bluuebunny/crossref_metadata_2025 huggingface dataset:
Evidence for the molecular heterogeneity of sickle cell anemia chromosomes bearing the βS/Benin haplotype
Authors: Patrinos George P. , Samperi Piera , lo Nigro Luca , Kollia Panagoula , Schiliro Gino , Papadakis Manoussos N.
Published: September 2005

There are at least four distinct African and one Asian chromosomal backgrounds (haplotypes) on which the sickle cell mutation has arisen. Additionally, previous data suggest that the βS/Bantu haplotype is heterogeneous at the molecular level. Here, we report the presence of the Aγ −499 T→A variation in sickle cell anemia chromosomes of Sicilian and North African origin bearing the βS/Benin haplotype. Being absent from North American βS/Benin chromosomes, which were studied previously, this variation is indicative for the molecular heterogeneity of the βS/Benin haplotype. Am. J. Hematol. 80:79–80, 2005. © 2005 Wiley‐Liss, Inc.
Comparison of organ dysfunction in transfused patients with SCD or β thalassemia
Authors: Vichinsky Elliott , Butensky Ellen , Fung Ellen , Hudes Mark , Theil Elizabeth , Ferrell Linda , Williams Roger , Louie Leslie , Lee Phillip D.K. , Harmatz Paul
Published: September 2005

Although it is life saving, transfusion therapy has resulted in the majority of sickle cell anemia and thalassemia patients being at risk for hemosiderosis‐induced organ damage. It is unknown whether the complications of iron overload are affected by the underlying disease. In order to address this problem, we compared the prevalence of organ dysfunction in both groups of patients receiving chronic transfusion therapy (β thalassemia, N = 30; sickle cell anemia, N = 43). Both groups had similar quantitative liver iron. Thalassemia patients had greater cardiac disease (20% vs. 0%), growth failure (27% vs. 9%), and endocrine failure (37% vs. 0%). The strongest predictors of combined endocrine and cardiac disease in multivariate analysis were duration of chronic transfusion (P = 0.03) and diagnosis (P = 0.03). Quantitative liver iron concentration on a single liver biopsy was not predictive of cardiac or endocrine injury. Viral hepatitis is the strongest predictor of hepatocellular damage (P = 0.009), while the development of liver fibrosis is more closely related to liver iron concentration (P = 0.04). In conclusion, sickle cell anemia and thalassemia differ in the prevalence of organ injury. This difference is related to the duration of iron exposure and the specific hemoglobinopathy. A prospective study with a larger number of subjects is needed to confirm the relationships between specific diagnosis, liver iron concentration over time, and organ dysfunction. Am. J. Hematol. 80:70–74, 2005. © 2005 Wiley‐Liss, Inc.
Factor V G1691A (Leiden) and prothrombin G20210A single‐nucleotide polymorphisms in type 2 diabetes mellitus
Authors: Wakim‐Ghorayeb Saria F. , Keleshian Sose H. , Timson Georgina , Finan Ramzi R. , Najm Pierre , Irani‐Hakime Noha , Almawi Wassim Y.
Published: September 2005

The association of the single nucleotide polymorphisms (SNPs) G1691A in coagulation factor V (FV)‐Leiden and G20210A in prothrombin (PRT) genes with type 2 diabetes mellitus (T2DM) were analyzed in 112 T2DM patients (58 males, 54 females; mean age 55.24 ± 13.5 years) and 249 healthy control subjects (118 males, 131 females; mean age 53.03 ± 13.8 years). No association was found for FV‐Leiden with T2DM, as the frequency of the G/G (82.1% vs. 85.5%), G/A (17.0% vs. 14.1%), and A/A (0.9% vs. 0.4%) genotypes was not different between patients and controls, respectively (P = 0.644). Similarly, lack of association of PRT G20210A with T2DM was seen among the population studied, and the frequency of the G/G (92.9% vs. 97.2%), G/A (6.3% vs. 2.8%), and A/A (0.9% vs. 0.0%) genotypes was similar among patients and controls, respectively (P = 0.094). Neither FV‐Leiden nor PRT G20210A was associated with, and no evidence for interactions between these mutations was seen in, T2DM. Am. J. Hematol. 80:84–86, 2005. © 2005 Wiley‐Liss, Inc.
Partial tandem duplication of MLL gene in acute myeloid leukemia with translocation (11;17)(q23;q12–21)
Authors: Yamamoto Shuichi , Nishi Masanori , Taniguchi Kazuto , Imayoshi Miyoko , Ogata Yoshiyasu , Iwanaga Manabu , Sakai Nana , Hamasaki Yuhei , Ishii Eiichi
Published: September 2005

Translocation 11q23 and MLL gene rearrangements are commonly observed in acute myeloid leukemia (AML) in association with the myelomonocytic or monocytic feature. We describe a case involving a 15‐year‐old patient with AML characterized by leukemic cells exhibiting translocation (11;17)(q23;q12–21) and MLL gene rearrangement. No fusion partner gene of the MLL gene was identified, including RARα(17q12) or AF17 (17q21); however, a partial tandem duplication of the MLL exon 11/exon 10 was detected in leukemic cells via a 3′RACE method for detection of unknown partner genes. The patient has been in remission for more than 2 years without hematopoietic stem cell transplantation. Am. J. Hematol. 80:46–49, 2005. © 2005 Wiley‐Liss, Inc.
Rituximab‐induced acute liver failure after an allogeneic transplantation for chronic myeloid leukemia
Authors: Qazilbash Muzaffar H. , Qu Zhenhong , Hosing Chitra , Couriel Daniel , Donato Michele , Giralt Sergio , Champlin Richard
Published: September 2005

