bluuebunny/crossref_metadata_2025 huggingface dataset:
Effects of storage time of red blood cell transfusions on the prognosis of coronary artery bypass graft patients
Authors: Van De Watering Leo , Lorinser Jos , Versteegh Michel , Westendord Rudi , Brand Anneke
Published: August 2006

BACKGROUND: In different centers for cardiothoracic surgery throughout the world, different policies are followed concerning the maximum storage time of to‐be‐transfused red blood cells (RBCs). The aim in this study was to investigate the possible role of the storage time of RBC transfusions on the outcome of coronary artery bypass graft (CABG) surgery patients.

STUDY DESIGN AND METHODS: In a single‐center study, all patients who had undergone CABG surgery in the period 1993 until 1999 were identified. Only those patients who had received standard, allogeneic, buffy coat–depleted, unfiltered RBCs in saline‐adenine‐glucose‐mannitol were entered in the analyses (n = 2732). Endpoints were 30‐day survival, hospital stay, and intensive care unit (ICU) stay. Storage time of the perioperative RBC transfusions was analyzed in the following four ways: 1) mean storage time of all perioperative RBC transfusions; 2) storage time of the youngest RBC transfusion; 3) storage time of the oldest RBC transfusion; and 4) comparing outcome in patients receiving only RBCs with a storage time below the median storage of 18 days with patients receiving only RBCs with a storage time above the median.

RESULTS: The univariate analyses showed a strong correlation between storage time and the endpoints survival and ICU stay, but also a correlation with an established risk factor such as the number of transfusions. The multivariate analyses showed no independent effect of storage time on survival or ICU stay.

CONCLUSION: In these analyses, pertaining to 2732 CABG patients, no justification could be found for use of a particular maximum storage time for RBC transfusions in patients undergoing CABG surgery.
A randomized, controlled Phase III trial of therapeutic plasma exchange with fresh‐frozen plasma (FFP) prepared with amotosalen and ultraviolet A light compared to untreated FFP in thrombotic thrombocytopenic purpura
Authors: Mintz Paul D. , Neff Anne , MacKenzie Malcolm , Goodnough Lawrence T. , Hillyer Christopher , Kessler Craig , McCrae Keith , Menitove Jay E. , Skikne Barry S. , Damon Lloyd , Lopez‐Plaza Ileana , Rouault Charles , Crookston Kendall P. , Benjamin Richard J. , George James , Lin Jin‐Sying , Corash Laurence , Conlan Maureen G.
Published: August 2006

BACKGROUND: Photochemical treatment of fresh‐frozen plasma (FFP) with amotosalen and ultraviolet (UV) A light (PCT FFP) results in inactivation of a broad spectrum of pathogens while retaining coagulation factor activity, antithrombotic proteins, and von Willebrand factor–cleaving protease (VWF‐CP) activity.

STUDY DESIGN AND METHODS: A randomized, controlled, double‐blind Phase III trial was conducted with PCT FFP or control FFP for therapeutic plasma exchange (TPE) in patients with thrombotic thrombocytopenic purpura (TTP). Owing to the rarity of this diagnosis, the trial was not powered to demonstrate small differences between treatment groups. Patients were treated with study FFP for a maximum of 35 days until remission was achieved (for a maximum of 30 daily study TPEs with no remission) plus an additional 5 days after remission.

RESULTS: Among the 35 patients treated, the primary endpoint, remission within 30 days, was achieved by 14 of 17 (82%) PCT patients and 16 of 18 (89%) control patients (p = 0.658) The 90 percent confidence interval for treatment difference in remission rate for test − control was (−0.291 to 0.163). Time to remission, relapse rates, time to relapse, total volume and number of FFP units exchanged, and number of study TPEs were not significantly different between groups. Improvement in VWF‐CP and inhibitors was similar for both groups. The overall safety profile of PCT FFP was similar to control FFP. No antibodies to amotosalen neoantigens were detected.

CONCLUSION: The comparable results between treatment groups observed from this small trial suggest that TPE with PCT FFP was safe and effective for treatment of TTP.
Latency‐associated human cytomegalovirus glycoprotein N genotypes in monocytes from healthy blood donors
Authors: Pignatelli Sara , Dal Monte Paola , Rossini Giada , Camozzi Daria , Toscano Vanda , Conte Roberto , Landini Maria Paola
Published: August 2006

BACKGROUND: The β‐herpesvirus human cytomegalovirus (HCMV) infects a variety of cell types and maintains a lifelong relationship with its host by way of a latent infection in circulating monocytes, myeloid precursor cells, and the hematopoietic progenitor population. Viral strain heterogeneity, shown by gene polymorphisms, has been implicated in the majority of HCMV biologic behaviors. HCMV UL73 encodes the polymorphic envelope glycoprotein N (gN), which shows seven genotypes (gN‐1, gN‐2, gN‐3a, gN‐3b, gN‐4a, gN‐4b, and gN‐4c).

STUDY DESIGN AND METHODS: Monocyte subfractions from 64 HCMV‐seropositive healthy blood donors were collected to analyze gN genotypes distribution in the few cells harboring the latent viral genome. Different experimental approaches to extract viral genomes from the monocyte population and amplify UL73 (polymerase chain reaction touchdown and nested) for subsequent genotyping were tested and compared with diagnostic gold standard. gN genotype distribution in monocytes from immunocompetent healthy carriers was compared with previously reported data obtained from patient populations with acute HCMV infections.

