bluuebunny/crossref_metadata_2025 huggingface dataset:
Papillary Necrosis in a Kidney Transplant Following Profound Hypotension
Authors: Raza Muhammad N. , Tomson Charles R.V. , AshMiles Janice
Published: November 2007

We describe a case of a young woman developing profound hypotension requiring inotropic support for 10 days following her second cadaveric renal transplant. Her central venous pressure was adequate, with no evidence of myocardial infarction, and an echocardiogram later showed normal left ventricular function. She had delayed graft function with gradual recovery of her renal functions. On day 45, she developed acute renal failure after removal of a ureteric stent. Ultrasound confirmed hydronephrosis, and a subsequent nephrostogram demonstrated filling defects, likely to be sloughed papillae, causing obstruction. She later had percutaneous removal of necrotic material from the ureter in order to establish free urinary flow with stability of her renal functions. The most likely explanation for papillary necrosis in our patient was profound hypotension postoperatively. We believe our patient's hypotension was secondary to dialysis‐associated cardiovascular autonomic dysfunction, with substantial improvement in blood pressure after a successful renal transplant.
Immunohistochemical expression and distribution of VEGFR‐3 in malignant mesothelioma
Authors: Filho Adhemar Longatto , Baltazar Fátima , Bedrossian Carlos , Michael Claire , Schmitt Fernando C.
Published: November 2007

Homogeneity of mesothelial and lymphatic endothelial cells, express some markers that are presumed to be exclusive of the endothelium was recently reported. This similarity is important to improve the diagnosis and prognosis of malignant mesothelioma (MM). Additionally, some of these markers provide the rationale for specific molecular‐targeted novel therapies aimed at MM, an aggressive malignant neoplasm, with an usually dismal prognosis. The goal of our study was to determine the prevalence and expression pattern of VEGF receptor‐3 (VEGFR‐3) immunoreactivity in MM and whether this immunoreactivity occurs in different phenotypes of this neoplasm. Formalin‐fixed and paraffin‐embedded samples from 29 MM cases and 5 metastatic carcinomas were immuno‐stained for VEGFR‐3 according to the streptavidin–biotin–peroxidase complex technique using a primary antibody (Zymed Laboratories, CA, USA) diluted at 1:200. Lymphatic vessels (LV) were outlined mainly in the peripheral area surrounding the neoplasms. Blood vessels were only rarely positive for VEGFR‐3 in a pattern easily distinguishable from LV. In 25 out of 29 cases (86.2%) LV were strongly positive for VEGFR‐3: 14 cases (48.2%) exhibited positive VEGFR‐3 reactivity in malignant cells. Epitheliod MM showed a moderate to intense VEGFR‐3 positive reaction in LV from 8 out of 19 cases. Among the other histological subtypes, a positive VEGFR‐3 reaction was noted in malignant cells from two cases of transitional and one case of pleomorphic MM. Malignant cells from two out of three biphasic and one out of three sarcomatoid MM were also positive for VEGFR‐3. Interestingly, one case of the multicystic subtype was negative for VEGFR‐3 in malignant cells and faintly positive in an occasional LV. All cases of metastatic carcinoma were negative for VEGFR‐3 in the neoplastic cells. In conclusion, VEGFR‐3 was expressed in malignant cells from different subtypes of MM, reinforcing the putative role of this marker as a potential therapeutic target in this group of neoplasia. Diagn. Cytopathol. 2007;35:786–791. © 2007 Wiley‐Liss, Inc.
MUC4 is upregulated in ovarian carcinoma effusions and differentiates carcinoma cells from mesothelial cells
Authors: Davidson Ben , Baekelandt Mark , Shih Ie‐Ming
Published: November 2007

Using gene expression arrays, we recently showed that MUC4 expression is significantly higher in ovarian/primary peritoneal serous carcinoma (OC/PPC) compared to diffuse peritoneal malignant mesothelioma (DMPM). In the present study, we analyzed the anatomic site‐related expression of MUC4 in OC/PPC and studied its prognostic role. We additionally studied the ability of MUC4 to differentiate between OC/PPC and reactive mesothelial cells (RMC). OC/PPC effusions (n = 142) and benign reactive effusions (n = 10) were immunostained for MUC4 expression. Immunoreactivity was scored in carcinoma cells and RMC and was compared with tumor cell expression in 60 previously studied primary carcinomas and solid metastases and analyzed for association with clinicopathologic parameters, including survival. MUC4 was detected in carcinoma cells in 141/142 (99%) effusions, with comparable expression in peritoneal and pleural effusions. RMC were present in 72 malignant effusions and were MUC4‐negative in all specimens, as well as in the 10 reactive effusions. MUC4 expression in carcinoma cells in effusions was significantly higher than in primary carcinomas and solid metastases (P < 0.001). Higher MUC4 expression was seen in tumors from older (>60 year) patients (P = 0.049). No association was found between MUC4 expression and other clinicopathologic parameters, including survival. MUC4 is universally expressed in OC/PPC effusions and is upregulated at this anatomic site compared to primary carcinomas and solid metastases. The data in the present study, together with our earlier report, show that MUC4 is an excellent marker for differentiating OC/PPC from both benign and malignant mesothelial cells. Diagn. Cytopathol. 2007;35:756–760. © 2007 Wiley‐Liss, Inc.
Blockade of the OX40 ligand prolongs corneal allograft survival
Authors: Hattori Takaaki , Usui Yoshihiko , Okunuki Yoko , Sonoda Yasushi , Usui Masahiko , Takada Eiko , Mizuguchi Junichiro , Yagita Hideo , Okumura Ko , Akiba Hisaya , Takeuchi Masaru
Published: November 2007