Autoimmune hemolytic anemia (AIHA) is a well‐recognized complication of allogeneic bone marrow transplantation (BMT) and can affect up to 5% patients. Several recent case reports suggested the efficacy of anti‐CD20 monoclonal antibody, rituximab, in treating this condition. We report our experience with a 21‐year‐old female with accelerated‐phase chronic myeloid leukemia who underwent allogeneic hematopoietic stem cell transplantation from a matched, unrelated donor. The patient developed autoimmune hemolytic anemia that failed to respond to steroids, intravenous immunoglobulins, and plasma exchange. She was then treated with rituximab that resulted in fatal acute liver toxicity. Am. J. Hematol. 80:43–45, 2005. © 2005 Wiley‐Liss, Inc.
Cancer immunotherapy targeting Sp17: When should the laboratory findings be translated to the clinics?
Authors: Dadabayev Alisher R. , Wang Zhiqing , Zhang Yana , Zhang Jian , Robinson William R. , Lim Seah H.
Published: September 2005

Despite advances in chemotherapeutic agents, the prognosis for some cancers remains extremely poor, suggesting the need for other treatment modalities. Immunotherapy appears an ideal approach because the mechanisms of tumor cell killing induced by tumor vaccines are different from those from chemotherapy. Various investigations are ongoing to identify suitable targets for this purpose. Sperm protein 17 (Sp17) was originally identified by our group as a novel cancer‐testis antigen in various malignancies, including multiple myeloma. Sp17 is a highly immunogenic protein and the observation that more than 90% of vasectomized males develop immunity against Sp17 suggests the opportunity and safety of Sp17 for tumor vaccines. Recent works by other workers suggest a low level of expression of Sp17 in some normal tissues, and investigators have questioned whether Sp17 is in fact a suitable target for immunotherapy. In this paper, we review the general principles of immunotherapy and provide evidence supporting the highly immunogenic nature of Sp17. We also address the discrepancies between the objectives of oncologists involved in treating cancer patients and their familiarity with acceptable levels of toxicity of any effective therapy and those of pure laboratory‐based investigators. Finally, we present some early clinical data supporting the rationale for further investigations of Sp17 for tumor vaccines. Am. J. Hematol. 80:6–11, 2005. © 2005 Wiley‐Liss, Inc.
Acute lymphoblastic leukemia in a patient with chronic granulomatous disease and a novel mutation in CYBB: First report
Authors: Wolach Baruch , Ash Shifra , Gavrieli Ronit , Stark Batia , Yaniv Isaac , Roos Dirk
Published: September 2005

We report for the first time a child with chronic granulomatous disease (CGD) who developed acute lymphoblastic leukemia (ALL). The diagnosis of CGD was made at the age of 4 months, by studies of his neutrophil functions. The superoxide production of the cells was negligible, as was the bactericidal activity. He was found to have a deficiency of the gp91phox subunit of the leukocyte NADPH oxidase, with the X‐linked inheritance of the disease. DNA analysis revealed a C nucleotide insertion between C1028 and T1029. This insertion has not been described before and causes a frameshift and a premature stop codon at amino‐acid position 347. The mother was found to be a carrier of this mutation. At the age of 16 months, the patient developed T‐cell ALL. He was treated for 2 years, and today, 10 years since the diagnosis, he is disease‐free. During the course of ALL and later, he suffered from recurrent severe pyogenic infections, but careful detection of the etiological agent and promptly instituted specific treatment resulted in his complete recovery. Although primary immune deficiencies have been reported to have an increased tendency to develop malignancies, until now there have been no reports of CGD patients with ALL. Am. J. Hematol. 80:50–54, 2005. © 2005 Wiley‐Liss, Inc.
Identification of polymorphisms on the MDR1 gene among Turkish population and their effects on multidrug resistance in acute leukemia patients
Authors: Kaya Pelin , Gündüz Ufuk , Arpaci Fikret , Ural Ali Uĝur , Guran Sefik
Published: September 2005

Multidrug‐resistance (MDR) phenotype is a serious limitation to the effective chemotherapeutic treatment of many cancer types, including leukemia. One of the most important proteins, the over‐expression of which is responsible for the multidrug‐resistance phenotype in many cancer types, is P‐glycoprotein. This protein is the product of the MDR1 gene. In previous studies, single‐nucleotide polymorphisms (SNPs) C3435T, G2677T, and T‐129C in the MDR1 gene were shown to be correlated with lower P‐glycoprotein expression in normal tissues. It was suggested that this might have an advantage in cancer chemotherapy by resulting in a low drug‐resistance phenotype. The frequencies of these SNPs were studied in 45 acute leukemia patients (25 of which were primary refractory and 20 of which were drug‐sensitive) and 17 healthy individuals, forming a Turkish population of 62 individuals. In the first part of the study, these polymorphisms were compared with other populations. Marked differences were apparent between African and Turkish populations for the C3435T polymorphism. On the other hand, similarities were found between other Caucasian/Asian and Turkish populations (P < 0.001). However, for the G2677T polymorphism, the Turkish population is different than Japanese and German populations (P < 0.001). For the T‐129C polymorphism, all individuals in the studied population were homozygous for the T/T genotype. In the second part of this study, drug‐resistant and drug‐sensitive acute leukemia patients were compared for these SNPs. These polymorphisms did not seem to have a significant effect on P‐glycoprotein‐mediated drug resistance in the patients studied. Am. J. Hematol. 80:26–34, 2005. © 2005 Wiley‐Liss, Inc.
Outcome in children with purpura fulminans: Report on 16 patients
Authors: Gürgey Aytemiz , Aytac Selin , Kanra Guler , Secmeer Gulten , Ceyhan Mehmet , Altay Cigdem
Published: September 2005