RESULTS: The efficiency of UL73 amplification from monocytes of healthy seropositive blood donors was approximately 39 percent, one of the highest reported to date. The leading gN genotype was gN‐1 (87%), whereas the gN‐4 variant was poorly represented (13%). The comparison of gN genotypic frequencies in the immunocompetent healthy population with immunocompromised patients is discussed.

CONCLUSIONS: This work further supports the idea that strain‐specific features could determine the cell tropism and influence the onset of latency.
Surveillance of risk profiles among new and repeat blood donors with transfusion‐transmissible infections from 1995 through 2003 in the Netherlands
Authors: Van der Bij Akke K. , Coutinho Roel A. , Van der Poel Cees L.
Published: August 2006

BACKGROUND: To evaluate the effectiveness of blood donor selection, this study reports risk profiles of donors with transfusion‐transmissible infections as obtained by ongoing surveillance, 1995 through 2003, in the Netherlands.

STUDY DESIGN AND METHODS: A surveillance program was installed to monitor risk profiles among new and repeat donors infected with human immunodeficiency virus (HIV), hepatitis C virus (HCV), hepatitis B virus (HBV), or human T‐lymphotropic virus (HTLV), or positive for the presence of syphilis antibodies. At posttest counseling, a physician interviewed donors to clarify possible sources of infection.

RESULTS: A total of 167 repeat donors and 404 new donors were interviewed: 33 with HIV, 123 with HCV, 279 with HBV, 21 with HTLV, and 112 with syphilis antibodies. Most HBV, HCV, and HTLV infections were among new donors (80, 85, and 67%), whereas most HIV infections were among repeat donors (79%). Nearly 25 percent of the donors did not report factors at screening that would have deferred them from donating blood. At posttest interviews, new donors with HCV often reported injecting drug use (19%). Repeat donors with HIV often reported male‐to‐male sex (8/26, 31%).

CONCLUSION: A significant level of deferrable behavioral risks was found among donors with confirmed transfusion‐transmissible infections that persist despite current donor selection. Reporting such behavior at initial donor selection would have eliminated a substantial part of the infections found. This study argues against relaxing the existing donor deferral of persons practicing male‐to‐male sex, given their significant proportion of HIV infections among repeat donors. Systematic surveillance of risk factors among infected blood donors provides ongoing information about the effectivity of donor selection and is recommended to evaluate and optimize blood policies.
Molecular basis of the LOCR (Rh55) antigen
Authors: Coghlan Gail , Moulds Marilyn , Nylen Edward , Zelinski Teresa
Published: August 2006

BACKGROUND: In 1994 during the investigation of a case of hemolytic disease of the newborn, a new low‐incidence red cell (RBC) antigen, LOCR, was described. Although the presence of LOCR was associated with altered expression of Rh antigens, its formal assignment to the Rh blood group system did not occur until haplotype and linkage analysis conducted in 2003 provided the necessary proof. The current study was undertaken in an attempt to define the underlying RH mutation in LOCR+ individuals.

STUDY DESIGN AND METHODS: Genomic DNA from five unrelated LOCR+ individuals and three Rh‐matched control individuals was amplified by polymerase chain reaction with intronic primers flanking all 10 exons of RH. Amplified products were separated on 1 percent agarose gels and isolated for DNA sequence analysis in both the forward and the reverse directions.

RESULTS: DNA sequence analysis of the three LOCR+ D– individuals revealed a single heterozygous 286G>A nucleotide substitution resulting in a predicted Gly>Ser substitution at amino acid 96. DNA sequence analysis from the two LOCR+ D+ individuals revealed the identical mutation, as well as all of the changes associated with the common RHD gene.

CONCLUSIONS: Based on our results, a Gly96Ser substitution in the Rhce polypeptide defines the low‐incidence RBC antigen known as LOCR. This same amino acid change has previously been shown to be involved in the Rh:‐26 phenotype, which suggests that LOCR and Rh26 are antithetical. Serologic investigations with various Rh:‐26 cells and serum samples, however, reveal that only some c+ Rh:‐26 phenotypes are LOCR+.
Viability and function of 8‐day‐stored apheresis platelets
Authors: Slichter Sherrill J. , Bolgiano Doug , Jones Mary Kay , Christoffel Todd , Corson Jill , Rose Leslie , Foley Jim , Popovsky Mark , Baril Laurene L. , Corda Tammy , Dincecco Dorothy M. , Snyder Edward L.
Published: August 2006

BACKGROUND:Methods of bacterial detection and pathogen inactivation of platelets (PLTs) may allow extended storage of PLTs as long as PLT quality is maintained.

STUDY DESIGN AND METHODS:Twenty normal volunteers had their PLTs collected with an apheresis machine (Haemonetics Corp.). A variety of in vitro PLT function and metabolic assays were performed both on Day 0 and after 8 days of storage. On Day 8, a small blood sample was drawn from each donor to obtain fresh PLTs. The fresh and stored autologous PLTs were labeled with either51Cr or111In, and the radiolabeled PLTs were transfused. Posttransfusion serial blood samples were drawn to determine the relative posttransfusion recoveries and survivals of the fresh versus the stored PLTs.

RESULTS:Although the in vitro assays showed some differences between the two trial sites, the results were generally within the ranges expected for fresh and stored PLTs. Overall, PLT recoveries averaged 66 ± 16 percent versus 53 ± 20 percent and survivals averaged 8.5 ± 1.6 days versus 5.6 ± 1.6 days, respectively, for fresh compared to 8‐day‐stored PLTs. There were no significant differences in the in vivo PLT data between the trial sites or based on the radiolabel used for the measurements.