Although corneal transplantation is one of the most common tissue transplantations and is known to have a high graft acceptance rate, occasional corneal graft rejection remains a cause of blindness. OX40, a member of the TNF receptor superfamily, is expressed on activated T cells, and transmits a costimulatory signal by binding to OX40 ligand (OX40L) expressed on several cells with antigen‐presenting functions. Using a blocking monoclonal antibody (mAb) against murine OX40L, we investigated the role of OX40 in a murine model of corneal transplantation. C3H/He mouse corneas were transplanted to BALB/c mice orthotopically. Administration of anti‐OX40L mAb significantly reduced allograft rejection, and increased graft survival rate to 40% at 8 weeks after transplantation, while all corneas were rejected within 5 weeks in control IgG‐treated mice. Similar reduced rejection was observed when wild‐type donor corneas were transplanted to OX40L‐deficient recipients. In vitro study revealed that the anti‐OX40L mAb treatment reduced proliferative response and IFN‐γ production of draining lymph node cells in response to stimulation with donor alloantigen. These results demonstrate that OX40L blockade is effective for prolongation of corneal allograft survival by inhibiting recipient T cell activation.
More than Steric Effects: Unlocking the Coordination Chemistry of Barium Pyrazolates
Authors: O'Brien Anna Y. , Hitzbleck Julia , Torvisco Ana , Deacon Glen B. , Ruhlandt‐Senge Karin
Published: November 2007

To further our understanding of the coordination chemistry of the heavy alkaline earth metals, we here report on a family of barium pyrazolates bearing various substitution patterns that illustrate the delicate balance between ligand bulk, donor size, stoichiometry, and hapticity, while emphasizing the importance of secondary interactions including π‐bonding, agostic interactions, and hydrogen bonding to stabilize these heavy‐metal complexes. The dimeric compounds [{Ba(Me2pz)2(pmdta)}2] (1), [{Ba(Phpz)2(pmdta)}2] (2), [{Ba(tBupz)2(pmdta)}2] (3), [{Ba(MePhpz)2(tmeda)}2] (5), [{Ba(Ph2pz)2(tmeda)}2]·TMEDA (6), the polymer [{Ba(tBupz)2(NH3)2}n] (4), and the monomer [Ba(Ph2pz)2(tmeda)2]·TMEDA (7), were prepared using a variety of synthetic methods and were characterized spectroscopically and structurally. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2008)
Use of (S)‐5‐(2‐Methylpyrrolidin‐2‐yl)‐1H‐tetrazole as a Novel and Enantioselective Organocatalyst for the Aldol Reaction
Authors: Tong Sok‐Teng (Amy) , Harris Paul W. R. , Barker David , Brimble Margaret A.
Published: November 2007

The novel organocatalyst (S)‐5‐(2‐methylpyrrolidin‐2‐yl)‐1H‐tetrazole (4) catalyzes the aldol reaction between acetone and various aldehydes with superior enantioselectivity to the existing organocatalysts (S)‐proline (1) and (S)‐5‐(pyrrolidin‐2‐yl)‐1H‐tetrazole (3). (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2008)
Disabled‐1 is a large common fragile site gene, inactivated in multiple cancers
Authors: McAvoy Sarah , Zhu Yu , Perez Damon S. , James C. David , Smith David I.
Published: November 2007

Common fragile sites (CFS) are large, genomically unstable regions, which are hot‐spots for deletions and other alterations, especially in cancer cells. Several have been shown to contain genes that span large genomic regions, such as FHIT (1.5 Mb), WWOX (1.0 Mb), GRID2 (1.36 Mb), PARK2 (1.3 Mb), and RORA (730 kb). These genes are frequently inactivated in multiple different cancers, and FHIT and WWOX are shown to function as tumor suppressors. The disabled‐1 gene (DAB1) is one of the human homologs of the Drosophila disabled locus, which in mammals is involved in neuronal migration and lamination in the developing cerebral cortex. Mice DAB1 inactivation results in the neurological mutant Scrambler, having similarities to mice with the inactivation of PARK2 (Quaker), GRID2 (Lurcher), and RORA (Staggerer). We were interested in whether DAB1 was another large CFS gene that could have cancer development importance. We demonstrated here that the human DAB1 gene (spanning 1.25 Mb) mapped within FRA1B CFS region on chromosomal band 1p32.2. Real‐time RT‐PCR analysis revealed that the expression level of DAB1 was decreased in many human cancer samples, including primary tumor tissues and cancer‐derived cell lines, from several different cancers, especially in brain and endometrial cancer. Additionally, the introduction of an over‐expression DAB1 plasmid into two different cell lines, having insignificant endogenous DAB1 expression, resulted in decreased cell growth. In summary, DAB1 is another gene that resides within an unstable CFS region and might play a role in human tumorigenesis. These data may provide further linkage between neurological development and cancer. © 2007 Wiley‐Liss, Inc.
Sustained complete response and complications rates after radiofrequency ablation of very early hepatocellular carcinoma in cirrhosis: Is resection still the treatment of choice?
Authors: Livraghi Tito , Meloni Franca , Di Stasi Michele , Rolle Emanuela , Solbiati Luigi , Tinelli Carmine , Rossi Sandro
Published: November 2007

If liver transplantation is not feasible, partial resection is considered the treatment of choice for hepatocellular carcinoma (HCC) in patients with cirrhosis. However, in some centers the first-line treatment for small, single, operable HCC is now radiofrequency ablation (RFA). In the current study, 218 patients with single HCC ≤ 2.0 cm (very early or T1 stage) underwent RFA. We assessed 2 primary end points that could be easily compared with those reported for resective surgery: (1) the rate of sustained, local, complete response and (2) the rate of treatment-related complications. The secondary end point was 5-year survival in the 100 patients whose tumors had been considered potentially operable. After a median follow-up of 31 months, sustained complete response was observed in 216 patients (97.2%). In the remaining 6, percutaneous ethanol injection, selective intraarterial chemoembolization, or resection were used as salvage therapy. Perioperative mortality, major complication, and 5-year survival rates were 0%, 1.8%, and 68.5%, respectively.