Purpura fulminans (PF) is a severe disorder of acute onset with high morbidity and mortality. In children, this rapidly progressive illness is usually associated with severe bacterial or viral infections. However, some other conditions may participate in the development of PF. Our objective was to investigate the underlying and associated disorders and the outcomes of the disease in 16 children, 7 males and 9 females ranging in age from 3.5 months to 12 years (median age, 2 years). Thirteen of the 16 children (81%) were 4 years of age or younger. The remaining 3 patients were 9, 10, and 12 years of age. Among these 13 infants and small children, 7 (43%) had infection, 2 infants had congenital cardiac disorders necessitating minor or major surgical intervention, and 1 infant and 3 children had different miscellaneous disorders. The factor V G1691A mutation was present in six of the 13 small children (46%). None of the 3 older children carried the mutation. Six (37.5%) of the 16 patients had protein C deficiencies, and 9 (56%) had protein S deficiencies. These deficiencies, except one for protein S, were acquired. Ten patients except two who were diagnosed at this center were treated with fresh frozen plasma. They were also given heparin. Nine (69%) of the 13 children 4 years of age or younger and one of the older children (33%) required amputation. Five of the six patients (83%) who had factor V G1691A mutation, and who also exhibited severe infection, required amputation. This study suggests that an age of 4 years or less is a risk factor for the development of PF during severe infections, especially in the presence of factor V G1691A mutation and congenital heart disease, necessitating major or minor surgical interventions. This study also shows that the amputation rate in 10 patients, after excluding the patients who had been referred to our center after development of sequelae, was 60%. The survival rate among these 10 patients may indicate that, with the treatment protocol, PF need not be regarded as a lethal disease any more. It is also suggested that effective immunization programs and better health care have probably resulted in some changes in the etiological profile of PF. Am. J. Hematol. 80:20–25, 2005. © 2005 Wiley‐Liss, Inc.
Summer class actions: U.S. and California Supreme Courts revisit arbitrability issues
Authors: Strickland Julia B. , Pearson Scott M. , Newman Stephen J.
Published: September 2005

Julia B. Strickland, Scott M. Pearson and Stephen J. Newman, of Los Angeles, review two summer arbitration rulings, one of which puts arbitrability back in front of the U.S. Supreme Court this fall.
Rule proposal: It's time to provide parties a limited opportunity to seek a rehearing on reasoned arbitration awards
Authors: Gaitis James M.
Published: September 2005

In the second of two parts, James M. Gaitis, of Tucson, Ariz., proposes a rule that would clarify the ability of arbitration parties to correct errors in awards efficiently
Uniform mediation act gets its first high court validation
Authors: Bleemer Russ
Published: September 2005

Details on the New Jersey Supreme Court's analysis using the Uniform Mediation Act, the nation's first high court to address the recently proposed statute.
Die Chemie organischer Nanomaterialien
Authors: Grimsdale Andrew C. , Müllen Klaus
Published: September 2005

Die Entwicklung der Nanotechnologie mit organischen Materialien schreibt eine der größten intellektuellen, aber auch kommerziellen Erfolgsgeschichten unserer Zeit. Fortschritte in der Synthesechemie und bei Methoden zur Untersuchung und Manipulation einzelner Moleküle und kleiner Molekül‐Ensembles brachten entscheidende Fortschritte im Bereich organischer Nanomaterialien hervor. Neue Erkenntnisse über die optischen und elektronischen Eigenschaften von Molekülen, die mit den Methoden der Einzelmolekül‐Spektroskopie und Rastersondenmikroskopie gewonnen wurden, bildeten die Grundlage für den Entwurf und die Synthese neuartiger molekularer und supramolekularer Systeme. Auch für die Anwendung solcher Materialien in optoelektronischen Bauelementen wurde nach Methoden gesucht, wobei Prototypen und Modelle nanoskaliger Funktionseinheiten vielfach beschrieben sind. Ziel dieses Aufsatzes ist es aufzuzeigen, wie das gegenseitige Wechselspiel von Synthesechemie und Spektroskopie das Gebiet der organischen Nanomaterialien hin zum letztendlichen Ziel einer neuen Technologie führt.
The Chemistry of Organic Nanomaterials
Authors: Grimsdale Andrew C. , Müllen Klaus
Published: September 2005

The development of nanotechnology using organic materials is one of the most intellectually and commercially exciting stories of our times. Advances in synthetic chemistry and in methods for the investigation and manipulation of individual molecules and small ensembles of molecules have produced major advances in the field of organic nanomaterials. The new insights into the optical and electronic properties of molecules obtained by means of single‐molecule spectroscopy and scanning probe microscopy have spurred chemists to conceive and make novel molecular and supramolecular designs. Methods have also been sought to exploit the properties of these materials in optoelectronic devices, and prototypes and models for new nanoscale devices have been demonstrated. This Review aims to show how the interaction between synthetic chemistry and spectroscopy has driven the field of organic nanomaterials forward towards the ultimate goal of new technology.
Cover Picture: A T‐Shaped Platinum(II) Boryl Complex as the Precursor to a Platinum Compound with a Base‐Stabilized Borylene Ligand (Angew. Chem. Int. Ed. 35/2005)
Authors: Braunschweig Holger , Radacki Krzysztof , Rais Daniela , Scheschkewitz David
Published: September 2005

Cationic T‐shaped platinum complexes are obtained because of the high trans influence of the ferrocenyl(bromo)boryl ligand. Addition of 4‐methylpyridine to the platinum(II) boryl complex (as shown in the cover picture) causes displacement of the bromide ion at boron, leading to a cationic platinum complex with a 4‐methylpyridine‐stabilized borylene ligand (bottom right). For more information on this approach, see the Communication by H. Braunschweig et al. on page 5651 ff.
Adoption and diffusion of Encoded Archival Description
Authors: Yakel Elizabeth , Kim Jihyun
Published: September 2005