CONCLUSION:After 8 days of storage, the in vivo posttransfusion recovery and survival of autologous Haemonetics apheresis PLTs meet the proposed standards for poststorage PLT quality.
Embedded Localities: Employment Decline, Inner City Population Growth, and Declining Place Stratification among Japan's Mid‐Size and Large Cities
Authors: Jacobs A. J.
Published: August 2006

The 1990s were a decade of economic decline in Japan. During this period, similar to the United States in the 1970s and 1980s, Japan's largest cities suffered contractions in total and manufacturing employment, and income inequality widened, nationwide. In contrast, after 1995, inner city districts in eight of its 11 largest cities and in central Tokyo experienced population and housing growth. Residential growth, coupled with contracting incomes in affluent cities and rising incomes in provincial large and mid‐sized cities, then fostered a decline in place stratification by per capita and household income among cities with more than 200,000, between 1990 and 2000. This article argues that the primary factors provoking this combination of positive and negative outcomes were the same policies implemented by the Japanese Developmental State in order to protect its firms from the destructive forces of the world capitalist market system. In other words, failed market interventions unintentionally led to industrial disinvestment and plunging urban land prices. These conditions, supported by national and subnational urban redevelopment policies, then spurred residential growth in large cities. In sum, Japan's interventionist policy logic and its unique development trends suggest that its cities remain as embedded localities, tightly nested within their own political‐economic development context. Its situation also suggests that it might be time to create a more integrated metatheory of urban development, one which fully incorporates the reflexive nature of the interplay among the global, the national, and the local political‐economic contexts in order to explain its impacts on spatial patterns.
Private, Parochial, and Public Realms in Santiago, Chile's Retail Sector
Authors: Stillerman Joel
Published: August 2006

Recent scholarship contends that the rise of shopping malls, gated communities, and gentrification as well as citizens' withdrawal to the private realm have eroded public life in U.S. and Latin American cities. Malls' suburban location and security policies exclude the poor and restrict free speech; residents and fences in gated communities exclude outsiders; and police and businesses in downtowns and high‐rent districts limit poor people's access to public areas. I expand this discussion with an analysis of the accessibility of Santiago, Chile's retail areas, the social relationships present there, and marginalized groups' informal resistance to their exclusion. The city's distinct segregation pattern, transit system, and state‐licensed street markets permit greater contact between rich and poor and foster vital public spaces. I adapt Lofland's typology of fleeting, quasi‐primary, and intimate secondary relations in public to examine social interactions in street markets, flea markets, and shopping malls. The distinct mix of relationships within these markets reflects the characteristics of users, varying degrees of accessibility to diverse populations, and state policies toward markets. Marginalized groups' informal resistance is pervasive in each setting. In contrast to the dominant view that public space is declining in contemporary cities, Santiago residents are not universally reclusive, antisocial, or reluctant to engage in cross‐class public encounters, and the city retains vital public areas. The findings demonstrate that our understanding of public space is incomplete without an awareness of social relationships and informal resistance alongside structural constraints to the accessibility of urban locales.
Predictors of long‐term work disability in Major Depressive Disorder: a prospective study
Authors: Rytsälä H. J. , Melartin T. K. , Leskelä U. S. , Sokero T. P. , Lestelä‐Mielonen P. S. , Isometsä E. T.
Published: August 2006

Objective:  Major Depressive Disorder (MDD) is a major cause of long‐term work disability. However, factors predicting this are not well known.

Method:  In the Vantaa Depression Study, predictors for being granted a disability pension during an 18‐month follow‐up were examined among the 186 psychiatric MDD in‐ and out‐patients belonging to the labour force at baseline.

Results:  The 21 patients (11.3%) granted a disability pension were significantly older, more hopeless, had worse social and occupational functioning, and spent more time depressed during follow‐up. After adjusting for these predictors, being on sick leave at baseline still strongly predicted disability pension during follow‐up.

Conclusion:  Disability pension is predicted by multiple sociodemographic and clinical factors. Baseline level of functioning and duration of depressive episodes are key clinical predictors. The positive and negative consequences of sick leave warrant closer attention.
Relapse rates in an early psychosis treatment service
Authors: Addington D. , Jean Addington M. D. , Patten S.
Published: August 2006

Objective:  To examine the relapse rate achievable in a real‐life early psychosis treatment service.

Method:  A 2‐year longitudinal cohort study of consecutive admissions to an early psychosis programme which served the entire population in a catchment area. The primary outcome measure was relapse.

Results:  One hundred and forty‐eight consecutive consenting admissions were recruited, 124 (83.8%) of whom were followed for 1 year and 116 (78.4%) for 2 years. Relapse was assessed by clinicians using structured criteria. The 2‐year relapse rate among subjects with complete data collection was 34/95 (35.7%, 95% CI 26.2–46.3). A Kaplan–Meier life table censoring subjects lost to follow‐up yielded a comparable estimate of the proportion not relapsing: 68% (95% CI 58–76%).

