bluuebunny/crossref_metadata_2025 huggingface dataset:
Lack of In Vivo Correlation Between Indinavir and Saquinavir Exposure and Cytochrome P450 3A Phenotype as Assessed with Oral Midazolam as a Phenotype Probe
Authors: Robertson Sarah M. , Formentini Elizabeth , Alfaro Raul M. , Falloon M.S. Judith , Penzak Scott R.
Published: July 2006

Study Objective. To investigate a potential correlation between exposure to oral midazolam, a commonly used cytochrome P450 (CYP) 3A probe, and saquinavir and indinavir exposure.

Design. Open‐label, prospective, pharmacokinetic study.

Setting. Outpatient research center.

Subjects. Thirty‐six healthy volunteers aged 22–50 years.

Intervention. Subjects received a single oral dose of midazolam 8 mg; 4 hours later, blood was drawn to determine their serum midazolam concentrations. Midazolam phenotyping was followed by successive administration of the protease inhibitors indinavir and saquinavir, with blood sampling and pharmacokinetic analyses performed at steady state.

Measurements and Main Results. Pharmacokinetic parameters of each protease inhibitor were evaluated to assess for a potential relationship with 4‐hour concentrations of midazolam. No correlations between phenotype results for midazolam and any pharmacokinetic parameter for indinavir or saquinavir were identified (r2=0.00002–0.073). When the results were analyzed based on race, significant correlations were identified in five African‐American subjects, including correlations between 4‐hour midazolam levels and apparent oral clearance of saquinavir (r2=0.734, p=0.064), area under the plasma concentration–time curve from 0–8 hours (r2=0.914, p=0.011), minimum concentration (r2=0.857, p=0.024), and maximum concentration (r2=0.969, p=0.002). These findings for African‐American subjects were not seen with indinavir. No correlation was found between indinavir and saquinavir pharmacokinetic parameters (r2=0.017–0.261).

Conclusion. Oral midazolam was not a useful probe for predicting saquinavir or indinavir exposure at steady state. Reasons for the lack of correlation likely included differences between midazolam and protease inhibitor P‐glycoprotein specificity, differences in the relative contribution of CYP3A5‐mediated metabolism, and/or variation in intestinal and hepatic CYP3A specificity. The strong correlation between midazolam phenotype and pharmacokinetic parameters for saquinavir in African‐American subjects indicated a racial difference in one or more of these confounding variables.
Drug Copayment and Adherence in Chronic Heart Failure: Effect on Cost and Outcomes
Authors: Cole J. Alexander , Norman Heather , Weatherby Lisa B. , Walker Alexander M.
Published: July 2006

Study Objective. To measure the association among prescription copayment, drug adherence, and subsequent health outcomes among patients with chronic heart failure (CHF).

Design. Retrospective cohort study.

Data Source. Database of a large, national health insurance plan.

Patients. Patients with CHF receiving commercial and Medicare supplemental benefits.

Measurements and Main Results. We estimated adherence to therapy with β‐blockers or angiotensin‐converting enzyme (ACE) inhibitors in 2002 by using the medication possession ratio, an estimate of the proportion of days a patient was exposed to a drug while taking a drug regimen. For 2003, we measured the annualized total cost of health care and identified hospitalizations with a diagnostic code for CHF. We used a two‐stage regression approach to model the association among copayment, adherence, and patient outcomes. For patients taking ACE inhibitors, a $10 increase in copayment was associated with a 2.6% decrease in the medication possession ratio (95% confidence interval [CI] 2.0–3.1%). This change in adherence was associated with a predicted 0.8% decrease in medical costs (95% CI‐4.2–2.5%) but a predicted 6.1% increase in the risk of hospitalization for CHF (95% CI 0.5–12.0%). Among patients taking β‐blockers, a $10 increase in copayment was associated with a 1.8% decrease in the medication possession ratio (95% CI 1.4–2.2%). This change in adherence was associated with a predicted 2.8% decrease in medical costs (95% CI −5.9–0.1%) and a predicted 8.7% increase in the risk of hospitalization for CHF (95% CI 3.8–13.8%).

Conclusion. Among patients with CHF, higher drug copayments were associated with poorer adherence. The change was relatively small and did not affect predicted total health care costs, but it was sufficient to increase the predicted risk of hospitalization for CHF.
Golf‐Inhibiting Gynecomastia Associated with Atorvastatin Therapy
Authors: Hammons Kimberly B. , Edwards Rebecca F. , Rice William Y.
Published: July 2006

Gynecomastia can have a significant emotional and social impact on men. Although numerous drug therapies may cause this condition, we found no documented cases of gynecomastia associated with atorvastatin. We describe the development of breast enlargement and tenderness in a 52‐year‐old Caucasian man 6 months after his simvastatin therapy had been switched to atorvastatin. His symptoms ultimately interfered with his ability to play golf and participate in other activities. These problems resolved after atorvastatin discontinuation and did not recur despite resumption of simvastatin therapy. The gynecomastia in this patient represented a possible adverse effect of atorvastatin according to the Naranjo adverse drug reaction probability scale. The mechanism may be atorvastatin's theoretical suppression of adrenal or gonadal steroid production through effects on cholesterol synthesis. Based on this and other case reports, 3‐hydroxy‐3‐methylglutaryl coenzyme A reductase inhibitors (statins) should be considered as a potential cause when evaluating otherwise unexplainable cases of gynecomastia in patients taking these drugs.
Bell's Palsy Associated with Linezolid Therapy: Case Report and Review of Neuropathic Adverse Events
Authors: Thai Xia C. , Bruno‐Murtha Lou Ann
Published: July 2006