In this article, findings from a study on the diffusion and adoption of Encoded Archival Description (EAD) within the U.S. archival community are reported. Using E. M. Rogers' (1995) theory of the diffusion of innovations as a theoretical framework, the authors surveyed 399 archives and manuscript repositories that sent participants to EAD workshops from 1993–2002. Their findings indicated that EAD diffusion and adoption are complex phenomena. While the diffusion pattern mirrored that of MAchine‐Readable Cataloging (MARC), overall adoption was slow. Only 42% of the survey respondents utilized EAD in their descriptive programs. Critical factors inhibiting adoption include the small staff size of many repositories, the lack of standardization in archival descriptive practices, a multiplicity of existing archival access tools, insufficient institutional infrastructure, and difficulty in maintaining expertise.
Zero valent iron as an electron‐donor for methanogenesis and sulfate reduction in anaerobic sludge
Authors: Karri Srilakshmi , Sierra‐Alvarez Reyes , Field Jim A.
Published: September 2005

Zero valent iron (ZVI) is a reactive media commonly utilized in permeable reactive barriers (PRBs). Sulfate reducing bacteria are being considered for the immobilization of heavy metals in PRBs. The purpose of this study was to evaluate the potential of ZVI as an electron donor for sulfate reduction in natural mixed anaerobic cultures. The ability of methanogens to utilize ZVI as an electron‐donor was also explored since these microorganisms often compete with sulfate reducers for common substrates. Four grades of ZVI of different particle sizes (1.120, 0.149, 0.044, and 0.010 mm diameter) were compared as electron donor in batch bioassays inoculated with anaerobic bioreactor sludge. Methanogenesis was evaluated in mineral media lacking sulfate. Sulfate reduction was evaluated in mineral media containing sulfate and the specific methanogenic inhibitor, 2‐bromoethane sulfonate. ZVI contributed to significant increases in methane production and sulfate reductioncompared to endogenous substrate controls. The rates of methane formation or sulfate reduction were positively correlated with the surface area of ZVI. The highest rates of 0.310 mmol CH4 formed/mol Fe0·day and 0.804 mmol SO reduced/ mol Fe0·day were obtained with the finest grade of ZVI (0.01 mm). The results demonstrate that ZVI is readily utilized as a slow‐release electron donor for methanogenesis and sulfate reduction in anaerobic sludge; and therefore, has a promising potential in bioremediation applications. © 2005 Wiley Periodicals, Inc.
Quantitative methods for developing Fc mutants with extended half‐lives
Authors: Kamei Daniel T. , Lao Bert J. , Ricci Margaret Speed , Deshpande Rohini , Xu Han , Tidor Bruce , Lauffenburger Douglas A.
Published: September 2005

Fc mutants with increased binding affinity for the neonatal receptor, FcRn, exhibit increased half‐lives in vivo, and represent an attractive means for extending the half‐lives of therapeutic antibodies. The half‐lives of other therapeutic molecules (e.g., proteins) may also be extended by conjugating them to Fc fragments, thus decreasing the frequency of patient injections and allowing the administration of low and potentially nontoxic concentrations of the therapeutics. To investigate the possibility for further increasing the half‐life of Fc, a pair of quantitative methods is presented to complement combinatorial screening and in vivo testing. Specifically, a simple molecular modeling procedure was developed to predict relative Gibbs free energies of binding values (ΔΔGbind) between Fc and FcRn across different mutants and species. This procedure was found to reasonably reproduce experimental ΔΔGbind values from our experiments and the literature, and may be used as an initial screen to explore Fc sequence space more fully prior to experimental testing. In addition, a mathematical model of Fc trafficking was formulated and combined with a cell‐level pulse‐chase assay to obtain a quantitative recycling parameter in human T84 cells. This Fc recycling parameter was found to be correlated with binding affinity, but captures the pH dependent nature of the interaction between Fc and FcRn and may serve as an additional screen following combinatorial experiments. © 2005 Wiley Periodicals, Inc.
Fermentation, purification, formulation, and pharmacological evaluation of a prolyl endopeptidase from Myxococcus xanthus: Implications for Celiac Sprue therapy
Authors: Gass Jonathan , Ehren Jennifer , Strohmeier Gregg , Isaacs Indu , Khosla Chaitan
Published: September 2005

Celiac Sprue is a multi‐factorial disease characterized by an inflammatory response to ingested wheat gluten and similar proteins in rye and barley. Proline‐rich gluten peptides from wheat, rye, and barley are relatively resistant to gastrointestinal digestion, and therefore persist in the intestinal lumen to elicit immunopathology in genetically susceptible individuals. In this study, we characterize the in vitro gluten detoxifying properties of a therapeutically promising prolyl endopeptidase from Myxococcus xanthus (MX PEP), and describe the development of a prototypical enteric‐coated capsule containing a pharmacologically useful dose of this enzyme. A high‐cell density fed‐batch fermentation process was developed for overproduction of recombinant MX PEP in E. coli, yielding 0.25–0.4 g/L purified protein. A simple, scalable purification and lyophilization procedure was established that yields >95% pure, highly active and stable enzyme as a dry powder. The dry powder was blended with excipients and encapsulated in a hard gelatin capsule. The resulting capsule was enteric coated using Eudragit L30‐D55 polymer coat, which provided sufficient resistance to gastric conditions (> 1 h in 0.01 M HCl, pH 2 with pepsin) and rapid release under duodenal conditions (15–30 min release in pH 6.0 in the presence of trypsin and chymotrypsin). In conjunction with pancreatic enzymes, MX PEP breaks down whole gluten into a product mixture that is virtually indistinguishable from that generated by the Flavobacterium meningosepticum (FM) PEP as judged by chromatographic assays. Competitive studies involving selected immunogenic peptides mixed with whole gluten reveal that both PEPs have a wide range of substrate specificity. Our results support further in vitro and in vivo evaluation of the MX PEP capsule as an oral therapeutic agent for Celiac Sprue patients. © 2005 Wiley Periodicals, Inc.
Dynamic tradeoffs in the raft‐mediated entry of human immunodeficiency virus type 1 into cells
Authors: Lim Kwang‐il , Yin John
Published: September 2005