Conclusion:

Compared with resection, RFA is less invasive and associated with lower complication rate and lower costs. RFA is also just as effective for ensuring local control of stage T1 HCC, and it is associated with similar survival rates (as recently demonstrated by 2 randomized trials). These data indicate that RFA can be considered the treatment of choice for patients with single HCC ≤ 2.0 cm, even when surgical resection is possible. Other approaches can be used as salvage therapy for the few cases in which RFA is unsuccessful or unfeasible. (Hepatology 2007.)
Increased osteoclast formation and activity by peripheral blood mononuclear cells in chronic liver disease patients with osteopenia
Authors: Olivier Brenda J. , Schoenmaker Ton , Mebius Reina E. , Everts Vincent , Mulder Chris J. , van Nieuwkerk Karin M. J. , de Vries Teun J. , van der Merwe Schalk W.
Published: November 2007

Osteoporosis is a common complication of chronic liver disease, and the underlying mechanisms are not understood. We aimed to determine if osteoclasts develop from osteoclast precursors in peripheral blood mononuclear cells (PBMCs) of chronic liver disease patients with osteopenia compared with controls. PBMCs were isolated and fluorescence-activated cell sorting was performed to quantify the activated T lymphocyte population and receptor activator of nuclear factor κβ ligand (RANKL) expression. The activated T lymphocyte populations were comparable for all 3 groups, and RANKL was not detectable. The percentage of CD14+CD11b+ cells containing osteoclast precursors was comparable between the 3 groups. To assess the formation and functional activity of osteoclasts formed from circulating mononuclear cells, PBMCs were cultured (1) without addition of cytokines, (2) with macrophage colony-stimulating factor (M-CSF), (3) with M-CSF and osteoprotegerin, and (4) with M-CSF and RANKL. The number of tartrate-resistant acid phosphatase-positive multinucleated cells and bone resorption was assessed. PBMCs from chronic liver disease patients with osteopenia formed more osteoclast-like cells, which, when cultured in the presence of M-CSF and RANKL resorbed more bone than controls. The number of osteoclast-like cells and the amount of bone resorption correlated with lumbar bone densities. Addition of M-CSF increased numbers of osteoclast-like cells formed in healthy controls; however, this was not observed in either of the chronic liver disease groups. Plasma levels of M-CSF were elevated in both patient groups compared with healthy controls.

Conclusion:

Circulating mononuclear cells from chronic liver disease patients with osteopenia have a higher capacity to become osteoclasts than healthy controls or chronic liver disease patients without osteopenia. This could partially be due to priming with higher levels of M-CSF in the circulation. (Hepatology 2007.)
Nitration Products of 5‐Amino‐1H‐tetrazole and Methyl‐5‐amino‐1H‐tetrazoles – Structures and Properties of Promising Energetic Materials
Authors: Klapötke Thomas M. , Stierstorfer Jörg
Published: November 2007

The nitration of 5‐amino‐1H‐tetrazole (1), 5‐amino‐1‐methyl‐1H‐tetrazole (3), and 5‐amino‐2‐methyl‐2H‐tetrazole (4) with HNO3 (100%) was undertaken, and the corresponding products 5‐(nitrimino)‐1H‐tetrazole (2), 1‐methyl‐5‐(nitrimino)‐1H‐tetrazole (5), and 2‐methyl‐5‐(nitramino)‐2H‐tetrazole (6) were characterized comprehensively using vibrational (IR and Raman) spectroscopy, multinuclear (1H, 13C, 14N, and 15N) NMR spectroscopy, mass spectrometry, and elemental analysis. The molecular structures in the crystalline state were determined by single‐crystal X‐ray diffraction. The thermodynamic properties and thermal behavior were investigated by using differential scanning calorimetry (DSC), and the heats of formation were determined by bomb calorimetric measurements. Compounds 2, 5, and 6 were all found to be endothermic compounds. The thermal decompositions were investigated by gas‐phase IR spectroscopy as well as DSC experiments. The heats of explosion, the detonation pressures, and velocities were calculated with the software EXPLO5, whereby the calculated values are similar to those of common explosives such as TNT and RDX. In addition, the sensitivities were tested by BAM methods (drophammer and friction) and correlated to the calculated electrostatic potentials. The explosion performance of 5 was investigated by Koenen steel sleeve test, whereby a higher explosion power compared to RDX was reached. Finally, the long‐term stabilities at higher temperatures were tested by thermal safety calorimetry (FlexyTSC). X‐Ray crystallography of monoclinic 2 and 6, and orthorhombic 5 was performed.
Alkaloids from the Fruits of Daphniphyllum macropodum
Authors: Wang Xiao‐Ning , Gan Li‐She , Fan Cheng‐Qi , Yin Sheng , Yue Jian‐Min
Published: November 2007

Three new alkaloids, N‐hydroxypaxdaphnine B (1), 21‐O‐acetylpaxdaphnine B (2), and methyl 17‐hydroxyhomodaphniphyllate (3), were isolated from the fresh fruits of Daphniphyllum macropodum, together with six known alkaloids. Their structures were established on the basis of extensive spectroscopic and mass‐spectrometric analyses in combination with chemical transformations.
Alkaloids from the Roots of Stemona cochinchinensis
Authors: Lin Li‐Gen , Dien Pham‐Huu , Tang Chun‐Ping , Ke Chang‐Qiang , Yang Xin‐Zhou , Ye Yang
Published: November 2007