Conclusion:  These estimates compare favourably with a published range of 2‐year relapse from 55% to 70% in the older literature and are comparable with the results in recent clinical trials.
Preclinical evaluation of pharmacokinetics and safety of melatonin in propylene glycol for intravenous administration
Authors: Cheung Raymond Tak Fai , Tipoe George L. , Tam Sidney , Ma Edmond Shiu Kwan , Zou Liang Yu , Chan Pui Shan
Published: August 2006

Abstract:  Melatonin is a highly effective treatment in different animal models of excitotoxicity or ischemia/reperfusion injury. Due to a lack of patentability, commercial sponsors are not interested in funding clinical evaluations of melatonin. Investigators may initiate small‐scale clinical evaluation, and intravenous (i.v.) administration is appropriate in acute stroke patients. Institutional Review Boards may require proper preclinical evaluation of the preparation. In this pharmacokinetic and safety study, melatonin in propylene glycol was evaluated in adult male Sprague–Dawley rats. Following a single i.v. injection at 5 or 15 mg/kg, plasma concentrations of melatonin increased to 39 and 199 million pg/mL at 2 min and 128 000 and 772 000 pg/mL at 120 min. Within 60 min of injection, the blood pressure, heart rate and body temperature remained unaffected. Melatonin at 5 mg/kg did not influence the complete blood counts at 60 min, but melatonin at 15 mg/kg had some effects on the differential white cell and platelet counts. Melatonin at 5 or 15 mg/kg slightly elevated some liver enzymes at 60 min of injection, and melatonin at higher dose also elevated plasma creatinine and lactate dehydrogenase levels. At 24 hr after completion of six daily injections of melatonin, there was a 5.5% reduction in body weight. Gross postmortem examination and histological examination of the brain, kidney, liver and spleen did not reveal any evidence of toxicity. In conclusion, melatonin in propylene glycol markedly elevates plasma levels of melatonin with no serious toxicity. This preparation should be further evaluated in human patients.
Birds as Biodiversity Indicators for Europe
Authors: Gregory Richard
Published: August 2006

It is a truism that biodiversity is vital. It is, after all, a measure of life on earth. It is of more pressing concern that biodiversity is being lost at a rate unprecedented in the recent era, and will continue to be lost unless action is taken. There is species loss and habitat loss across ecosystems from the Arctic icecap to tropical rainforest: the so-called Fifth Great Extinction of species really does seem to be happening. Richard Gregory looks at the role birds can play in measuring biodiversity.
Safety Cameras: Stealth Tax or Life-Savers?
Authors: Mountain Linda
Published: August 2006

There cannot be a road user in the UK who does not have an opinion on speed cameras—in most cases a rather strongly held opinion. Indeed many of us will hold them responsible for the first blemish on an otherwise clean license. Their proper name is not speed but safety cameras—but do they really save lives, or are they just one more way of extracting money from the hapless motorist? Linda Mountain knows the answers.
A Case of Mistaken Identity
Authors: Davies Simon
Published: August 2006

The UK government has embarked on the world's most ambitious national identity project. During its 3-year progress towards Royal Assent the scheme has variously been proclaimed as a key solution to identity theft, illegal immigration, administrative inefficiency, benefit fraud, VAT fraud, illegal working, terrorism, healthcare tourism, national security, telecommunications fraud and organized crime. Can it actually work? Simon Davies has his doubts.
Statistics and The Bayesian Mind
Authors: Griffiths Thomas L.
Published: August 2006

Did German flying bombs really target the poorer areas of London—or did the poor people underneath them just think that they did? How many millions of dollars will the current re-make of King Kong really gross? Oh, and how many taxis are there in London? The human mind can make very good guesses, it seems—and it uses Bayesian analysis to do so. Tom Griffiths and Joshua Tenenbaum explain.
Trading Coupons: Completing the World Cup Football Sticker Album
Authors: Hayes Kevin , Hannigan Ailish
Published: August 2006

The agonies and the ecstasies of the World Cup are over for another four years. England supporters may not wish to be reminded of Ronaldo's spot-kick, and we are loath to add to pain. But one set of players who probably did very well from the competition was Panini, the company which produced the official 2006 World Cup football sticker album—and, more importantly, the collectable stickers which go inside it. Kevin Hayes and Ailish Hannigan tried to fill up their albums.
Fatty acid regulation of adenylyl cyclase Rv2212 from Mycobacterium tuberculosis H37Rv
Authors: Motaal Amira Abdel , Tews Ivo , Schultz Joachim E. , Linder Jürgen U.
Published: August 2006

Adenylyl cyclase Rv2212 from Mycobacterium tuberculosis has a domain composition identical to the pH‐sensing isoform Rv1264, an N‐terminal regulatory domain and a C‐terminal catalytic domain. The maximal velocity of Rv2212 was the highest of all 10 mycobacterial cyclases investigated to date (3.9 µmol cAMP·mg−1·min−1), whereas ATP substrate affinity was low (SC50 = 2.1 mm ATP). Guanylyl cyclase side activity was absent. The activities and kinetics of the holoenzyme and of the catalytic domain alone were similar, i.e. in distinct contrast to the Rv1264 adenylyl cyclase, in which the N‐terminal domain is autoinhibitory. Unsaturated fatty acids strongly stimulated Rv2212 activity by increasing substrate affinity. In addition, fatty acids greatly enhanced the pH sensitivity of the holoenzyme, thus converting Rv2212 to a pH sensor adenylyl cyclase. Fatty acid binding to Rv2212 was modelled by homology to a recent structure of the N‐terminal domain of Rv1264, in which a fatty acid‐binding pocket is defined. Rv2212 appears to integrate three cellular parameters: ATP concentration, presence of unsaturated fatty acids, and pH. These regulatory properties open the possibility that novel modes of cAMP‐mediated signal transduction exist in the pathogen.
Association of mammalian sterile twenty kinases, Mst1 and Mst2, with hSalvador via C‐terminal coiled‐coil domains, leads to its stabilization and phosphorylation
Authors: Callus Bernard A. , Verhagen Anne M. , Vaux David L.
Published: August 2006