Bell's palsy is one of the most common neurologic disorders affecting the seventh cranial nerve. Several disease states have been associated with facial paralysis. Drugs, however, have been rarely implicated as an etiology. We describe a 49‐year‐old man who developed peripheral facial paralysis after 3 weeks of linezolid therapy, along with recurrence of symptoms on rechallenge. He had insulin‐dependent diabetes mellitus and a longstanding history of bilateral diabetes‐related foot problems. After hospitalization, debridement, and vancomycin therapy for methicillin‐resistant Staphylococcus aureus osteomyelitis, the patient was discharged to home with oral linezolid therapy. On day 23 of linezolid therapy, he developed signs and symptoms that were consistent with Bell's palsy. Linezolid was discontinued; the Bell's palsy gradually improved, with complete resolution occurring at month 3. On rechallenge with linezolid for recurrent osteomyelitis, the patient developed a second episode of Bell's palsy within a similar time frame as in the first episode. Assessment of causality using the Naranjo adverse drug reaction probability scale revealed a probable relationship between this adverse drug event and linezolid therapy. Clinicians should be aware that Bell's palsy may be another neuropathic adverse effect associated with linezolid.
Monomolecular‐Layer Ba5Ta4O15 Nanosheets: Synthesis and Investigation of Photocatalytic Properties
Authors: Xu T.‐G. , Zhang C. , Shao X. , Wu K. , Zhu Y.‐F.
Published: July 2006

Monomolecular‐layer perovskite Ba5Ta4O15 nanosheets with hexagonal structure have been synthesized by a hydrothermal method. The thickness of the nanosheets is about 1.1 nm, which corresponds to a monolayer of Ba5Ta4O15 molecules, with the lateral size ranging from 50 to 200 nm. The optimal conditions for the formation of the nanosheets are maintaining the reactants above 270 °C for 24 h. A dissolution–recrystallization mechanism is suggested based on observations of the factors that influence nanosheet formation, such as reaction time, temperature, and basicity. Formation of Ba5Ta4O15 nanosheets takes precedence over other nanostructures under high concentrations of OH– because the hindering effect of OH– ions on the c‐axis growth is strong. Thus, the extended growth rate of polyhedrons on one monolayer is much faster than the superposition rate of the monolayer, and the crystal grows more easily along the a‐ and b‐planes. The Ba5Ta4O15 nanosheets show a high photocatalytic activity in the degradation of Rhodamine B and gaseous formaldehyde. The layered perovskite probably affects the photocatalytic activity by promoting the charge separation and delocalization of photogenerated electrons and holes.
Spectroscopic pH measurement for high temperatures, pressures and ionic strength
Authors: Raghuraman B. , Gustavson G. , Mullins O. C. , Rabbito P.
Published: July 2006

The optical spectroscopic technique using pH sensitive dyes has been extended to measure pH in high‐temperature, pressure, and ionic strength environments. Lack of standard calibrating buffers for these conditions necessitated a two‐step process for development of this technique. The first step involved chemical equilibrium modeling to predict pH values of simple buffer salt systems at high temperatures, pressures and ionic strengths. The second step involved using these modeled buffer solutions as calibration solutions for the dye chemistry and equilibrium dissociation constant. Optical spectra of dye in these buffer solutions were used to calibrate the dye equilibrium dissociation constant. Results are presented for 293°K to 373°K temperature range, 0.101325–65.5 MPa pressure range and 0–3 mol/kg water ionic strength range. Once the dye equilibrium dissociation constant has been characterized, pH of unknown solutions can be obtained from their dye spectra. This method has been successfully employed for high‐temperature and high‐pressure measurements of oilfield waters. © 2006 American Institute of Chemical Engineers AIChE J, 2006
Postpartum acquired factor VIII inhibitors
Authors: Franchini Massimo
Published: July 2006

The development of factor VIII autoantibodies is a rare but severe complication of pregnancy. Although the natural history of postpartum acquired hemophilia A is usually benign, with a high percentage of spontaneous remissions and a low mortality, its quick recognition is important to control bleeding episodes.

Based on an analysis of the literature, this review presents the current knowledge on the pathogenesis, diagnosis, epidemiology, natural history, clinical manifestations, and therapeutic management of postpartum acquired hemophilia A. Am. J. Hematol., 2006. © 2006 Wiley‐Liss, Inc.
Flight‐oogenesis syndrome in a blood‐sucking bug: Biochemical aspects of lipid metabolism
Authors: Oliveira Giselle A. , Baptista Daniela L. , Guimarães‐Motta Horacio , Almeida Igor C. , Masuda Hatisaburo , Atella Georgia C.
Published: July 2006