Phytochemical investigation of the roots of Stemona cochinchinensis led to the isolation and structure elucidation of a new pyrido[1,2‐a]azepine‐type alkaloid, stemocochinamine (1), and of four new pyrrolo[1,2‐a]azepine‐type alkaloids, bisdehydrostemocochinine (2), isobisdehydrostemocochinine (3), neostemocochinine (4), and isoneostemocochinine (5), together with six known alkaloids. Their structures were established on the basis of extensive 1D‐ and 2D‐NMR analyses in combination with HR‐MS experiments.
Facile and Selective Synthesis of 4‐Methyl‐ and 4‐Phenylthiosemicarbazide (=N‐Methyl‐ and N‐Phenylhydrazinecarbothioamide) Derivatives of Benzil (=1,2‐Diphenylethane‐1,2‐dione)
Authors: Calatayud David G. , Escolar Francisco J. , López‐Torres Elena , Mendiola M. Antonia
Published: November 2007

A selective synthesis of 4‐methylthiosemicarbazide (=N‐methylhydrazinecarbothioamide; 4a) derivatives by reaction with benzil (=1,2‐diphenylethane‐1,2‐dione; 3) is described. The reaction conditions determined the condensation product formed. The most important factor was the acid used: in the presence of conc. HCl solution, the open‐chain 2 : 1 compound 1a was exclusively obtained, whereas in the presence of 2M HCl, the cyclic 1 : 1 condensation product 2a was formed. The alcohol used, the presence of H2O, and the time of heating were additional crucial factors. The new cyclic compound 2a with a MeO group was exclusively formed when working under high‐dilution conditions. The reaction with the 4‐phenyl derivative 4b gave new cyclic compounds as the major products under all conditions used (Scheme).
A Novel Norlignan and a Novel Phenylpropanoid from Peperomia tetraphylla
Authors: Li Yun‐Zhi , Huang Jing , Gong Zhen , Tian Xiang‐Qing
Published: November 2007

A novel cyclobutane‐type norlignan, peperotetraphin (=methyl rel‐(1R,2S,3S)‐2,3‐bis(7‐methoxy‐1,3‐benzodioxol‐5‐yl)cyclobutanecarboxylate; 1), and a novel phenylpropanoid, i.e., methyl (2E)‐3‐(7‐methoxy‐1,3‐benzodioxol‐5‐yl)prop‐2‐enoate (2), along with three known compounds, α‐asarone (=1,2,4‐trimethoxy‐5‐[(1E)‐prop‐1‐en‐1‐yl]benzene), vanillic acid (=4‐hydroxy‐3‐methoxybenzoic acid), and veratric acid (=3,4‐dimethoxybenzoic acid), were isolated from the EtOH extract of the whole plant of Peperomia tetraphylla. Their structures were determined by spectroscopic methods, especially 1D‐ and 2D‐NMR techniques. This is the first report of naturally occurring cyclobutane‐type norlignans.
Cyclic acetal hydrogel system for bone marrow stromal cell encapsulation and osteodifferentiation
Authors: Betz Martha W. , Modi Parth C. , Caccamese John F. , Coletti Domenick P. , Sauk John J. , Fisher John P.
Published: November 2007

Many systems have been proposed for the encapsulation of bone marrow stromal cells (BMSCs) within degradable hydrogels. Here, we use a novel cyclic acetal‐based biomaterial formed from 5‐ethyl‐5‐(hydroxymethyl)‐β,β‐dimethyl‐1,3‐dioxane‐2‐ethanol diacrylate (EHD) and poly(ethylene glycol) diacrylate (PEGDA). A cyclic acetal‐based hydrogel may be preferred as cyclic acetals hydrolytically degraded into diols and carbonyls as primary degradation products, which may not affect local acidity, unlike other widely investigated polymers. The EHD monomer and PEGDA polymer may be fabricated into a EH‐PEG hydrogel by radical polymerization initiated by the ammonium persulfate (APS) and N,N,N′,N′‐tetramethylethylenediamine (TEMED) system. The objective of this work is to determine whether the components utilized in the fabrication of EH‐PEG hydrogels as well as the EH‐PEG hydrogels permit BMSC viability, metabolic activity, and osteodifferentiation. Cell viability and metabolic activity were assessed after 30 min, 1 h, and 3 h of exposure to pertinent concentrations of the initiator system (10–20 mM). Osteodifferentiation was assessed by alkaline phosphatase and osteocalcin expression after a short exposure to the initiator system to simulate the encapsulation process. Lastly, cell viability was assessed immediately after encapsulation and after 7 days of culture within the EH‐PEG hydrogels. Results indicate that the metabolic activity and viability of BMSCs are minimally affected, and that osteodifferentiation is not significantly affected by the APS‐TEMED initiator system. Also, encapsulated BMSCs maintained viability within EH‐PEG hydrogels for 7 days. This work demonstrates that the EH‐PEG hydrogel is a viable option for the encapsulation and osteodifferentiation of BMSCs. © 2007 Wiley Periodicals, Inc. J Biomed Mater Res 2008
Glutamate accelerates RPE cell proliferation through ERK1/2 activation via distinct receptor‐specific mechanisms
Authors: García Sofía , López Edith , López‐Colomé Ana María
Published: November 2007