Genetic screens in Drosophila have revealed that the serine/threonine kinase Hippo (Hpo) and the scaffold protein Salvador participate in a pathway that controls cell proliferation and apoptosis. Hpo most closely resembles the pro‐apoptotic mammalian sterile20 kinases 1 and 2 (Mst1 and 2), and Salvador (Sav) has a human orthologue hSav (also called hWW45). Here we show that Mst and hSav heterodimerize in an interaction requiring the conserved C‐terminal coiled‐coil domains of both proteins. hSav was also able to homodimerize, but this did not require its coiled‐coil domain. Coexpression of Mst and hSav led to phosphorylation of hSav and also increased its abundance. In vitro phosphorylation experiments indicate that the phosphorylation of Sav by Mst is direct. The stabilizing effect of Mst was much greater on N‐terminally truncated hSav mutants, as long as they retained the ability to bind Mst. Mst mutants that lacked the C‐terminal coiled‐coil domain and were unable to bind to hSav, also failed to stabilize or phosphorylate hSav, whereas catalytically inactive Mst mutants that retained the ability to bind to hSav were still able to increase its abundance, although they were no longer able to phosphorylate hSav. Together these results show that hSav can bind to, and be phosphorylated by, Mst, and that the stabilizing effect of Mst on hSav requires its interaction with hSav but is probably not due to phosphorylation of hSav by Mst.
Aldehydes release zinc from proteins. A pathway from oxidative stress/lipid peroxidation to cellular functions of zinc
Authors: Hao Qiang , Maret Wolfgang
Published: August 2006

Oxidative stress, lipid peroxidation, hyperglycemia‐induced glycations and environmental exposures increase the cellular concentrations of aldehydes. A novel aspect of the molecular actions of aldehydes, e.g. acetaldehyde and acrolein, is their reaction with the cysteine ligands of zinc sites in proteins and concomitant zinc release. Stoichiometric amounts of acrolein release zinc from zinc–thiolate coordination sites in proteins such as metallothionein and alcohol dehydrogenase. Aldehydes also release zinc intracellularly in cultured human hepatoma (HepG2) cells and interfere with zinc‐dependent signaling processes such as gene expression and phosphorylation. Thus both acetaldehyde and acrolein induce the expression of metallothionein and modulate protein tyrosine phosphatase activity in a zinc‐dependent way. Since minute changes in the availability of cellular zinc have potent effects, zinc release is a mechanism of amplification that may account for many of the biological effects of aldehydes. The zinc‐releasing activity of aldehydes establishes relationships among cellular zinc, the functions of endogenous and xenobiotic aldehydes, and redox stress, with implications for pathobiochemical and toxicologic mechanisms.
Cell‐free translation systems for protein engineering
Authors: Shimizu Yoshihiro , Kuruma Yutetsu , Ying Bei‐Wen , Umekage So , Ueda Takuya
Published: August 2006

Cell‐free translation systems have developed significantly over the last two decades and improvements in yield have resulted in their use for protein production in the laboratory. These systems have protein engineering applications, such as the production of proteins containing unnatural amino acids and development of proteins exhibiting novel functions. Recently, it has been suggested that cell‐free translation systems might be used as the fundamental basis for cell‐like systems. We review recent progress in the field of cell‐free translation systems and describe their use as tools for protein production and engineering.
Cell‐free expression as an emerging technique for the large scale production of integral membrane protein
Authors: Klammt Christian , Schwarz Daniel , Löhr Frank , Schneider Birgit , Dötsch Volker , Bernhard Frank
Published: August 2006

Membrane proteins are highly underrepresented in structural data banks due to tremendous difficulties that occur upon approaching their structural analysis. Inefficient sample preparation from conventional cellular expression systems is in many cases the first major bottleneck. Preparative scale cell‐free expression has now become an emerging alternative tool for the high level production of integral membrane proteins. Many toxic effects attributed to the overproduction of recombinant proteins are eliminated by cell‐free expression as viable host cells are no longer required. A unique characteristic is the open nature of cell‐free systems that offers a variety of options to manipulate the reaction conditions in order to protect or to stabilize the synthesized recombinant proteins. Detergents or lipids can easily be supplemented and membrane proteins can therefore be synthesized directly into a defined hydrophobic environment of choice that permits solubility and allows the functional folding of the proteins. Alternatively, cell‐free produced precipitates of membrane proteins can efficiently be solubilized in mild detergents after expression. Highly valuable for structural approaches is the fast and efficient cell‐free production of uniformly or specifically labeled proteins. A considerable number of membrane proteins from diverse families like prokaryotic small multidrug transporters or eukaryotic G‐protein coupled receptors have been produced in cell‐free systems in high amounts and in functionally active forms. We will give an overview about the current state of the art of this new approach with special emphasis on technical aspects as well as on the functional and structural characterization of cell‐free produced membrane proteins.
A Metabonomics Study of the Hepatotoxicants Galactosamine, Methylene Dianiline and Clofibrate in Rats*
Authors: Ishihara Kenji , Katsutani Naruo , Aoki Toyohiko
Published: August 2006

