecting that of Elk-1(3R), indicating that SUMOylation regulates nuclear retention of Elk-1. Accordingly, overexpression of Elk-1(3R) in PC12 cells, where cytoplasmic relocalization of Elk-1 has been linked to differentiation, enhances neurite extension relative to Elk-1. The effect of Elk-1, but not of the 3R mutant, was blocked upon cotransfection with SUMO-1 or -2 and enhanced by coexpression with mutant Ubc-9. Thus, SUMO conjugation is a novel regulator of Elk-1 function through the control of its nuclear-cytoplasmic shuttling.
Bcl-2–regulated apoptosis and cytochrome c release can occur independently of both caspase-2 and caspase-9
Authors: Marsden Vanessa S. , Ekert Paul G. , Van Delft Mark , Vaux David L. , Adams Jerry M. , Strasser Andreas
Published: June 2004

Apoptosis in response to developmental cues and stress stimuli is mediated by caspases that are regulated by the Bcl-2 protein family. Although caspases 2 and 9 have each been proposed as the apical caspase in that pathway, neither is indispensable for the apoptosis of leukocytes or fibroblasts. To investigate whether these caspases share a redundant role in apoptosis initiation, we generated caspase-2−/−9−/− mice. Their overt phenotype, embryonic brain malformation and perinatal lethality mirrored that of caspase-9−/− mice but were not exacerbated. Analysis of adult mice reconstituted with caspase-2−/−9−/− hematopoietic cells revealed that the absence of both caspases did not influence hematopoietic development. Furthermore, lymphocytes and fibroblasts lacking both remained sensitive to diverse apoptotic stimuli. Dying caspase-2−/−9−/− lymphocytes displayed multiple hallmarks of caspase-dependent apoptosis, including the release of cytochrome c from mitochondria, and their demise was antagonized by several caspase inhibitors. These findings suggest that caspases other than caspases 2 and 9 can promote cytochrome c release and initiate Bcl-2–regulated apoptosis.
Apaf-1 and caspase-9 accelerate apoptosis, but do not determine whether factor-deprived or drug-treated cells die
Authors: Ekert Paul G. , Read Stuart H. , Silke John , Marsden Vanessa S. , Kaufmann Hitto , Hawkins Christine J. , Gerl Robert , Kumar Sharad , Vaux David L.
Published: June 2004

Apoptosis after growth factor withdrawal or drug treatment is associated with mitochondrial cytochrome c release and activation of Apaf-1 and caspase-9. To determine whether loss of Apaf-1, caspase-2, and caspase-9 prevented death of factor-starved cells, allowing them to proliferate when growth factor was returned, we generated IL-3–dependent myeloid lines from gene-deleted mice. Long after growth factor removal, cells lacking Apaf-1, caspase-9 or both caspase-9 and caspase-2 appeared healthy, retained intact plasma membranes, and did not expose phosphatidylserine. However, release of cytochrome c still occurred, and they failed to form clones when IL-3 was restored. Cells lacking caspase-2 alone had no survival advantage. Therefore, Apaf-1, caspase-2, and caspase-9 are not required for programmed cell death of factor-dependent cells, but merely affect its rate. In contrast, transfection with Bcl-2 provided long-term, clonogenic protection, and could act independently of the apoptosome. Unlike expression of Bcl-2, loss of Apaf-1, caspase-2, or caspase-9 would therefore be unlikely to enhance the survival of cancer cells.
The LIM domain protein UNC-95 is required for the assembly of muscle attachment structures and is regulated by the RING finger protein RNF-5 in C. elegans
Authors: Broday Limor , Kolotuev Irina , Didier Christine , Bhoumik Anindita , Podbilewicz Benjamin , Ronai Ze'ev
Published: June 2004

Here, we describe a new muscle LIM domain protein, UNC-95, and identify it as a novel target for the RING finger protein RNF-5 in the Caenorhabditis elegans body wall muscle. unc-95(su33) animals have disorganized muscle actin and myosin-containing filaments as a result of a failure to assemble normal muscle adhesion structures. UNC-95 is active downstream of PAT-3/β-integrin in the assembly pathways of the muscle dense body and M-line attachments, and upstream of DEB-1/vinculin in the dense body assembly pathway. The translational UNC-95::GFP fusion construct is expressed in dense bodies, M-lines, and muscle–muscle cell boundaries as well as in muscle cell bodies. UNC-95 is partially colocalized with RNF-5 in muscle dense bodies and its expression and localization are regulated by RNF-5. rnf-5(RNAi) or a RING domain deleted mutant, rnf-5(tm794), exhibit structural defects of the muscle attachment sites. Together, our data demonstrate that UNC-95 constitutes an essential component of muscle adhesion sites that is regulated by RNF-5.
Nucleocytoplasmic shuttling by nucleoporins Nup153 and Nup214 and CRM1-dependent nuclear export control the subcellular distribution of latent Stat1
Authors: Marg Andreas , Shan Ying , Meyer Thomas , Meissner Torsten , Brandenburg Martin , Vinkemeier Uwe
Published: June 2004