The proliferation and migration of Retinal Pigment Epithelium cells resulting from an epithelial‐mesenchymal transition plays a key role in proliferative vitreoretinopathy, which leads to retinal detachment and the loss of vision. In neurons, glutamate has been shown to activate the Ras/Raf/MEK/ERK cascade, which participates in the regulation of proliferation, differentiation, and survival processes. Although glutamate‐stimulation and the activation of ERK1/2 by different stimuli have been shown to promote RPE cell proliferation, the signaling pathway(s) linking these effects has not been established. We analyzed the molecular mechanisms leading to glutamate‐induced proliferation by determining ERK1/2 and CREB phoshporylation in chick RPE cells in primary culture and the human‐derived RPE cell line ARPE‐19. This study shows for the first time, that glutamate promotes RPE cell proliferation by activating two distinct signaling pathways linked to selective glutamate receptor subtypes. Results demonstrate that glutamate stimulates RPE cell proliferation as well as ERK and CREB phosphorylation. These effects were mimicked by the mGluR agonist ACPD and by NMDA, and were prevented by the respective receptor inhibitors MCPG and MK‐801, indicating a cause‐effect relationship between these processes. Whereas mGluR promoted proliferation by activating the MEK/ERK/CREB cascade, NMDA stimulated proliferation through the MEK‐independent activation of Ca2+/calmodulin‐dependent kinases. The blockage of both signaling pathways to proliferation by KN‐62 suggests the involvement of CaMKs in the control of glutamate‐induced proliferation at a common step, downstream of CREB, possibly the regulation of cell cycle progression. Based on these findings, the participation of glutamate in the development of PVR can be considered. J. Cell. Biochem. 104: 377–390, 2008. © 2007 Wiley‐Liss, Inc.
Small tumor antigen of polyomaviruses: Role in viral life cycle and cell transformation
Authors: Khalili Kamel , Sariyer Ilker Kudret , Safak Mahmut
Published: November 2007

The regulatory proteins of polyomaviruses, including small and large T antigens, play important roles, not only in the viral life cycle but also in virus‐induced cell transformation. Unlike many other tumor viruses, the transforming proteins of polyomaviruses have no cellular homologs but rather exert their effects mostly by interacting with cellular proteins that control fundamental processes in the regulation of cell proliferation and the cell cycle. Thus, they have proven to be valuable tools to identify specific signaling pathways involved in tumor progression. Elucidation of these pathways using polyomavirus transforming proteins as tools is critically important in understanding fundamental regulatory mechanisms and hence to develop effective therapeutic strategies against cancer. In this short review, we will focus on the structural and functional features of one polyomavirus transforming protein, that is, the small t‐antigen of the human neurotropic JC virus (JCV) and the simian virus, SV40. J. Cell. Physiol. 215: 309–319, 2008. © 2007 Wiley‐Liss, Inc.
The transient expression of mRNA coding for Rep protein from AAV facilitates targeted plasmid integration
Authors: Howden S. E. , Voullaire L. , Vadolas J.
Published: November 2007

Background

There is a risk of insertional mutagenesis when techniques that facilitate random integration of exogenous DNA into the human genome are used for gene therapy. Wild‐type adeno‐associated virus (AAV) integrates preferentially into a specific site on human chromosome 19 (AAVS1). This is mediated by the interaction of the viral Rep68/78 proteins with Rep‐binding elements in the AAV genome and AAVS1. This specificity is often lost when AAV is used as a gene therapy vector due to removal of the sequences coding for Rep.

Methods

Messenger RNA coding for the Rep68/78 proteins was prepared in vitro and co‐transfected with a 21 kb DNA plasmid containing the P5 integration efficiency element (P5IEE) from AAV. Single cells were seeded in plates to establish clonal cell lines that were subsequently analysed by dual colour fluorescent in situ hybridisation (FISH) to determine whether site‐specific plasmid integration had occurred on chromosome 19.

Results

The co‐transfection of plasmid DNA with Rep68/78 mRNA gave a 2.5‐fold increase in DNA integration when compared to transfection of cells with plasmid DNA alone. Rep68/78 mRNA expression facilitated site‐specific plasmid integration to chromosome 19 in 30% (14/44) of all analysed integration sites, while no targeted integration events were observed following transfection of cells with plasmid DNA alone.

Conclusions

These results demonstrate that transient expression of Rep protein using transfected mRNA facilitates site‐specific integration of plasmid DNA. This approach allows expression of Rep for only a short time, and may circumvent the toxicity and chromosome instability associated with long‐term expression of Rep. Copyright © 2007 John Wiley & Sons, Ltd.
Evaluation of enzymatic treatment of peach juice using response surface methodology
Authors: Santin Márcia M , Treichel Helen , Valduga Eunice , Cabral Lourdes MC , Di Luccio Marco
Published: November 2007

BACKGROUND: This work aimed to assess peach juice clarification using enzymatic treatment. The enzymatic hydrolysis was performed to reduce the juice viscosity and pulp content, and consequently to increase juice extraction. The hydrolysis conditions were optimized by the response surface methodology, where the influence of temperature, amount and type of enzyme and time of reaction were investigated.

RESULTS: Two commercial pectinase preparations (AFP L3 and ULTRA SP WOP from Novozymes) as well as a cellulase (R10 from Onozuka) were tested. The hydrolysis using Pectinex AFP L3 (Novozymes) at 25 °C for 60 min presented the best results of pulp (48%) and viscosity (68%) reduction.

CONCLUSION: Physical and chemical analyses showed that the enzymatic treatment is effective for reduction of peach pulp viscosity, pulp content and turbidity and does not influence other juice parameters such as pH, total acidity, vitamin C and soluble solids. Copyright © 2007 Society of Chemical Industry
Method of fast trace microanalysis of the chiral pesticides epoxiconazole and novaluron in soil samples using off‐line flow‐through extraction and on‐column direct large volume injection in reversed‐phase high performance liquid chromatography
Authors: Rybár Ivan , Góra Robert , Hutta Milan
Published: November 2007