Abstract: The efficacy of high‐resolution 1H nuclear magnetic resonance (1H‐NMR) spectroscopy‐based metabonomics was studied in a model of rat liver toxicity. Hepatotoxicities were induced in male rats using methylene dianiline, clofibrate and galactosamine. Twenty‐four‐hr urine from days 1 to 5 after treatment were subjected to 1H‐NMR evaluation of the biochemical effects. Blood were also taken at Days 2, 3 and 5 to examine biochemical changes associated with hepatotoxicities, and histopathological changes were evaluated at termination. Increases in liver enzymes were observed in animals treated with methylene dianiline or galactosamine, and histopathological analysis revealed changes associated with hepatobiliary damage and hepatocellular necrosis in methylene dianiline‐ and galactosamine‐treated animals, respectively. Principal component analysis and statistical Spotfire analyses were used to visualize similarities and differences in urine biochemical profiles produced by 1H‐NMR spectra. The biochemical effects of methylene dianiline and galactosamine were characterized by elevated levels of glucose, fructose, β‐hydroxybutyrate, alanine, acetoacetate, lactate and creatine and decreased levels of hippurate, 2‐oxoglutarate, citrate, succinate, trimethylamine‐N‐oxide, taurine and N‐acetylglutamate in rat urine. Clofibrate treatment elevated the levels of N‐methylnicotinamide and 3,4‐dihydroxymandelate and decreased the levels of 2‐oxoglutarate and N‐acetylaspartate. This work shows that combinations of 1H‐NMR and pattern recognition are powerful tools in the evaluation of the biochemical effects of xenobiotics in liver.
The Challenge of Predicting Drug Toxicityin silico
Authors: Vedani Angelo , Dobler Max , Lill Markus A.
Published: August 2006

Abstract:Poor pharmacokinetics, side effects and compound toxicity are frequent causes of late‐stage failures in drug development. A safein silicoidentification of adverse effects triggered by drugs and chemicals would be highly desirable as it not only bears economical potential but also spawns a variety of ecological benefits: sustainable resource management, reduction of animal models and possibly less risky clinical trials. In computer‐aided drug discovery, both existing and hypothetical compounds may be studied; the methods are fast, reproducible, and typically based on human bioregulators, making the question of transferability obsolete. In the recent past, our laboratory contributed towards the development ofin silicoconcepts (→ multi‐dimensional QSAR) and validated a series of “virtual test kits” based on the oestrogen, androgen, thyroid, and aryl hydrocarbon receptor (endocrine disruption, receptor‐mediated toxicity) as well as on the enzyme cytochrome P450 3A4 (metabolic transformations, drug‐drug interactions). The test kits are based on the three‐dimensional structure of their target protein (i.e.ERαβ,AR,TRαβ,CYP450) or a surrogate thereof (AhR) and were trained using a representative selection of 362 substances. Subsequent evaluation of 107 compounds different therefrom showed that binding affinities are predicted close to experimental uncertainty. These results suggest that our approach is suited for thein silicoidentification of adverse effects triggered by drugs and chemicals and encouraged us to compile an Internet Database for the virtual screening of drugs and chemicals for toxic effects.
Inactivation of α1‐Antiproteinase Induced by Phenylbutazone: Participation of Peroxyl Radicals and Hydroperoxide
Authors: Muraoka Sanae , Miura Toshiaki
Published: August 2006

Abstract: To clarify the action of a side‐effect of phenylbutazone, we investigated the inactivation of α1‐antiproteinase induced by phenylbutazone in the presence of horseradish peroxidase (HRP) and H2O2 (HRP‐H2O2). The activity of α1‐antiproteinase was rapidly lost during the interaction of phenylbutazone with HRP‐H2O2 under aerobic conditions. Phenylbutazone showed a marked spectral change under aerobic conditions but not under anaerobic conditions. Spin trap agents were very effective in inhibiting α1‐antiproteinase inactivation induced by phenylbutazone. Oxidation of phenylbutazone was stopped by catalase, but the inactivation reaction of α1‐antiproteinase proceeded even after removal of H2O2 in the reaction mixture. Formation of the peroxidative product from phenylbutazone was detected by iodometric assay. These results indicate that both peroxyl radicals and the peroxidative product of phenylbutazone participated in the inactivation of α1‐antiproteinase. Other anti‐inflammatory drugs did not inactivate α1‐antiproteinase during interaction with HRP‐H2O2. Inactivation of α1‐antiproteinase may contribute to serious side effects of phenylbutazone.
Protective Effect of Ebselen, a Selenoorganic Drug, against Gentamicin‐Induced Renal Damage in Rats
Authors: Dhanarajan R. , Abraham Premila , Isaac Bina
Published: August 2006