To initiate an infection human immunodeficiency virus type 1 (HIV‐1) particles must first bind to receptors on the surface of their host cells, a process that eventually leads to fusion of viral and cellular membranes and release of the viral genome into the cytoplasm. Understanding the molecular mechanisms of these processes may enable the development of new anti‐HIV strategies. Disagreement currently prevails on the role in virus entry of microdomains within the cellular plasma membrane known as lipid rafts. Experiments have suggested that lipid rafts, in their interactions with cellular receptors and viral particles, either promote or have minimal effect on viral entry. Here we develop a dynamic model for HIV‐1 entry that enables us to identify and quantitatively assess tradeoffs that can arise from the clustering of receptors in rafts. Specifically, receptor clustering can be detrimental to the initiation of viral infection by reducing the probability that a virus particle finds its primary receptor, CD4. However, receptor clustering can also enable a virus particle, once bound, to rapidly form multivalent interactions with receptors and co‐receptors that are required for virus‐cell membrane fusion. We show how the resolution of such tradeoffs hinges on the level and spatial distribution of receptors and co‐receptors on the cell surface, and we discuss implications of these effects for the design of therapeutics that inhibit HIV‐1 entry. © 2005 Wiley Periodicals, Inc.
A novel reactive perstraction system based on liquid‐core microcapsules applied to lipase‐catalyzed biotransformations
Authors: Wyss A. , von Stockar U. , Marison I.W.
Published: September 2005

A novel reactive perstraction system has been developed based on liquid‐core capsules, involving an enzyme‐catalyzed reaction coupled with simultaneous in situ product recovery. Lipase‐catalyzed reactions, hydrolysis of triprionin and nitrophenyl laurate, were selected to test the system and demonstrate the feasibility of immobilization of enzymes to the membranes of liquid‐core capsules and the ability to extract hydrophobic products of the reaction within the capsule core. The lipase from Candida rugosa was immobilized to the microcapsules by adsorption and by covalent binding through activation with glutaraldehyde. In both cases improved temperature and operational stability were achieved. Both types of immobilization resulted in a basic shift of the pH optimum for activity, from 7.5 to 9.0. The presence of an organic phase within the capsule core allowed direct product separation and lead to a decrease in product inhibition of the lipase‐catalyzed reaction. © 2005 Wiley Periodicals, Inc.
High‐performance liquid chromatographic assay for the active saponins from Panax notoginseng in rat tissues
Authors: Li Lie , Sheng Yu‐Xin , Zhang Jin‐Lan , Wang Shan‐Shan , Guo De‐An
Published: September 2005

A reversed‐phase liquid chromatographic method was used to determine the ginsenosides Rg1, Rb1 and Rd of Panax notoginseng in rat tissues (kidney, liver, heart, spleen and lung) after the administration of total saponins of P. notoginseng. The tissue samples were treated with solid‐phase extraction prior to HPLC. The calibration curves for the three saponins were linear in the given concentration ranges. The intra‐day and inter‐day assay coefficients in tissues were between 76 and 120% respectively. The recoveries of all the tissues were higher than 70%. This method was applied to evaluate the distribution of the three major saponins of P. notoginseng in rat tissues. Copyright © 2005 John Wiley & Sons, Ltd.
Dynamics‐Based Amplification of RNA Function and Its Characterization by Using NMR Spectroscopy
Authors: Al‐Hashimi Hashim M.
Published: September 2005

The ever‐increasing cellular roles ascribed to RNA raise fundamental questions regarding how a biopolymer composed of only four chemically similar building‐block nucleotides achieves such functional diversity. Here, I discuss how RNA achieves added mechanistic and chemical complexity by undergoing highly controlled conformational changes in response to a variety of cellular signals. I examine pathways for achieving selectivity in these conformational changes that rely to different extents on the structure and dynamics of RNA. Finally, I review solution‐state NMR techniques that can be used to characterize RNA structural dynamics and its relationship to function.
How to Prepare Membrane Proteins for Solid‐State NMR: A Case Study on the α‐Helical Integral Membrane Protein Diacylglycerol Kinase from E. coli
Authors: Lorch Mark , Faham Salem , Kaiser Christoph , Weber Ingrid , Mason A. James , Bowie James U. , Glaubitz Clemens
Published: September 2005

Several studies have demonstrated that it is viable to use microcrystalline preparations of water‐soluble proteins as samples in solid‐state NMR experiments.1–5 Here, we investigate whether this approach holds any potential for studying water‐insoluble systems, namely membrane proteins. For this case study, we have prepared proteoliposomes and small crystals of the α‐helical membrane‐protein diacylglycerol kinase (DGK). Preparations were characterised by 13C‐ and 15N‐cross‐polarization magic‐angle spinning (CPMAS) NMR. It was found that crystalline samples produce better‐resolved spectra than proteoliposomes. This makes them more suitable for structural NMR experiments. However, reconstitution is the method of choice for biophysical studies by solid‐state NMR. In addition, we discuss the identification of lipids bound to membrane‐protein crystals by 31P‐MAS NMR.
Diffusion NMR Spectroscopy: Folding and Aggregation of Domains in p53
Authors: Dehner Alexander , Kessler Horst
Published: September 2005