Interferon stimulation of cells leads to the tyrosine phosphorylation of latent Stat1 and subsequent transient accumulation in the nucleus that requires canonical transport factors. However, the mechanisms that control the predominantly cytoplasmic localization in unstimulated cells have not been resolved. We uncovered that constitutive energy- and transport factor-independent nucleocytoplasmic shuttling is a property of unphosphorylated Stat1, Stat3, and Stat5. The NH2- and COOH-terminal Stat domains are generally dispensable, whereas alkylation of a single cysteine residue blocked cytokine-independent nuclear translocation and thus implicated the linker domain into the cycling of Stat1. It is revealed that constitutive nucleocytoplasmic shuttling of Stat1 is mediated by direct interactions with the FG repeat regions of nucleoporin 153 and nucleoporin 214 of the nuclear pore. Concurrent active nuclear export by CRM1 created a nucleocytoplasmic Stat1 concentration gradient that is significantly reduced by the blocking of energy-requiring translocation mechanisms or the specific inactivation of CRM1. Thus, we propose that two independent translocation pathways cooperate to determine the steady-state distribution of Stat1.
Early B Cell Factor Promotes B Lymphopoiesis with Reduced Interleukin 7 Responsiveness in the Absence of E2A
Authors: Seet Christopher S. , Brumbaugh Rachel L. , Kee Barbara L.
Published: June 2004

The basic helix-loop-helix transcription factors encoded by the E2A gene function at the apex of a transcriptional hierarchy involving E2A, early B cell factor (EBF), and Pax5, which is essential for B lymphopoiesis. In committed B lineage progenitors, E2A proteins have also been shown to regulate many lineage-associated genes. Herein, we demonstrate that the block in B lymphopoiesis imposed by the absence of E2A can be overcome by expression of EBF, but not Pax5, indicating that EBF is the essential target of E2A required for development of B lineage progenitors. Our data demonstrate that EBF, in synergy with low levels of alternative E2A-related proteins (E proteins), is sufficient to promote expression of most B lineage genes. Remarkably, however, we find that E2A proteins are required for interleukin 7–dependent proliferation due, in part, to a role for E2A in optimal expression of N-myc. Therefore, high levels of E protein activity are essential for the activation of EBF and N-myc, whereas lower levels of E protein activity, in synergy with other B lineage transcription factors, are sufficient for expression of most B lineage genes.
Cooperative Roles of CTLA-4 and Regulatory T Cells in Tolerance to an Islet Cell Antigen
Authors: Eggena Mark P. , Walker Lucy S.K. , Nagabhushanam Vijaya , Barron Luke , Chodos Anna , Abbas Abul K.
Published: June 2004

Adoptive transfer of ovalbumin (OVA)-specific T cells from the DO.11 TCR transgenic mouse on a Rag−/− background into mice expressing OVA in pancreatic islet cells induces acute insulitis and diabetes only if endogenous lymphocytes, including regulatory T cells, are removed. When wild-type OVA-specific/Rag−/− T cells, which are all CD25−, are transferred into islet antigen–expressing mice, peripheral immunization with OVA in adjuvant is needed to induce diabetes. In contrast, naive CTLA-4−/−/Rag−/− OVA-specific T cells (also CD25−) develop into Th1 effectors and induce disease upon recognition of the self-antigen alone. These results suggest that CTLA-4 functions to increase the activation threshold of autoreactive T cells, because in its absence self-antigen is sufficient to trigger autoimmunity without peripheral immunization. Further, CTLA-4 and regulatory T cells act cooperatively to maintain tolerance, indicating that the function of CTLA-4 is independent of regulatory cells, and deficiency of both is required to induce pathologic immune responses against the islet self-antigen.
Differential Requirement for TANK-binding Kinase-1 in Type I Interferon Responses to Toll-like Receptor Activation and Viral Infection
Authors: Perry Andrea K. , Chow Edward K. , Goodnough Julia B. , Yeh Wen-Chen , Cheng Genhong
Published: June 2004