An analytical method combining off‐line flow‐through extraction of a soil micro‐sample (mass around 100 mg, packed into a short HPLC glass column) and direct on‐column large‐volume injection (LVI up to 1.00 mL) of a methanol–water soil extract onto a conventional C18 RP HPLC column enabled fast (within 3.5 minutes) trace micro‐analysis of the relatively new chiral pesticides epoxiconazole (E) and novaluron (N), respectively. Linear calibration curves were evaluated from UV detection (230 nm) data in the range from 0.1 to 5 mg/kg in three most abundant Slovak agricultural soils. LOD (confidence band) at the levels 0.08–0.11 mg/kg and LOQ 0.4–0.6 mg/kg and LOD (S/N = 3) at the levels 0.007–0.018 mg/kg and LOQ (S/N = 10) 0.024–0.060 mg/kg, respectively, of dry soil were achieved. Recovery of pesticides in the overall LVI method including flow‐through 130–200 mg soil micro‐sample extraction was: for epoxiconazole from 74 to 85% and from 56% to 90% for novaluron with reproducibility within ± 6% RSD. This fast (30 min) and simple method consists of just three steps which are short column filling with a solid micro‐sample; flow‐through liquid extraction and direct large‐volume injection RP HPLC DAD analysis. The method is prepared for automation and further analysis of enantiomers of both investigated pesticides by achiral‐chiral column switching techniques.
Comparison between effects of 3‐isobutyl‐1‐methylxanthine and FSH on gap junctional communication, LH‐receptor expression, and meiotic maturation of cumulus–oocyte complexes in pigs
Authors: Ozawa Manabu , Nagai Takashi , Somfai Tamás , Nakai Michiko , Maedomari Naoki , Fahrudin Mokhamad , Karja Ni Wayan Kurniani , Kaneko Hiroyuki , Noguchi Junko , Ohnuma Katsuhiko , Yoshimi Noriko , Miyazaki Hitoshi , Kikuchi Kazuhiro
Published: November 2007

We investigated cAMP content, gap junctional communications (GJCs) status, and LH‐receptor (LH‐R) expression in porcine cumulus–oocyte complexes (COCs) during in vitro maturation treated with the phosphodiesterase (PDE) inhibitor 3‐isobutyl‐1‐methylxanthine (IBMX) or with FSH. COCs were cultured for 20 hr (1st culture) in M199 containing 10% FBS (basic medium, BM group) or BM supplemented with FSH (FSH group) or IBMX (IBMX group). Each COC was then transferred into BM containing both FSH and LH and cultured for an additional 24 hr (2nd culture). The proportions of metaphase‐II (M‐II) oocytes at the end of the 2nd culture did not differ between the FSH (75.7%) and IBMX (68.2%) groups, whereas only 10.1% of oocytes in the BM group reached the M‐II stage. During the 1st culture, the cAMP content of COCs and oocytes became significantly higher in the FSH and IBMX groups than in the BM group; the FSH group had a far greater increment than did the IBMX group. GJCs in the FSH and BM groups gradually closed with increasing duration of the 1st culture, whereas a significantly higher proportion of COCs in the IBMX group still had open GJCs than in the other two groups. Furthermore, LH‐R mRNA expression significantly increased in both the FSH and IBMX groups compared with the BM group. These results suggest that inhibition of PDEs in porcine COCs make the oocyte ready for release from meiotic arrest, and that maintenance of a moderate cAMP content may prolong GJCs and stimulate LH‐R expression. Mol. Reprod. Dev. 75: 857–866, 2008. © 2007 Wiley‐Liss, Inc.
Numerical analysis of block stone displacements in ancient masonry structures: A new method to estimate historic ground motions
Authors: Kamai Ronnie , Hatzor Yossef H.
Published: November 2007

An innovative approach is presented, in which the discontinuous deformation analysis (DDA) method is used to estimate historic ground motions by back analysis of unique structural failures in archaeological sites. Two archaeological sites in Israel are investigated using this new approach and results are presented in terms of displacement evolution of selected structural elements in the studied masonry structure. The response of the structure is studied up to the point of incipient failure, in a mechanism similar to the one observed in the field. Structural response is found to be very sensitive to dynamic parameters of the loading function such as amplitude and frequency.

Prior to back analysis of case studies, two validations are presented. Both compare the performance of DDA with analytical solutions and present strong agreement between the two.

Using comprehensive sensitivity analyses, the most likely peak ground acceleration (PGA) and frequency that must have driven the observed block displacements are found for the two case studies—the Nabatean city of Mamshit and the medieval fortress of Nimrod in southern and northern Israel, respectively. It is found that horizontal peak ground accelerations (HPGA) of 0.5g and 1g were required to generate the observed deformations in Mamshit and Nimrod, respectively. Although these might seem too high, considering structural and topographic amplifications it is concluded that the analyses suggest ground motions of 0.2g at a frequency of 1.5 Hz for Mamshit and up to 0.4g at a frequency of 1 Hz for Nimrod. These values provide constraints on the seismic risk associated with these regions as appears in the local building code using a completely independent approach. Copyright © 2007 John Wiley & Sons, Ltd.
The iterated sinh transformation
Authors: Elliott David , Johnston Peter R.
Published: November 2007

A sinh transformation has recently been proposed to improve the numerical accuracy of evaluating nearly singular integrals using Gauss–Legendre quadrature. It was shown that the transformation could improve the accuracy of evaluating such integrals, which arise in the boundary element method, by several orders of magnitude. Here, this transformation is extended in an iterative fashion to allow the accurate evaluation of similar types of integrals that have more spiked integrands.

Results show that one iteration of this sinh transformation is preferred for nearly weakly singular integrals, whereas two iterations lead to several orders of magnitude improvement in the evaluation of nearly strongly singular integrals. The same observation applies when considering integrals of derivatives of the two‐dimensional boundary element kernel. However, for these integrals, more iterations are required as the distance from the source point to the boundary element decreases. Copyright © 2007 John Wiley & Sons, Ltd.
Stereotyping and nurses' recommendations for treating pain in hospitalized children
Authors: Griffin Ruth A. , Polit Denise F. , Byrne Mary W.
Published: November 2007