Lipophorin (Lp), either labeled in diacylglycerol moiety with [3H]‐Palmitic acid or in phospholipid moiety with 32Pi, was injected into Rhodnius prolixus females. Insects were induced to flight for different times. In just a few minutes of flight, the transfer of radioactivity to ovaries decreased, accompanied by its increase to flight muscles. After one hour of flight, Lp density was higher (1.132 g/mL) than before flight (1.116 g/mL). Lp purified from insects after flight was analyzed by gel filtration chromatography and a polyacrylamide gel pore limit electrophoresis. Both analyses demonstrated a decrease in Lp molecular mass after flight but no changes in apoLp‐III amounts were observed. Time‐course experiments showed that only 30 min of flight are required for the detection of changes in Lp density and molecular mass. About the same time of rest is necessary for Lp density and molecular mass to return to the baseline value. The lipid content from Lp particles, determined by high‐performance thin‐layer chromatography (HPTLC), showed a decrease in total lipids after flight. At the same time, an increase of many classes of lipids was observed in flight muscles except for triacylglycerol, which was reduced. The increase of flight muscle lipids was accompanied by a decrease of the ovaries lipid content. The insects subjected to daily exhaustive flight showed a significant decrease in total number of eggs produced. But insects subjected to a single exhaustive flight showed only a small reduction in total number of eggs. Lp density variation during the flight activity of Rhodnius prolixus females is discussed in association with physiological events such as oogenesis. Arch Insect Biochem Physiol 62:164–175, 2006. © 2006 Wiley‐Liss, Inc.
Identification in human osteoarthritic chondrocytes of proteins binding to the novel regulatory site AGRE in the human matrix metalloprotease 13 proximal promoter
Authors: Fan Zhiyong , Tardif Ginette , Boileau Christelle , Bidwell Joseph P. , Geng Changshan , Hum David , Watson Alexander , Pelletier Jean‐Pierre , Lavigne Martin , Martel‐Pelletier Johanne
Published: July 2006

Objective

Matrix metalloprotease 13 (MMP‐13) plays a major role in osteoarthritic (OA) processes. We previously identified the AG‐rich element (AGRE) regulatory site (GAAAAGAAAAAG) in the proximal promoter of this gene. Electrophoretic mobility shift assays (EMSAs) done with nuclear extracts from OA chondrocytes showed the presence of 2 AGRE protein–binding complexes, the formation of which depended on the pathophysiologic state (high or low) of the cells; the low OA (L‐OA) chondrocytes have low MMP‐13 basal levels and high interleukin‐1β (IL‐1β) inducibility, and the high OA (H‐OA) chondrocytes have high MMP‐13 basal levels and low IL‐1β inducibility. In this study, we sought to determine the importance of individual AGRE bases in promoter activity and to identify AGRE binding proteins from L‐OA and H‐OA chondrocyte complexes.

Methods

Promoter activity was determined following transient transfection into human OA chondrocytes. AGRE binding proteins were identified by mass spectroscopy.

Results

Individual mutations of the AGRE site differentially modulated promoter activity, indicating that the intact AGRE site is required for optimal MMP‐13 expression. Damage‐specific DNA binding protein 1 (DDB‐1) was identified in the L‐OA chondrocyte–binding complex. EMSA experiments performed with the mutation of the left AGRE site (GTGCTGAAAAAG) and nuclear extracts of L‐OA chondrocytes reproduced the pattern seen in the H‐OA chondrocytes. Mass spectroscopy identified p130cas as one of the proteins in this complex. Supershift experiments showed the presence of p130cas and nuclear matrix transcription factor 4 (NMP‐4) in the wild‐type AGRE/H‐OA chondrocyte complex.

Conclusion

These data suggest that the binding of p130cas and NMP‐4 to the AGRE site regulates MMP‐13 expression and may trigger the change in human chondrocytes from the L‐OA state to the H‐OA state.
RANTES and monocyte chemoattractant protein 1 as sensitive markers of disease activity in patients with juvenile rheumatoid arthritis: A six‐year longitudinal study
Authors: Yao Tsung‐Chieh , Kuo Ming‐Ling , See Lai‐Chu , Ou Liang‐Shiou , Lee Wen‐I , Chan Chin‐Kan , Huang Jing‐Long
Published: July 2006

Objective

To longitudinally investigate serum and synovial fluid (SF) levels of RANTES and monocyte chemoattractant protein 1 (MCP‐1) as well as in vitro migration of mononuclear cells toward SF in patients with juvenile rheumatoid arthritis (JRA).

Methods

Serum and SF levels of RANTES and MCP‐1 were determined by enzyme‐linked immunosorbent assay. Chemotaxis was performed using the modified Boyden chamber method.

Results

Serum RANTES levels were significantly increased in all onset types of JRA, with the highest levels present in systemic‐onset JRA. Serum MCP‐1 levels were significantly elevated in patients with systemic‐onset JRA and were associated with current systemic features. Although serum levels of RANTES and MCP‐1 decreased significantly after treatment, RANTES and MCP‐1 levels during disease remission were still significantly higher in JRA patients than in controls. A relationship was found between serum RANTES levels during remission and the duration of clinical remission, with low levels being associated with prolonged clinical remission and high levels with shorter clinical remission. Serum RANTES levels correlated with C‐reactive protein concentrations, hemoglobin values, white blood cell (WBC) counts, and platelet counts, whereas serum MCP‐1 levels correlated with WBC counts and serum ferritin levels. Levels of RANTES and MCP‐1 in SF were elevated as compared with levels in serum. SF chemotactic activity for mononuclear leukocytes was significantly inhibited by either anti‐RANTES or anti–MCP‐1 antibody.