TANK-binding kinase-1 (TBK1) and the inducible IκB kinase (IKK-i) have been shown recently to activate interferon (IFN) regulatory factor-3 (IRF3), the primary transcription factor regulating induction of type I IFNs. Here, we have compared the role and specificity of TBK1 in the type I IFN response to lipopolysaccharide (LPS), polyI:C, and viral challenge by examining IRF3 nuclear translocation, signal transducer and activator of transcription 1 phosphorylation, and induction of IFN-regulated genes. The LPS and polyI:C-induced IFN responses were abolished and delayed, respectively, in macrophages from mice with a targeted disruption of the TBK1 gene. When challenged with Sendai virus, the IFN response was normal in TBK1−/− macrophages, but defective in TBK1−/− embryonic fibroblasts. Although both TBK1 and IKK-i are expressed in macrophages, only TBK1 but not IKK-i was detected in embryonic fibroblasts by Northern blotting analysis. Furthermore, the IFN response in TBK1−/− embryonic fibroblasts can be restored by reconstitution with wild-type IKK-i but not a mutant IKK-i lacking kinase activity. Thus, our studies suggest that TBK1 plays an important role in the Toll-like receptor–mediated IFN response and is redundant with IKK-i in the response of certain cell types to viral infection.
ON THE SURFACE COMPOSITION OF NORMAL AND SENSITIZED MAMMALIAN BLOOD CELLS
Authors: Mudd Stuart , Mudd Emily B. H.
Published: June 2004

The interfacial tension method has been applied to the study of the surface composition of mammalian blood cells and to certain other particles.

Unsensitized erythrocytes and stromata possess only a small margin of stability in the interface and pass readily into the oil phase.

Specifically sensitized erythrocytes and stromata possess much greater stability in the interface and pass into the oil only with considerable mechanical aid; characteristic deformations of the erythrocyte surface or the interface or both often result.

With special immune sera prepared by Landsteiner and van der Scheer the quantitative relations are such as to indicate that the increased polarity of the sensitized erythrocyte surface is due to combination of the red cell surface lipoids with hemolytic sensitizer. These results are corroborative of the conclusion of Landsteiner and van der Scheer that erythrocytes contain specific lipoid-soluble antigens.

The tentative conclusion is reached that with these anti horse-erythrocyte sera at least the agglutinins combine predominantly with the protein of the red cell surfaces.

Fresh human leucocytes are spread and disintegrated by the interfacial stresses. After heat injury over the condenser with substage lamp the leucocytes typically do not enter the boundary surface. They are pushed before the advancing interface and, if their further advance is obstructed, bend the interface backward to form peninsulas and vacuoles. This change after heating is in the opposite sense to that to be expected from denaturation of the proteins of the protoplasm.

Fresh oxalated rabbit platelets pass very easily into the oils. After heating over the substage lamp these elements also become less oil-miscible.

The interfacial tension relations of blood cells, bacteria, and several cell products are tabulated.
A CHEMICAL STUDY OF THE SPECIFIC ELEMENTS OF TUBERCULIN
Authors: Mueller J. Howard
Published: June 2004

A non-protein gum has been isolated from broth filtrates of human tubercle bacilli grown on a simple medium. It fixes complement and precipitates in high dilution in the presence of homologous immune serum, but fails to give a skin test in tuberculous animals. The preparation and some of the chemical properties of the substance are described.
EXPERIMENTS ON ACTIVE IMMUNIZATION AGAINST EXPERIMENTAL POLIOMYELITIS
Authors: Olitsky Peter K. , Cox Herald R.
Published: June 2004

The results obtained in this investigation indicate that poliomyelitis virus treated with tannin, sodium ricinoleate, or formalin does not constitute a satisfactory immunizing agent in monkeys against the experimental disease.
PATHOLOGY OF PNEUMOCOCCUS INFECTION IN MICE FOLLOWING INTRANASAL INSTILLATION
Authors: Rake Geoffrey
Published: June 2004

Pneumonia can be produced in mice, which have not been previously prepared, by intranasal inoculation of broth cultures of certain strains of pneumococci.

Lesions which are quantitatively different can be produced in different breeds of mice by inoculation of the same type of pneumococcus. Similar inoculation of different types of pneumococci into one breed of mice results in lesions which are qualitatively different.