Protein interactions and aggregation phenomena are probably amongst the most ubiquitous types of interactions in biological systems; they play a key role in many cellular processes. The ability to identify weak intermolecular interactions is a unique feature of NMR spectroscopy. In recent years, pulsed‐field gradient NMR spectroscopy has become a convenient method to study molecular diffusion in solution. Since the diffusion coefficient of a certain molecule under given conditions correlates with its effective molecular weight, size, and shape, it is evident that diffusion can be used to map intermolecular interactions or aggregation events. Complex models can be derived from comparison of experimental diffusion data with those predicted by hydrodynamic simulations. In this review, we will give an introduction to pulsed‐field gradient NMR spectroscopy and the hydrodynamic properties of proteins and peptides. Furthermore, we show examples for applying these techniques to a helical peptide and its hydrophobic oligomerization, as well as to the dimerization behavior and folding of p53.
NMR Methods for Studying the Structure and Dynamics of RNA
Authors: Latham Michael P. , Brown Darin J. , McCallum Scott A. , Pardi Arthur
Published: September 2005

Proper functioning of RNAs requires the formation of complex three‐dimensional structures combined with the ability to rapidly interconvert between multiple functional states. This review covers recent advances in isotope‐labeling strategies and NMR experimental approaches that have promise for facilitating solution structure determinations and dynamics studies of biologically active RNAs. Improved methods for the production of isotopically labeled RNAs combined with new multidimensional heteronuclear NMR experiments make it possible to dramatically reduce spectral crowding and simplify resonance assignments for RNAs. Several novel applications of experiments that directly detect hydrogen‐bonding interactions are discussed. These studies demonstrate how NMR spectroscopy can be used to distinguish between possible secondary structures and identify mechanisms of ligand binding in RNAs. A variety of recently developed methods for measuring base and sugar residual dipolar couplings are described. NMR residual dipolar coupling techniques provide valuable data for determining the long‐range structure and orientation of helical regions in RNAs. A number of studies are also presented where residual dipolar coupling constraints are used to determine the global structure and dynamics of RNAs. NMR relaxation data can be used to probe the dynamics of macromolecules in solution. The power dependence of transverse rotating‐frame relaxation rates was used here to study dynamics in the minimal hammerhead ribozyme. Improved methods for isotopically labeling RNAs combined with new types of structural data obtained from a growing repertoire of NMR experiments are facilitating structural and dynamic studies of larger RNAs.
Solid‐State Magic‐Angle Spinning NMR of Outer‐Membrane Protein G from Escherichia coli
Authors: Hiller Matthias , Krabben Ludwig , Vinothkumar Kutti R. , Castellani Federica , van Rossum Barth‐Jan , Kühlbrandt Werner , Oschkinat Hartmut
Published: September 2005

Uniformly 13C‐,15N‐labelled outer‐membrane protein G (OmpG) from Escherichia coli was expressed for structural studies by solid‐state magic‐angle spinning (MAS) NMR. Inclusion bodies of the recombinant, labelled protein were purified under denaturing conditions and refolded in detergent. OmpG was reconstituted into lipid bilayers and several milligrams of two‐dimensional crystals were obtained. Solid‐state MAS NMR spectra showed signals with an apparent line width of 80–120 Hz (including homonuclear scalar couplings). Signal patterns for several amino acids, including threonines, prolines and serines were resolved and identified in 2D proton‐driven spin‐diffusion (PDSD) spectra.
Structure Refinement of Cyclosporin A in Chloroform by Using RDCs Measured in a Stretched PDMS‐Gel
Authors: Klages Jochen , Neubauer Cajetan , Coles Murray , Kessler Horst , Luy Burkhard
Published: September 2005

New developments concerning alignment media for apolar solvents like chloroform make it possible to measure anisotropic parameters such as residual dipolar couplings (RDCs) at relatively low concentrations and natural isotopic abundance. As RDCs provide structural restraints with respect to an external coordinate system, long‐range structural arrangements of the time‐averaged structure can be determined with high precision. The method is demonstrated on the well‐studied cyclo‐undecapeptide Cyclosporin A (CsA), for which crystal and conventionally derived NMR structures are available. Neither crystal nor NMR structure are consistent with heteronuclear DCH RDCs measured in a stretched poly(dimethylsiloxane) gel, and refinement by using the anisotropic parameter results in a highly defined structure with a slightly changed backbone conformation. The applied methods and interpretation of the structural model are discussed.
Characterisation of Disulfide‐Bond Dynamics in Non‐Native States of Lysozyme and Its Disulfide Deletion Mutants by NMR
Authors: Collins Emily S. , Wirmer Julia , Hirai Kenichi , Tachibana Hideki , Segawa Shin‐ichi , Dobson Christopher M. , Schwalbe Harald
Published: September 2005

This report describes NMR‐spectroscopic investigations of the conformational dynamics of disulfide bonds in hen‐egg‐white lysozyme substitution mutants. The following four systems have been investigated: 2SSα, a lysozyme variant that contains C64A, C76A, C80A and C94A substitutions, was studied in water at pH 2 and 3.8 and in urea (8 M, pH 2); 2SSβ lysozyme, which has C6S, C30A, C115A and C127A substitutions, was studied in water (pH 2) and urea (8 M, pH 2). The NMR analysis of heteronuclear 15N‐relaxation rates shows that the barrier to disulfide‐bond isomerisation can vary substantially in different lysozyme mutants and depends on the residual structure present in these states. The investigations reveal cooperativity in the modulation of micro‐ to millisecond dynamics that is due to the presence of multiple disulfide bridges in lysozyme. Mutation of cysteines in one of the two structural domains substantially diminishes the barrier to rotational isomerisation in the other domain. However, the interactions between hydrophobic clusters within and across the domains remains intact.
Oligonuclear 3d–4f Complexes as Tectons in Designing Supramolecular Solid‐State Architectures: Impact of the Nature of Linkers on the Structural Diversity
Authors: Gheorghe Ruxandra , Cucos Paula , Andruh Marius , Costes Jean‐Pierre , Donnadieu Bruno , Shova Sergiu
Published: September 2005