Conclusion

RANTES is a key molecule in the pathogenesis of all onset groups of JRA, whereas MCP‐1 is particularly important in systemic‐onset JRA. Serum levels of these CC chemokines represent more highly sensitive markers of disease activity than conventional markers of inflammation.
Relationship between serum NR2a antibodies and cognitive dysfunction in systemic lupus erythematosus
Authors: Harrison Melanie J. , Ravdin Lisa D. , Lockshin Michael D.
Published: July 2006

Objective

To assess the association between serum NR2a antibodies and cognitive dysfunction in systemic lupus erythematosus (SLE).

Methods

The study population consisted of English‐speaking adults who met American College of Rheumatology (ACR) criteria for SLE and had at least 1 serum sample stored in the Hospital for Special Surgery Autoimmune Registry and Repository. Demographic and clinical information was obtained, and patients completed the neuropsychological test battery recommended by the ACR, the Center for Epidemiologic Studies Depression Scale, and the Spielberger State‐Trait Anxiety Inventory. Cognitive impairment was defined as scores >1.5 SD below the mean of age‐matched published normative data on at least 2 neuropsychological tests. Sera were tested for NR2a antibodies by enzyme‐linked immunosorbent assay. Performance on neuropsychological tests was compared between NR2a‐positive and NR2a‐negative patients.

Results

Of the 93 patients, 24 (25.8%) were positive for NR2a antibodies. Of the 48 patients who were cognitively impaired based on test results, 31% were positive for NR2a antibodies, compared with 20% of those who were not cognitively impaired (P = 0.24). Among antibody‐positive patients, the mean ± SD number of neuropsychological tests with abnormal results was 2.3 ± 2.2, compared with 2.0 ± 1.8 in the antibody‐negative group (P = 0.59). Similar nonsignificant differences were found when impairment was defined using a more stringent definition (i.e., test scores >2.0 SD below the mean) and using a neuropsychologist's clinical ratings. No association was detected between NR2a antibody positivity and depressive symptoms (P = 0.73) or anxiety (P = 0.42).

Conclusion

No significant association was found between NR2a antibody positivity and cognitive dysfunction, depressive symptoms, or anxiety. These results indicate that the presence of these antibodies alone does not have a direct effect on cognitive functioning or any other neuropsychiatric manifestation of SLE.
Inhibition of angiogenesis by interleukin‐4 gene therapy in rat adjuvant‐induced arthritis
Authors: Haas Christian S. , Amin M. Asif , Allen Brittany B. , Ruth Jeffrey H. , Haines G. Kenneth , Woods James M. , Koch Alisa E.
Published: July 2006

Objective

Interleukin‐4 (IL‐4) can modulate neovascularization. In this study, we used a gene therapy approach to investigate the role of IL‐4 in angiogenesis in rat adjuvant‐induced arthritis (AIA), a model for rheumatoid arthritis.

Methods

Rats received an adenovirus producing IL‐4 (AxCAIL‐4), a control virus without insert, or control vehicle (phosphate buffered saline) intraarticularly before arthritis onset. At peak onset of arthritis, rats were killed. Vascularization was determined in the synovial tissue, and correlations with inflammation were assessed. Ankle homogenates were used in angiogenesis assays in vitro and in vivo, and protein levels of cytokines and growth factors were assessed by enzyme‐linked immunosorbent assay. Synovial tissue expression of αv integrins was determined by immunohistochemistry.

Results

IL‐4 induced a reduction in synovial tissue vessel density, which was paralleled by a decrease in inflammation. AxCAIL‐4 joint homogenates significantly (P< 0.05) inhibited both endothelial cell (EC) migration and tube formation in vitro. Similarly, AxCAIL‐4 inhibited capillary sprouting in the rat aortic ring assay, and vessel growth in the in vivo Matrigel plug assay. The angiostatic effect occurred despite high levels of vascular endothelial growth factor (VEGF), and was associated with down‐regulation of the proangiogenic cytokines IL‐18, CXCL16, and CXCL5 and up‐regulation of the angiogenesis inhibitor endostatin. Of interest, AxCAIL‐4 also resulted in decreased EC expression of the αv and β3 integrin chains.

Conclusion

In rat AIA, IL‐4 reduces synovial tissue vascularization via angiostatic effects, mediates inhibition of angiogenesis via an association with altered pro‐ and antiangiogenic cytokines, and may inhibit VEGF‐mediated angiogenesis and exert its angiostatic role in part via αvβ3 integrin. This knowledge of the specific angiostatic effects of IL‐4 may help optimize target‐oriented treatment of inflammatory arthritis.
Sample Size Recalculation in Internal Pilot Study Designs: A Review
Authors: Friede Tim , Kieser Meinhard
Published: July 2006