The purpose of this study was to examine whether nurses' recommendations for managing children's pain were influenced by stereotypes based on children's personal attributes. Three vignettes, in which hospitalized children's sex, race, and attractiveness were experimentally manipulated, were mailed to a national random sample of 700 pediatric nurses; 334 nurses responded. Responses to vignette questions indicated little evidence of stereotyping. Nurses perceived similar levels of pain and recommended similar pain treatments, regardless of sex, race, and attractiveness. Nurses, on average, perceived children's pain at levels consistent with the children's self‐reports and recommended assertive analgesic and non‐pharmacologic pain management strategies. The results appear consistent with prevailing views on providing adequate pain treatment for children. © 2007 Wiley Periodicals, Inc. Res Nurs Health 30:655–666, 2007
Synthesis of achiral PEG‐PANI rod‐coil block copolymers and their helical superstructures
Authors: Yan Lifeng , Tao Wei
Published: November 2007

Poly(ethylene glycol) (PEG) was modified with aniline groups at both the end, and then PEG‐PANI rod‐coil block polymers have been synthesized by polymerization of the aniline with the aniline‐modified PEG. FTIR, NMR, and elemental analysis provided the chemical strucutre of the as‐prepared polymers. The achiral rod‐coil copolymer could form different superstructures by means of self‐assembly when adding diethyl ether into its THF solution and the length of PANI segments is a key factor to the superstructures. AFM measurements revealed that they form spring‐like helical superstructures from the short PANI‐containing copolymers while these form fibrous helical superstructures from the longer PANI‐containing copolymer. A possible mechanism of the helical superstructures is suggested in this article and the driving force is believed the π–π stacking of the rigid segment of the copolymers. © 2007 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 46: 12–20, 2008
A kind of novel nonmetallocene catalysts for ethylene polymerization
Authors: Ma Lifu , Sheng Yaping , Huang Qigu , Zhao Yangfeng , Deng Kunxue , Li Junlong , Yang Wantai
Published: November 2007

A kind of novel bridged nonmetallocene catalysts was synthesized by the treatment of N,N‐imidazole and N,N‐phenylimidazole with n‐BuLi, and MCl4 (M = Ti, Zr) in THF. Those catalysts were performed for ethylene polymerization after activated by methylaluminoxane (MAO). The effects of polymerization temperature, Al/M ratio, pressure of monomer, and concentration of catalysts on ethylene polymerization behaviors were investigated in detail. Those results revealed that the catalyst system was favorable for ethylene polymerization with high catalytic activity. The polymer was characterized by 13C NMR, WAXD, GPC, and DSC. The result confirmed that the obtained polyethylene featured broad molecular weight distribution around 20, linear structure, and relative low melting temperature. © 2007 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 46: 33–37, 2008
Synthesis of low‐polydispersity poly(N‐ethylmethylacrylamide) by controlled radical polymerizations and their thermal phase transition behavior
Authors: Xu Jian , Jiang Xiaoze , Liu Shiyong
Published: November 2007

Controlled radical polymerizations of N‐ethylmethylacrylamide (EMA) by atom transfer radical polymerization and reversible addition‐fragmentation chain transfer processes were investigated in detail for the first time, employing complementary characterization techniques including gel permeation chromatography, 1H NMR spectroscopy, and matrix‐assisted laser desorption ionization time‐of‐flight mass spectrometry. In both cases, relatively good control of the polymerization of EMA was achieved, as revealed by the linear evolution of molecular weights with monomer conversions and the low polydispersity of poly(N‐ethylmethylacrylamide) (PEMA). The thermal phase transitions of well‐defined PEMA homopolymers with polydispersities less than 1.2 and degrees of polymerization up to 320 in aqueous solution were determined by temperature‐dependent turbidity measurements. The obtained cloud points (CPs) vary in the range of 58–68 °C, exhibiting inverse molecular weight and polymer concentration dependences. Moreover, the presence of a carboxyl group instead of an alkyl one at the PEMA chain end can elevate its CP by ∼3–4 °C. © 2007 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 46: 60–69, 2008
Novel designed polymer–acyclovir conjugates with linker‐controlled drug release and hepatoma cell targeting
Authors: Li Xia , Lu Min , Wu Qi , Lv De‐shui , Lin Xian‐Fu
Published: November 2007

To develop designed polymer–drug conjugates, where the rate of drug liberation and hepatoma cell targeting function could be rationally and widely controlled, we facilely synthesized a series of novel, galactose‐functionalized polymer–acyclovir conjugates with different linkers and first reported the effect of the linker structure including the type of acyclovir‐linked bond (an ester bond or an amide bond) and relative length of the linker between acyclovir and the polymer main chain on release rate and targeting ability of conjugates. In vitro release studies showed that the cumulative released acyclovir from these polymer–acyclovir conjugates was between 24 and 65% in pH 1.2 glycine solution after 7 days. The ester bond more easily underwent hydrolysis than the amide bond. The longer the relative linker length was, the faster the acyclovir was released. The cell recognition experiments visualized using confocal laser scanning microscopy exhibited that the resultant galactose‐functionalized polymer–acyclovir conjugates had evident targeting to hepG2 cells, and targeting ability was also in connection with the relative length of linker. By choosing appropriate linker, cellular internalization of acyclovir could be well achieved. We consider these results to be helpful for the design of multifunctional polymeric prodrugs, in which the required release rate and targeting ability could be rationally controlled by predetermined molecular architecture. © 2007 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 46: 117–126, 2008
Outer‐sphere electron transfer metal‐catalyzed polymerization of styrene using a macrobicyclic ligand
Authors: Bell Craig A. , Whittaker Michael R. , Gahan Lawrence R. , Monteiro Michael J.
Published: November 2007