In general, these lesions are as follows: a diffuse pneumonia and an acute glomerular nephritis in unselected mice receiving Type I strains; a confluent pneumonia and a tubular nephritis in the case of Type II strains; and as result of Type III strains, an interstitial pneumonia with extensive gelatinous pleurisy, together with necrosis and abscess formation in the spleen and cervical lymph nodes. Resistant strains of mice with Type III pneumococci show slight changes in the lungs, but marked lesions in the spleen and cervical nodes, while susceptible mice with the same type of pneumococcus show marked changes in the lung and moderate lesions in the spleen and cervical nodes.

The method of development of Type III pneumonia, studied by means of serial sections of nasally infected mice, appears to proceed in the stages of vascular engorgement, interalveolar interstitial exudate, albuminous fluid exudate into the alveoli and the perivascular lymphatics draining the affected site, and finally, a frank pneumonia with a cellular exudate in the alveoli but without much fibrin.
THE COLONY MORPHOLOGY OF TUBERCLE BACILLI
Authors: Smithburn Kenneth C.
Published: June 2004

The colony topography of tubercle bacilli is significantly affected by altering the pH of the culture medium on which the organisms are grown. Under the conditions of these experiments, avian tubercle bacilli produce two variants, rough and smooth. The former are most numerous on the most acid medium used (pH 6.0); the smooth colonies are obtained over a broad range of pH.

Three colonial variants of bovine and human tubercle bacilli are described. Both mammalian types produce greater numbers of rough colonies at pH 6.0. The bovine type strains produce greatest numbers of smooth colonies in the pH range 6.4 to 6.8, and intermediate colonies on alkaline medium. The human type strains produce greatest numbers of smooth colonies at pH 6.4 and large numbers of intermediate colonies at pH 6.8 and pH 7.2.

Included among the avian and bovine strains studied are organisms of widely varying pathogenic properties. Virulent and attenuated strains of a given type produce similar colonial variants under similar environmental conditions.
A STUDY OF GENERALIZED VACCINIA IN THE CHICK EMBRYO
Authors: Buddingh G. J.
Published: June 2004

1. Chick embryos infected by the chorio-allantoic route with a bacteria-free strain of vaccinia virus develop a general dissemination of the virus throughout the entire organism with the exception of the central nervous system.

2. Quantitative estimation of the distribution of the virus in the various organs of the infected chick by cutaneous inoculation on the rabbit skin offers no evidence for a heightened affinity of the virus for special tissues.

3. Histological study of the lesions in the various organs demonstrates the focal character of the lesions which apparently originate as perivascular infiltrations around the smaller blood vessels. No lesions could be demonstrated in the central nervous system proper.

4. In the earlier stages of the disease Guarnieri bodies are clearly demonstrable in the cells of the epidermis and the squamous epithelium of the buccal mucosa. Inclusion bodies closely resembling Guarnieri bodies are demonstrated in all the lesions occurring in the various other organs.

5. It was not possible to demonstrate conclusively the presence of Paschen bodies in the lesions of the internal organs by the Morosow method usually used for the demonstration of these bodies in the membranal lesion.
BLOOD PLASMA PROTEIN REGENERATION CONTROLLED BY DIET
Authors: McNaught J. B. , Scott V. C. , Woods F. M. , Whipple G. H.
Published: June 2004

When blood plasma proteins are depleted by bleeding, with return of washed red cells (plasmapheresis), it is possible to bring the dog to a steady state of low plasma protein in the circulation and a uniform plasma protein production on a basal diet. These dogs become test subjects by which the potency of various diet factors for plasma protein regeneration can be measured.

Plant and grain proteins are quite well utilized to form new plasma protein in these test dogs but soy bean meal probably should be rated at the head of this list. It is utilized with unexpected promptness and favors the production of albumin in contrast to other plant proteins which distinctly favor globulin production.

Long plasmapheresis periods on basal rations rich in grain proteins lower the resistance of these animals to infection.

Spleen, brain, and stomach when fed with the basal diet in these test dogs show less favorable potency ratios—10.2, 11.8, and 13.6 respectively. This means the grams of tissue protein which must be fed to produce 1 gm. of new plasma protein.

Fasting periods indicate that the dog can contribute only 4 to 6 gm. of plasma protein each week—an insignificant contribution presumably derived from the host's tissue proteins.

Infection and intoxication disturb the plasma protein production of these standardized dogs and may reduce the output of plasma proteins to very low levels in spite of considerable food intake. There may be a very sharp drop in the plasma protein level during the first day of intoxication (Dog 33-324).

Some of these observations may be of value in a study of clinical conditions associated with hypoproteinemia.