The adequacy of sample size is important to clinical trials. In the planning phase of a trial, however, the investigators are often quite uncertain about the sizes of parameters which are needed for sample size calculations. A solution to this problem is mid‐course recalculation of the sample size during the ongoing trial. In internal pilot study designs, nuisance parameters are estimated on the basis of interim data and the sample size is adjusted accordingly. This review attempts to give an overview on the available methods. It is written not only for biometricians who are already familar with the the topic and wish to update their knowledge but also for users new to the subject. (© 2006 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Construction of multi‐layered cardiomyocyte sheets using magnetite nanoparticles and magnetic force
Authors: Shimizu Kazunori , Ito Akira , Lee Jong‐Kook , Yoshida Tatsuro , Miwa Keiko , Ishiguro Hisaaki , Numaguchi Yasushi , Murohara Toyoaki , Kodama Itsuo , Honda Hiroyuki
Published: July 2006

Heart tissue engineering requires construction of three‐dimensional (3‐D) tissues composed of cardiomyocytes (CMs) that are tightly connected to each other. The aim of this study was to construct “scaffold‐less” multi‐layered 3‐D CM sheets using magnetic force‐based tissue engineering (Mag‐TE) and to evaluate the cell‐to‐cell functional connections within the CM sheets. Original magnetite cationic liposomes (MCLs) with a positive surface charge (which facilitate adsorption to the target cell surface) were taken up by CMs that were isolated from 2‐day‐old Wistar rats. When MCLs were added to the medium of CMs at magnetite concentrations of 25, 50, and 100 pg per cell, subsequent measurements showed that 7.2, 13.2, and 27.3 pg of magnetite were taken up per cell, respectively, after 4 h incubation at 37°C. Further, no toxicity was observed after a 24 h incubation period. Using magnetically labeled CMs (magnetite concentration, 100 pg/cell), multi‐layered CM sheets were constructed. Immunofluorescent staining of connexin43 demonstrated the presence of gap junctions within the CM sheets that were constructed by Mag‐TE. Moreover, electrical connections within the CM sheets constructed by Mag‐TE were confirmed using extracellular potential mapping. These results indicate that Mag‐TE is a viable methodology for heart tissue engineering. Biotechnol. Bioeng. 2007;96:803–809. © 2006 Wiley Periodicals, Inc.
A kinetic model describing Shewanella oneidensis MR‐1 growth, substrate consumption, and product secretion
Authors: Tang Yinjie J. , Meadows Adam L. , Keasling Jay D.
Published: July 2006

Aerobic growth of Shewanella oneidensis MR‐1 in minimal lactate medium was studied in batch cultivation. Acetate production was observed in the middle of the exponential growth phase and was enhanced when the dissolved oxygen (DO) concentration was low. Once the lactate was nearly exhausted, S. oneidensis MR‐1 used the acetate produced during growth on lactate with a similar biomass yield as lactate. A two‐substrate Monod model, with competitive and uncompetitive substrate inhibition, was devised to describe the dependence of biomass growth on lactate, acetate, and oxygen and the acetate growth inhibition across a broad range of concentrations. The parameters estimated for this model indicate interesting growth kinetics: lactate is converted to acetate stoichiometrically regardless of the DO concentration; cells grow well even at low DO levels, presumably due to a very low Km for oxygen; cells metabolize acetate (maximum specific growth rate, µmax,A of 0.28 h−1) as a single carbon source slower than they metabolize lactate (µmax,L of 0.47 h−1); and growth on acetate is self‐inhibiting at a concentration greater than 10 mM. After estimating model parameters to describe growth and metabolism under six different nutrient conditions, the model was able to successfully estimate growth, oxygen and lactate consumption, and acetate production and consumption under entirely different growth conditions. Biotechnol. Bioeng. 2007;96: 125–133. © 2006 Wiley Periodicals, Inc.
Inhibitory effects of nitrogen oxides on a mixed methanogenic culture
Authors: Tugtas A. Evren , Pavlostathis Spyros G.
Published: July 2006

The effect of nitrate, nitrite, nitric oxide (NO), and nitrous oxide on a mixed, mesophilic (35°C) methanogenic culture was investigated. Short‐term inhibition assays were conducted at a concentration range of 10–350 mg N/L nitrate, 17–500 mg N/L nitrite, 0.02–0.8 mg N/L aqueous NO, and 19–191 mg N/L aqueous nitrous oxide. Simultaneous methane production and N‐oxide reduction was observed in 10 and 30 mg N/L nitrate and 0.02 mg N/L aqueous NO‐amended cultures. However, addition of N‐oxide resulted in immediate cessation of methanogenesis in all other cultures. Methanogenesis completely recovered subsequent to the complete reduction of N‐oxides to nitrogen gas in all N‐oxide‐amended cultures, with the exception of the 500 mg N/L nitrite‐ and 0.8 mg N/L aqueous NO‐amended cultures. Partial recovery of methanogenesis was observed in the 500 mg N/L nitrite‐amended culture in contrast to complete inhibition of methanogenesis in the 0.8 mg N/L aqueous NO‐amended culture. Accumulation of volatile fatty acids was observed in both cultures at the end of the incubation period. Among all N‐oxides, NO exerted the most and nitrate exerted the least inhibitory effect on the fermentative/methanogenic consortia. The effect of multiple additions of nitrate (300 mg N/L) on the same methanogenic culture was also investigated. Long‐term exposure of the methanogenic culture to nitrate resulted in an increase of N‐oxide reduction rates and decrease of methane production rates, which was attributed to changes in the microbial community structure due to nitrate addition. Biotechnol. Bioeng. 2007;96: 444–455. © 2006 Wiley Periodicals, Inc.
Activating an Enzyme by an Engineered Coiled Coil Switch
Authors: Yuzawa Satoshi , Mizuno Toshihisa , Tanaka Toshiki
Published: July 2006