The CuBr‐catalyzed polymerizations of styrene in the presence of a macrobicyclic mixed donor (N and S) encapsulating ligand, NH2capten, were carried out in toluene at 60 and 100 °C. The macrobicyclic nature of the ligand ensures that a transition metal ion is effectively encapsulated (caged) within the three‐dimensional cavity, resulting in activation of radicals through an outer‐sphere electron transfer mechanism. The kinetic data showed that the polymerizations were uncontrolled with little “living” behavior. The external orders of reaction in [CuBr], [NH2capten], and [CuBr2] were 0.5, 0.5, and close to zero, respectively, in agreement with the postulated mechanism of little or no deactivation of polymeric radicals and a significant amount of bimolecular termination. Although “living” behavior was not found using the cage ligand, it was decided that it would provide an ideal method for radical coupling experiments to make high‐molecular weight multiblock copolymers from a difunctional PSTY (Br‐PSTY‐Br, PDI = 1.11). The coupling reaction of Br‐PSTY‐Br using CuBr/NH2capten and excess Cu(0) in toluene at 100 °C gave no loss of the starting Br‐PSTY‐Br. Changing the solvent to the aprotic DMSO resulted in a significant increase in the rate of consumption of starting Br‐PSTY‐Br, with over 87% being used in under 10 min at 60 °C. In addition, higher molecular weight species were also formed, suggesting that OSET gives little or no side reactions on this time scale. It was initially thought that to get such high rates of reaction that the SET‐LRP disproportionation mechanism (2Cu(I) → Cu(0) + Cu(II)) was at play. However, UV–Vis experiments of the CuBr/NH2capten showed little or no disproportionation products. This important result suggests that DMSO catalyzes the OSET reaction through the stabilization of the radical‐anion intermediate, which then rapidly fragments to a polymeric radical. © 2007 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 46: 146–154, 2008
Proteomics studies of pancreatic cancer
Authors: Chen Ru , Pan Sheng , Aebersold Ruedi , Brentnall Teresa A.
Published: November 2007

Pancreatic cancer is the fourth leading cause of cancer death in the US, only 4% could survive 5 years after diagnosis. Biomarkers are desperately needed to improve earlier, more curable cancer diagnosis and to develop new effective therapeutic targets. The development of quantitative proteomics technologies in recent years offers great promise for understanding the complex molecular events of tumorigenesis at the protein level, and has stimulated great interest in applying the technology for pancreatic cancer studies. Proteomic studies of pancreatic tissues, juice, serum/plasma, and cell lines have recently attempted to identify differentially expressed proteins in pancreatic cancer to dissect the abnormal signaling pathways underlying oncogenesis, and to detect new biomarkers. It can be expected that the continuing evolution of proteomics technology with better resolution and sensitivity will greatly enhance our capability in combating pancreatic cancer.
Overexpression of E‐cadherin and β‐Catenin proteins in metastatic prostate cancer cells in bone
Authors: Saha Baisakhi , Arase Ahren , Imam Sarah S. , Tsao‐Wei Denice , Naritoku Wesley Y. , Groshen Susan , Jones Lawrence W. , Imam S. Ashraf
Published: November 2007

BACKGROUND

The expression of E‐cadherin in the intercellular adhesion of metastatic prostate cancer cells in bone, which is the most prevalent site of metastatic growth, remains elusive.

METHODS

The aim of the study was to compare the concurrent membranous expression of E‐cadherin and β‐catenin proteins, the state which is known to be associated with the cellular adhesion function of E‐cadherin, in prostate biopsy tissue specimens by immunohistochemical staining method. The expression patterns of E‐cadherin or β‐catenin were classified as homogeneous (most cells exhibiting positively), heterogeneous (a few scattered patches of cells with positivity) or negative.

RESULTS

Benign prostate hyperplasia cells exhibited homogeneous expression of both E‐cadherin and β‐catenin in 9 of 11 (82%), whereas the primary prostate cancer cells were homogeneously positive for both proteins only in 4 of 22 (18%) of the cases. The results are similar to those reported in literature. However, in contrast to the primary cancer, a significantly increased frequency of the metastatic prostate cancer cells in bone exhibited homogeneous expression of E‐cadherin and β‐catenin in 12 of 17 (71%) of the cases. A statistically significant association was observed between the overexpression of both proteins and the metastatic prostate cancer cells in bone (Fisher's exact P < 0.001).

CONCLUSIONS

The result of the study demonstrated for the first time that the membranous overexpression of E‐cadherin and β‐catenin are significantly associated with the metastatic prostate cancer cells in bone and that the high frequency of expression suggest their involvement in the intercellular adhesion of the metastatic cells in bone. Prostate 68: 78–84, 2008. © 2007 Wiley‐Liss, Inc.
Neural actions of imidacloprid and their involvement in resistance in the Colorado potato beetle, Leptinotarsa decemlineata (Say)
Authors: Tan Jianguo , Salgado Vincent L , Hollingworth Robert M
Published: November 2007

BACKGROUND: The development of resistance to imidacloprid in eastern US populations of the Colorado potato beetle (CPB), Leptinotarsa decemlineata (Say), threatens this critical use for neonicotinoid insecticides. Previous pharmacokinetic studies with resistant adult CPBs provided no explanation for the high resistance level (over 200‐fold) to topically applied imidacloprid. The authors assessed the neural activity of imidacloprid by recording spontaneous activity from a motor nerve leaving the isolated central nervous system to compare the sensitivity of the latter to imidacloprid between susceptible and resistant CPBs.

RESULTS: On the isolated central nervous system, imidacloprid was initially neuroexcitatory, and neuroinhibitory at higher concentrations. The neuroexcitatory action of imidacloprid was blocked by coapplication of a specific nAChR antagonist, methyllycaconitine, indicating that it is a result of action on nAChRs. The sensitivity to the neuroexcitatory and inhibitory activities of imidacloprid varied independently among individuals in each population. The sensitivity of the central nervous system of resistant CPBs to excitation by imidacloprid did not differ from that of susceptible insects, but the sensitivity to inhibition by imidacloprid was reduced 52‐ to 58‐fold, indicating a possible change in the sensitivity of at least one subgroup of nAChRs.

CONCLUSION: This study provides evidence that reduced nerve sensitivity to the blocking action of imidacloprid is associated with imidacloprid resistance in the CPB. Copyright © 2007 Society of Chemical Industry