Abstract: Gentamicin is an antibiotic that is widely used against serious and life‐threatening gram‐negative infections. However, its clinical use is limited by its nephrotoxicity. Oxidative stress and nitrosative stress are reported to play important role in gentamicin nephrotoxicity. In the present study we investigated whether ebselen, an inhibitor of oxidative stress and nitrosative stress prevents or reduces gentamicin‐induced renal damage in the rat. For this purpose male Wistar rats were divided into five groups and treated as follows. Group 1 (control group): dimethyl sulphoxide, intraperitoneally, Group 2: Gentamicin 100 mg/kg b.wt. subcutaneously, Group 3: 5 mg/g b.wt. ebselen intraperitoneally, Group 4: 2.5 mg/kg b.wt. ebselen followed by 100 mg/kg b.wt. gentamicin subcutaneously one hour later, and Group 5: 5 mg/kg b.wt. of ebselen followed by 100 mg/kg b.wt. gentamicin one hour later for four consecutive days. Nephrotoxicity was evaluated histopathologically by light microscopy, and biochemically by the measurement of the plasma creatinine and urea levels. Parameters of oxidative stress such as reduced glutathione, malondialdehyde, and activities of superoxide dismutase and glutathione peroxidase were measured in the kidney. Serum nitrite and nitrate were measured as indicators of nitrosative stress. Treatment of rats with gentamicin resulted in statistically significant reduction in reduced glutathione levels (51%) and the activities of antioxidant enzymes superoxide dismutase (56%) and glutathione peroxidase (39%) as compared with the controls in the kidneys. Renal malondialdehyde level was increased significantly (43%) as compared with the controls. Plasma creatinine levels, urea levels and nitrite levels were significantly increased (4, 4.5 and 160% times respectively) as compared with the controls. Histologically, damage to the renal cortex and medulla was observed moderate to severe tubular necrosis and glomerular congestion. Pretreatment with 2.5 mg/kg b.wt. ebselen prevented gentamicin induced damage to medulla; however, renal cortex showed mild damage and biochemically indicators of oxidative stress and nitrosative stress were significantly reduced. Pretreatment with 5 mg/kg b.wt. ebselen prevented gentamicin‐induced oxidative damage and nitrosative damage and renal damage almost completely in 78% of the rats, in the other 22% of the rats, ebselen pretreatment reduced gentamicin‐induced renal damage. The results of the present study suggest that ebselen may be useful as a nephroprotective agent.
Proton Pump Inhibitors and Helicobacter pylori Gastritis: Friends or Foes?
Authors: Kuipers Ernst J.
Published: August 2006

Abstract: H. pylori gastritis and gastric acid closely interact. In H. pylori‐positive patients, profound acid suppressive therapy induces a corpus‐predominant pangastritis, which is associated with accelerated corpus gland loss and development of atrophic gastritis. Both corpus‐predominant and atrophic gastritis have been associated with an increased risk of development of gastric cancer. H. pylori eradication leads to resolution of gastritis and may induce partial regression of pre‐existent gland loss. H. pylori eradication does not aggravate GERD nor does it impair the efficacy of proton pump inhibitor maintenance therapy for this condition. This is the background of the advise within the European guidelines for the management of H. pylori infection to offer an H. pylori test and treat policy to patients who require proton pump inhibitor maintenance therapy for GERD. As such a policy fully reverses H. pylori pangastritis even in patients who have been treated for years with proton pump inhibitors, there is no need to eradicate H. pylori before the start of proton pump inhibitors. In fact, the somewhat slower initial response of H. pylori‐negative GERD patients to proton pump inhibitor therapy and the fact that many GERD patients will only require short‐term therapy suggests to first start the proton pump inhibitor, and only test and treat when maintenance therapy needs to be prescribed. Such considerations prevent the persistent presence of active corpus‐predominant gastritis in proton pump inhibitor‐treated reflux patients without impairing the clinical efficacy of treatment
The Protective Effect of Captopril on Nicotine‐Induced Endothelial Dysfunction in Rat
Authors: Luo Hong‐Li , Zang Wei‐Jin , Lu Jun , Yu Xiao‐Jiang , Lin Yuan‐Xi , Cao Yong‐Xiao
Published: August 2006

Abstract: This study was designed to examine the in vivo and in vitro effects of captopril, an angiotensin‐converting enzyme inhibitor, on nicotine‐induced endothelial dysfunction in rats. Endothelial dysfunction was induced by exposing isolated rat mesenteric arteries to nicotine (0.01, 0.1, or 1 mM) for 24 hr using an organ culture system, or by treating rats with nicotine (2 mg/kg/day, intraperitoneally) for 4 weeks. The protective effects of captopril were tested by exposing isolated mesenteric arteries to captopril (0.01, 0.03, or 0.1 mM) + nicotine (0.1 mM) for 24 hr, or by treating rats with captopril (3 mg/kg/day, intravenously) + nicotine (2 mg/kg/day, intraperitoneally) for 4 weeks. Exposure of the isolated mesenteric arteries to nicotine induced a significant concentration ‐dependent inhibition of endothelium‐dependent relaxation. Co‐culture of segments of mesenteric artery with captopril (0.03 or 0.1 mM) attenuated the nicotine‐induced impairment of vasorelaxation in a dose‐dependent manner. Administration of nicotine to rats for 4 weeks significantly impaired endothelium‐dependent relaxation compared with control rats. This impairment was accompanied by a reduction in nitrite/nitrate, nitric oxide (NO) synthase (NOS), and superoxide dismutase (SOD) activities in the serum and aorta. Chronic captopril treatment not only improved the impairment of endothelium‐dependent relaxation, but also prevented the reduction of nitrite/nitrate contents and of NOS and SOD activities in the serum and aorta. However, there were no significant differences in serum angiotensin‐converting enzyme activity among the three groups. These results indicate that captopril can be used to attenuate nicotine‐induced endothelial dysfunction, an effect that may be related not only to antioxidation, but also to enhancing NO production by preventing the decrease in NOS.
Grape Seed Extract Prevents Gentamicin‐Induced Nephrotoxicity and Genotoxicity in Bone Marrow Cells of Mice
Authors: El‐Ashmawy Ibrahim M. , El‐Nahas Abeer F. , Salama Osama M.
Published: August 2006