We designed a de novo protein based on a circular permutant of RNaseT1, in which the enzymatic activity can be manipulated by engineered peptide binding. The circular permutant of RNaseT1 was obtained by tethering the original C‐ and N‐termini with a GPAG linker and cleaving the molecule between Glu82 and Asn83. This mutant lacked enzymatic activity, due to the destabilization of entire protein structure. We previously reported the construction of ABC‐type heterotrimeric coiled coil peptides, in which the A‐ and B‐type peptides cannot form the folded trimeric structure without the C‐type peptide. The introduction of the A‐ and B‐type coiled coil peptides to the C‐ and N‐termini of the circular permutant of RNaseT1, respectively, and the subsequent addition of the C‐type coiled coil peptide enabled the RNaseT1 domain to refold properly, thus, restoring the enzymatic activity. The formation of the trimeric coiled coil structure should bring the cleaved sites of RNaseT1 close enough to refold the RNaseT1 domain spontaneously.
Apoptolidinone A: Synthesis of the Apoptolidin A Aglycone
Authors: Schuppan Julia , Wehlan Hermut , Keiper Sonja , Koert Ulrich
Published: July 2006

An efficient stereocontrolled synthesis of apoptolidinone A, the aglycone of apoptolidin A is described. The synthetic strategy relies on a cross coupling between C11/C12 of a northern half (C1–C11) and a southern part (C12–C28) followed by a ring‐size selective macrolactonization. Key steps for the introduction of the southern half stereocenters are a stereoselective aldol reaction, a substrate controlled dihydroxylation and a chelation‐controlled Grignard/aldehyde addition. The conjugated triene of the northern half was built up successively by E‐selective Wittig reactions. L‐Malic acid was chosen as the chiral pool source for the C8/C9 stereocenters. The final cleavage of the silyl ethers and the conversion of the C21 methyl ketal into the hemiketal was achieved by HF⋅pyridine.
Genotoxic Potential of N‐(Benzothiazolyl)sulfonamide Copper(II) Complexes on Yeast Cells Transformed with YEGFP Expression Constructs Containing the RAD54 or RNR2 Promoter
Authors: González‐Álvarez Marta , Alzuet Gloria , del Castillo Lucas , Borrás Joaquín , Liu‐González Malva
Published: July 2006

Four ternary complexes [Cu(L)2(phen)] where L is an N‐(benzothiazol‐2‐yl)sulfonamide derivative have been prepared and their ability to cleave DNA has been studied. The complexes were structurally characterized with the aid of single‐crystal X‐ray crystallography. Whereas the molecular structure of the [Cu(L1)2(phen)] (1) [HL1 = N‐(6‐chlorobenzothiazol‐2‐yl)benzenesulfonamide] and [Cu(L3)2(phen)] (3) [HL3 = N‐(benzothiazol‐2‐yl)benzenesulfonamide] complexes can best be described as having a distorted square‐planar geometry, that of the [Cu(L4)2(phen)] (4) [HL4 = N‐(benzothiazol‐2‐yl)toluenesulfonamide] complex shows a strictly square‐planar geometry. The [Cu(L2)2(phen)MeOH] (2) [HL2 = N‐(6‐chlorobenzothiazol‐2‐yl)toluenesulfonamide] complex displays an axially elongated square‐pyramidal coordination geometry in which the phen ligand binds at the basal plane. Viscosity and fluorescence measurements indicated that [Cu(L4)2(phen)] (4) has a propensity for binding calf thymus DNA. The four complexes were found to be efficient chemical nucleases, with ascorbate/H2O2 activation giving rise to hydroxyl and superoxide radicals as active cleaving species. The nuclease activity of 4 is not only the highest of the four complexes, but also much higher than that of the copper–phenanthroline complex. The ability of the complexes to cleave DNA within cells has been tested by monitoring the expression of the yEGFP gene containing reporter plasmid. The significant induction of fluorescence by complex 4 indicates that it is able to cleave DNA inside the cell. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2006)
Synthesis, Crystal Structures, Cation‐Binding Properties and the Influence of Intramolecular C–H···O Interactions on the Complexation Behaviour of a Family of Cone p‐tert‐Butylcalix[4]arene‐crown‐5 Compounds
Authors: Agnihotri Pragati , Suresh Eringathodi , Paul Parimal , Ghosh Pushpito K.
Published: July 2006