Heteronuclear cationic complexes, [LCuLn]3+ and [(LCu)2Ln]3+, were employed as nodes in designing high‐nuclearity complexes and coordination polymers with a rich variety of network topologies (L is the dianion of the Schiff base resulting from the 2:1 condensation of 3‐methoxysalycilaldehyde with 1,3‐propanediamine). Two families of linkers have been chosen: the first consists of exo‐dentate ligands bearing nitrogen‐donor atoms (bipyridine (bipy), dicyanamido (dca)), whereas the second consists of exo‐dentate ligands with oxygen‐donor atoms (anions derived from the acetylenedicarboxylic (H2acdca), fumaric (H2fum), trimesic (H3trim), and oxalic (H2ox) acids). The ligands belonging to the first family prefer copper(II) ions, whereas the ligands from the second family interact preferentially with oxophilic rare‐earth cations. The following complexes have been obtained and crystallographically characterized: [LCuII(OH2)GdIII(NO3)3] (1), [{LCuIIGdIII(NO3)3}2(μ‐4,4′‐bipy)] (2), ${{{1\hfill \atop \infty \hfill}}}$[LCuIIGdIII(acdca)1.5(H2O)2]⋅13 H2O (3), ${{{2\hfill \atop \infty \hfill}}}$[LCuIIGdIII(fum)1.5(H2O)2]⋅4 H2O⋅ C2H5OH (4), ${{{1\hfill \atop \infty \hfill}}}$[LCuIISmIII(H2O)(Hfum)(fum)] (5), ${{{1\hfill \atop \infty \hfill}}}$[LCuIIErIII(H2O)2(fum)]NO3⋅3 H2O (6), ${{{2\hfill \atop \infty \hfill}}}$[LCuIISmIII(fum)1.5(H2O)2]⋅4 H2O⋅C2H5OH (7), [{(LCuII)2SmIII}2fum2](OH)2 (8), ${{{1\hfill \atop \infty \hfill}}}$[LCuIIGdIII(trim)(H2O)2]⋅H2O (9), ${{{2\hfill \atop \infty \hfill}}}$[{(LCuII)2PrIII}(C2O4)0.5(dca)]dca⋅2 H2O (10), [LCuIIGdIII(ox)(H2O)3][CrIII(2,2′‐bipy)(ox)2]⋅9 H2O (11), and [LCuGd(H2O)4{Cr(CN)6}]⋅3 H2O (12). Compound 1 is representative of the whole family of binuclear CuII–LnIII complexes which have been used as precursors in constructing heteropolymetallic complexes. The rich variety of the resulting structures is due to several factors: 1) the nature of the donor atoms of the linkers, 2) the preference of the copper(II) ion for nitrogen atoms, 3) the oxophilicity of the lanthanides, 4) the degree of deprotonation of the polycarboxylic acids, 5) the various connectivity modes exhibited by the carboxylato groups, and 6) the stoichiometry of the final products, that is, the CuII/LnIII/linker molar ratio. A unique cluster formed by 24 water molecules was found in crystal 11. In compounds 2, 3, 4, 9, and 11 the CuII–GdIII exchange interaction was found to be ferromagnetic, with J values in the range of 3.53–8.96 cm−1. Compound 12 represents a new example of a polynuclear complex containing three different paramagnetic ions. The intranode CuII–GdIII ferromagnetic interaction is overwhelmed by the antiferromagnetic interactions occurring between the cyanobridged GdIII and CrIII ions.
In situ male breast carcinoma in the Surveillance, Epidemiology, and End Results database of the National Cancer Institute
Authors: Anderson William F. , Devesa Susan S.
Published: September 2005

BACKGROUND

In situ breast carcinoma is not so well characterized for men as for women.

METHODS

Therefore, the authors of the current study compared male and female in situ and invasive breast carcinomas in the Surveillance, Epidemiology, and End Results (SEER) database of the National Cancer Institute to document these patterns.

RESULTS

In situ breast carcinomas composed 9.4% of all male (n = 280 of 2984) and 11.9% of all female breast carcinomas (n = 53,928 of 454,405) during the years 1973–2001. In situ rates rose 123% for men and 555% for women over this time period; whereas distant disease rates fell for both genders. Median ages at diagnosis were 62 years for in situ and 68 years for invasive breast carcinoma among men, compared with 58 years for in situ and 62 years for invasive breast carcinoma among women. Papillary in situ and invasive architectural types were more common among men than women. In contrast, lobular tumors were more common among women than men. Breast cancer‐specific survival was similar among men and women, whereas overall survival was worse for men than women.

CONCLUSION

In situ male breast carcinoma is a rare disease, occurring at older ages and with different architectural types than its more common female counterpart. Gender‐specific histopathologic differences probably reflect anatomic differences among the normal female and vestigial male breast. Rising in situ male breast carcinoma incidence rates over the past three decades suggest earlier detection over time, irrespective of mammography, because men do not participate in routine screening mammography. Worse overall survival for men than women possibly results from age‐dependent comorbid illnesses. Cancer 2005. Published 2005 American Cancer Society.
Merkel cell carcinoma
Authors: McAfee William J. , Morris Christopher G. , Mendenhall Charles M. , Werning John W. , Mendenhall Nancy Price , Mendenhall William M.
Published: September 2005

BACKGROUND

The purpose of the current study was to determine the outcomes of patients with previously untreated Merkel cell carcinoma of the skin who were treated with curative intent.

METHODS

Between October 1984 and January 2002, 34 patients were treated with radiotherapy alone (2 patients) or combined with surgery (32 patients). Nine patients received adjuvant chemotherapy. Patients had follow‐up for a median of 3.0 years (range, 0.3 to 18.5 yrs). Follow‐up on living patients ranged from 2.2 to 18.5 years (median, 7.1 yrs).