Abstract: The protection conferred by grape seed extract against gentamicin‐induced nephrotoxicity and bone marrow chromosomal aberrations have been evaluated in adult Swiss albino mice. The activity of reduced glutathione peroxidase (GSH peroxidase), the levels of glutathione (GSH) and lipid peroxidation as malondialdehyde (MDA) in the kidneys homogenates, serum urea and creatinine were measured, and in addition the changes in kidney histology and bone marrow chromosomes were investigated. Gentamicin (80 mg/kg b.wt. intraperitoneally for 2 weeks) induced kidney damage as indicated from a pronounced changes in kidney histology, a significant increase in serum urea and creatinine and MDA content in the kidney homogenate. While the activity of the antioxidant enzyme GSH peroxidase and the level of GSH were significantly decreased. Gentamicin induced genotoxicity indicated by increased the number of aberrant cells and different types of structural chromosomal aberrations (fragment, deletion and ring chromosome) and showed no effect on mitotic activity of the cell. Pretreatment with grape seed extract (7 days) and simultaneously (14 days) with gentamicin significantly protected the kidney tissue by ameliorating its antioxidant activity. Moreover, grape seed extract significantly protected bone marrow chromosomes from gentamicin induced genotoxicity by reducing the total number of aberrant cells, and different types of structural chromosomal aberrations. It could be concluded that grape seed extract acts as a potent antioxidant prevented kidney damage and genotoxicity of bone marrow cells.
Gene expression patterns in isolated keloid fibroblasts
Authors: Satish Latha , Lyons‐Weiler James , Hebda Patricia A. , Wells Alan
Published: August 2006

Keloid scars after skin trauma are a significant clinical problem, especially in black populations, in which the incidence of keloids has been estimated at 4–16%. Keloids are abnormal dermal proliferative scars secondary to dysregulated wound healing. Despite several biochemical studies on the role of extracellular matrix proteins and growth factors during keloid formation, we still do not know what molecules and signals induce this change. Fibroblasts are thought to be the major inductive cell for keloid scar formation. The aim of this study was to identify gene expression patterns that characterize keloid fibroblasts; identifying such genetic disequilibrium may shed light on the molecular signaling events responsible for keloid formation. In this study, we performed gene expression analysis of fibroblasts isolated from keloid lesions from three individuals in comparison with the fibroblasts isolated from normal skin using the Affymetrix U133a chip (22,284 genes and expression sequence tags). We found through J5 test score expression analysis that among 22,284 genes, there were 43 genes that were overexpressed and five genes were underexpressed in keloid fibroblasts when compared with dermal fibroblasts from persons without keloids. The overexpression of three genes not previously reported as being up‐regulated in keloids (annexin A2, Transgelin, and RPS18) was confirmed by real‐time polymerase chain reaction. Certain overexpressed genes were similar to previous biochemical observations on the protein levels of these overexpressed genes during keloid formation. We also report for the first time that a few tumor‐related genes are overexpressed in keloid fibroblasts.
Efficacy and cost‐effectiveness of a high‐powered parallel waterjet for wound debridement
Authors: Granick Mark S. , Posnett John , Jacoby, BS Michael , Noruthun Shyam , Ganchi Parham A. , Datiashvili Ramazi O.
Published: August 2006

Current concepts of wound healing acknowledge the essential role of wound bed preparation in achieving a wound with good healing potential. Critical to wound bed preparation is the removal of necrosis, unhealthy tissue, foreign matter, and infection. One of the accepted methods of wound bed preparation is surgery. The high‐power parallel waterjet is a new surgical device, which allows the operator to remove very precisely undesirable tissue and debris with maximal preservation of viable tissue. A retrospective study was performed to evaluate the efficacy, safety, and economic impact of using this technique of surgical debridement. Forty patients who had waterjet debridements were compared with 22 patients with matched wounds who had conventional surgical debridement. The waterjet group had significantly fewer procedures (p<0.002) than the conventional group. Based on these outcomes, the use of the new device in appropriate patients is expected to lead to cost savings of approximately $1,900 per patient.
Identification of a novel FcγRIII receptor that is up‐regulated in fetal wound healing
Authors: Teusner Jacqueline T. , Goddard Chris , Belford David A. , Dunaiski Vera , Powell Barry C.
Published: August 2006

The mid‐gestation fetus is able to heal skin wounds rapidly and without scarring, an ability that is lost as development proceeds. The aim of this study was to identify novel genes involved in this process. We established an ex vivo wound model from embryonic rats and showed that over 72 hours, embryonic day 17 wounds reepithelialized and closed whereas day 19 wounds did not. To investigate the molecular basis of this phenomenon we analyzed changes in gene expression using differential display polymerase chain reaction. We characterized one transcript that was strongly up‐regulated in the healing response of wounded, day 17 skin. It encodes a protein of 249 amino acids with striking similarity to the human low‐affinity receptor for the Fc portion of IgG (FcγRIII), suggesting that it is a novel member of the FcγR family, which we named FcγRIII‐X. A wound‐healing timecourse shows that FcγRIII‐X was up‐regulated in healing, wounded day 17 skin but not in nonhealing, wounded day 19 skin and that its up‐regulation was accelerated in skin with multiple wounds. We suggest that up‐regulation of FcγRIII‐X may contribute to scarless healing of fetal skin.