A family of cone p‐tert‐butylcalix[4]arene‐crown‐5 compounds with various substituents (H, COCH3, CH2CO2C2H5 and CH2CO2H) appended at the opposite phenolic oxygen atoms have been synthesised to evaluate their efficiency and selectivity towards different alkali and alkaline‐earth metal ions, and also to ascertain the role of the appended side‐arms in the complexation process. The selectivity of these ionophores towards Na+, K+, Mg2+, and Ca2+ has been evaluated with an aqueous solution containing an equimolar mixture of these ions. The concentration of metal ion in the extract (organic phase) has been estimated by ion chromatographic assay. Among these ions, K+ shows the highest selectivity in all cases except one, where the two phenolic oxygen atoms contain COCH3 substituents. All the ionophores show poor selectivity towards Mg2+ and Ca2+. Association constants (Ka) for the binding of Na+ and K+ to these ionophores have been determined spectrophotometrically. Ka (7.2 × 107) is highest for the binding of K+ to the ionophore with CH2CO2C2H5 substituents. The molecular structures of four of the ionophores and four of the metal complexes have been established by single‐crystal X‐ray crystallography. Analysis of the structures revealed that in case of the ionophore with two COCH3 substituents, the C–H···O interactions form an eight‐membered zigzag ring almost perpendicular to the crown ring, which prevents entry of the metal ions into the calix‐crown cavity. The ionophore with CH2CO2C2H5 substituents, where no such interaction is observed, forms metal complexes easily and exhibits the highest association constant. 1H and 13C NMR studies have also been carried out to investigate the conformational behaviour of these ionophores and their metal complexes in solution.(© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2006)
Structure of the virus capsid protein VP1 of enterovirus 71 predicted by some homology modeling and molecular docking studies
Authors: Ke Yi‐Yu , Chen Yun‐Chu , Lin Thy‐Hou
Published: July 2006

The homology modeling technique has been used to construct the structure of enterovirus 71 (EV 71) capsid protein VP1. The protein is consisted of 297 amino acid residues and treated as the target. The amino acid sequence identity between the target protein and sequences of template proteins 1EAH, 1PIV, and 1D4M searched from NCBI protein BLAST and WorkBench protein tools were 38, 37, and 36%, respectively. Based on these template structures, the protein model was constructed by using the InsightII/Homology program. The protein model was briefly refined by energy minimization and molecular dynamics (MD) simulation steps. The protein model was validated using some web available servers such as ERRAT, PROCHECK, PROVE, and PROSA2003. However, an inconsistency between the docking scores and the measured activity was observed for a series of EV 71 VP1 inhibitors synthesized by Shia et al. (J Med Chem 2002, 45, 1644) and docked into the binding pocket of the protein model using the DOCK 4.0.2 program. The protein model with an EV 71 VP1 inhibitor docked and engulfed was then refined further by some MD simulation steps in the presence of water molecules. The docking scores obtained for these inhibitors after such a MD refinement were well correlated with the activities. The structure–activity relationships for the ligand–protein model system was also analyzed using the GRID‐VOLSURF programs and the corresponding noncrossvalidated and crossvalidated (by leave‐one‐out) r2 and q2 were 0.99 and 0.61, respectively. The hydrophobic nature of the binding pocket of the protein model was also examined using the GRID21 program. The possibility of improving the potency of the current series of EV 71 VP1 inhibitors was discussed based on all the studies presented. © 2006 Wiley Periodicals, Inc. J Comput Chem 27: 1556–1570, 2006
Ultrasound findings of teres minor denervation in suspected quadrilateral space syndrome
Authors: Brestas Paraskevas S. , Tsouroulas Marinos , Nikolakopoulou Zoi , Malagari Katerina , Drossos Charalampos
Published: July 2006

Isolated teres minor denervation is an uncommon finding on sonographic examination. We present a case of a 64‐year‐old man with increased echogenity of the teres minor muscle and a slight reduction in muscle bulk. Investigation of a suspected axillary nerve lesion included a detailed sonographic examination of the posterior shoulder and the axillary space, followed by MR imaging and electrophysiologic testing. This case demonstrates the potential importance of examining rotator cuff muscles when performing sonographic examination of the shoulder in patients with persistent symptoms, no history of trauma, and absence of tendon tears. © 2006 Wiley Periodicals, Inc. J Clin Ultrasound 34:343–347, 2006
Relationship of increased renal cortical echogenicity with clinical and laboratory findings in pediatric renal disease
Authors: Kasap Belde , Soylu Alper , Türkmen Mehmet , Kavukcu Salih
Published: July 2006

Purpose.

To correlate the clinical and laboratory findings with increased renal cortical echogenicity in children with acute renal diseases.

Materials and Methods.

Children with increased renal cortical echogenicity were evaluated retrospectively. Laboratory findings and final diagnoses were reviewed. Patients with increased echogenity were classified into group 1 (mild; grade I) or group 2 (more severe; grades II and III).

Results.

There were 7 newborns and 114 children (67 male, 47 female) with increased renal cortical echogenicity with a mean (±SD) age of 7.0 (±4.4) years. The clinical diagnosis was anatomic abnormality (including vesicoureteral reflux, ureteropelvic junction obstruction, ureterovesical junction obstruction, double collecting system) in 9%, urinary tract infection in 21%, urolithiasis in 6%, nephrotic syndrome in 20%, glomerulonephritis in 32%, and other diseases in 12%. Hyperechogenicity was bilateral in 72%, right‐sided in 19%, and left‐sided in 9%. There were 81 patients in group 1 and 33 patients in group 2 (grade II, 29; grade III, 4). There was no statistically significant difference between the groups with regard to age, sex, and serum blood urea nitrogen level, serum creatinine level, uric acid level, urine pH, and specific gravity. Hematuria was more frequent in group 2, whereas proteinuria and pyuria incidences were similar in the 2 groups. In patients with hematuria, glomerulonephritis was the most common cause.

Conclusions.