RESULTS

The 5‐year outcomes were as follows: local control, 94%, locoregional control, 80%; freedom from distant metastases, 60%; cause‐specific survival, 52%; and survival, 37%. No patient experienced a severe complication.

CONCLUSIONS

Patients treated aggressively with surgery and locoregional radiotherapy have about a 50% chance of cure. Limited data suggest that definitive radiotherapy alone or after incomplete macroscopic resection may control locoregional disease in a significant subset of patients. The dominant site of failure was distant. Cancer 2005. © 2005 American Cancer Society.
Phase II trial of carboplatin and paclitaxel in cisplatin‐pretreated advanced transitional cell carcinoma
Authors: Vaishampayan Ulka N. , Faulkner James R. , Small Eric J. , Redman Bruce G. , Keiser Wayne L. , Petrylak Daniel P. , Crawford E. David
Published: September 2005

BACKGROUND

The purpose of the study was to assess the efficacy and toxicity of carboplatin and paclitaxel administered every 3 weeks in patients with advanced urothelial carcinoma, previously treated with cisplatin‐based therapy.

METHODS

Eligibility included metastatic or locally advanced unresectable transitional cell carcinoma of the urothelial tract. Prior chemotherapy, except taxanes, was permitted within 12 months. Adequate hematologic, hepatic, and renal function and a performance status of 0–2 were required. Treatment consisted of paclitaxel 200 mg/m2 intravenously for 3 hours followed by carboplatin, target area under the curve = 5 repeated every 3 weeks.

RESULTS

Forty‐four patients were enrolled. Thirty‐four (77%) patients had a performance status of 0 or 1. Twenty‐five (57%) of the patients had received prior neoadjuvant or adjuvant chemotherapy, and 19 (43%) had received it for metastatic disease. In all, 181 cycles were administered (median, 3.5 cycles; range, 1–11 cycles). The predominant NCI CTC (version 2.0) Grades 3 and 4 toxicities consisted of myelosuppression in 28 patients and neuropathy in 11 patients. There were no treatment‐related deaths. Of the 44 patients, 1 (2%) had a complete response, 2 (5%) had a partial response, and 4 (9%) had an unconfirmed partial response, for an overall response rate of 16% (95% confidence interval [CI] 7–30%). The median progression‐free survival was 4 months (95% CI 3–5 months) and the median survival was 6 months (95% CI 5–8 months).

CONCLUSIONS

Carboplatin and paclitaxel combination is well tolerated and has modest activity in platinum refractory advanced urothelial carcinoma. Effective regimens need to be developed in cisplatin‐pretreated urothelial carcinoma. Cancer 2005. © 2005 American Cancer Society.
Prevalence of hereditary breast/ovarian carcinoma risk in patients with a personal history of breast or ovarian carcinoma in a mammography population
Authors: Dominguez Francisco J. , Jones Julie L. , Zabicki Katherina , Smith Barbara L. , Gadd Michele A. , Specht Michele , Kopans Daniel B. , Moore Richard H. , Michaelson James S. , Hughes Kevin S.
Published: September 2005

BACKGROUND

Identifying BRCA1 and BRCA2 mutation carriers is increasingly important as new management options show promise in decreasing morbidity and mortality in these women. The authors sought to determine the prevalence of family histories suggestive of a hereditary breast carcinoma syndrome in a cohort of patients with a personal history of breast and/or ovarian carcinoma presenting for mammography.

METHODS

The authors reviewed the family histories of all women with a history of breast or ovarian carcinoma presenting for mammography over a 37‐week period. Using the Myriad model, the authors evaluated the prevalence of family histories with a ≥ 10% risk of a BRCA1 or BRCA2 mutation.

RESULTS

During the period of the current study, 14,597 women completed a family history questionnaire. Of these women, 1764 had a personal history of breast or ovarian carcinoma, 86.6% had unilateral breast carcinoma, 4.6% had bilateral breast carcinoma, 8.2% had ovarian carcinoma, and 0.5% had both breast and ovarian carcinoma. Overall, 20.6% met the criteria for a ≥ 10% risk of mutation according to the Myriad model. This incidence was higher among Ashkenazi women (47.3%) and among patients with a personal history of ovarian carcinoma (35.9%).

CONCLUSIONS

Application of the Myriad model to women with a personal history of breast and ovarian carcinoma suggested that approximately 1 in 5 of these women (20.6%) will have family histories suspicious for a genetic mutation. This risk was higher for Ashkenazi women and for those with a personal history of ovarian carcinoma. This prevalence was considerably higher than the rate reported among women with no personal history of cancer, and has significant implications for their management, as well as for the capacity for risk assessment and testing. Cancer 2005. © 2005 American Cancer Society.
Mature T‐cell leukemias
Authors: Ravandi Farhad , Kantarjian Hagop , Jones Dan , Dearden Claire , Keating Michael , O'Brien Susan
Published: September 2005

Mature T‐cell and NK‐cell leukemias are a group of relatively uncommon neoplasms derived from mature or postthymic T‐cells accounting for a relatively small percentage of lymphoid malignancies. The recent availability of modern immunophenotypic and molecular tools has allowed a better distinction of these disorders from their B‐cell counterparts. Similarly, identification of recurrent cytogenetic abnormalities, as well as plausible mechanisms through which these molecular events influence cellular signaling pathways, have created further insight into the pathogenesis of these disorders. Furthermore, the availability of new agents such as alemtuzumab has generated significant interest in devising specific therapeutic strategies for these malignancies. Herein, we review the clinical and pathological features of mature T‐cell leukemias. Cancer 2005. © 2005 American Cancer Society.