Glomerulonephritis is the most frequent acute disease causing increased renal echogenicity in childhood, and higher echogenicity is more likely to be associated with hematuria. © 2006 Wiley Periodicals, Inc. J Clin Ultrasound 34:339–342, 2006
Calcium‐regulated fusion of yolk granules during early embryogenesis of Periplaneta americana
Authors: Ramos I.B. , Miranda K. , de Souza W. , Machado E.A.
Published: July 2006

This work reported membrane fusion of yolk granules (YGs) during early embryogenesis of the insect Periplaneta americana (P. americana). We showed that eggs from Day 5 of embryogenesis possess a greater amount of enlarged YGs in comparison with Day 1. Day 5 is also the period when the largest amount of free calcium is found (∼17 mM) within the oothecae from early embryogenesis. Treatment of Day 1‐YGs fraction with 17 mM Ca2+ resulted in a YG size pattern very similar to the one observed in Day 5 eggs, where enlarged YGs were formed. YG membrane fusion was observed by fluorescent membrane dye transfer from previously labeled small YGs to larger ones and was also visualized by electron microscopy. We also showed that the small “in fusion” YGs seemed to be acidic, suggesting that acidification is correlated with YG membrane fusion. Hence, it was shown that YGs are capable of membrane fusion in a calcium‐dependent manner and this process probably occurs in vivo during early embryogenesis of P. americana. Mol. Reprod. Dev. © 2006 Wiley‐Liss, Inc.
Comparative analysis of apoptotic pathways in rat, mouse, and hamster spermatozoa
Authors: Cisternas Pablo , Moreno Ricardo D.
Published: July 2006

Apoptosis is a type of cell death characterized by the activation of a family of cysteine‐proteases called caspases. We made a comparative study to determine the presence of several caspases and other regulators of apoptosis in rat, mouse, and hamster spermatozoa. Our results showed that the three species have both active and inactive caspases‐8 and ‐3, the proapoptotic protein BID, p53, and the endogenous caspase inhibitor cIAP‐1. However, we did not find evidence for the presence of active caspase‐9. The acrosome reaction (i.e., the exocytic process of sperm acrosome) and sperm viability were not affected by the presence of a general caspase inhibitor. On the other hand, valinomycin, which promotes caspase‐dependent cell death in somatic cells, induced caspase‐independent cell death in spermatozoa. TRAIL, a ligand whose receptor induces apoptosis in malignant cells, did not have any effect in the viability of mouse spermatozoa, despise the presence of its receptor in rat and mouse, but not in hamster spermatozoa. Therefore, our results strongly suggest that rodent spermatozoa have some components of the apoptotic pathway. However, the role of caspases in mammalian spermatozoa appears to be unrelated to sperm survival or to the acrosome reaction under physiological conditions. Mol. Reprod. Dev. © 2006 Wiley‐Liss, Inc.
Government engagement with non‐state providers of water and sanitation services
Authors: Sansom Kevin
Published: July 2006

Increasingly, governments in developing countries recognise that the public sector alone cannot provide adequate water and sanitation services to all. Non‐state providers (NSPs) including both formal and informal private providers, as well as civil society institutions, also have important roles to play. There are clear challenges for governments intending to work with NSPs, not least of which is the institutional compatibility between bureaucratic agencies and informal water and sanitation NSPs. However, positive examples of government agencies working effectively with NSPs are emerging in many countries. Government engagement with water and sanitation NSPs can be split into five main types: recognition, dialogue, facilitation/collaboration, contracting and regulation. In many situations, a lack of formal recognition of water or sanitation NSPs is an impediment to more productive forms of engagement. There are a number of potential intervention options within each of the five types of engagement that government agencies should carefully consider when supporting the development of NSP water and sanitation services. Copyright © 2006 John Wiley & Sons, Ltd.
The effect of silver doping on the critical current density of Bi‐Pb‐Sr‐Ca‐Cu‐O ceramic superconductor
Authors: Zargar Shoushtari M. , Bahrami Amir , Farbod Mansoor
Published: July 2006

In this research, the effect of silver doping on Bi1.6Pb0.4Sr2Ca2Cu3Oy ceramic superconductor has been investigated. The solid‐state reaction method and two different silver doping methods has been used, namely, doping during making processes of samples (batch 1) and doping after making Bi1.6Pb0.4Sr2Ca2Cu3Oy superconductor (batch 2). We observed that by adding silver to BPSCCO compound, partial melting temperature of the compound is decreased. The critical current density (Jc) in both batches is affected by Ag doping. The investigation of SEM images of samples has shown that the all surfaces of the samples are porous and the grains are plate like. It seems that the BPSCCO grains in batch 1 samples are coated with silver but in the samples of batch 2, the silver also sits between the BPSCCO grains. The XRD patterns studies indicated that the silver peaks form separate phase and also by adding silver to BSCCO, the BPSCCO peaks do not show considerable shift. (© 2006 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Design of ac susceptometer using closed cycle helium cryostat
Authors: Shahabuddin Mohammed , Alzayed Nasser Saleh
Published: July 2006

The design of the low cost ac susceptometer using closed cycle helium cryostat (CCHC) having He gas exchange sample well is discussed. The problem of higher frequency limitation generally in CCHC has been solved. The sample temperature is varied independently, keeping the coil system at the lowest temperature achieved in the sample well of the CCHC. This increases the bridge sensitivity. The nullification of the offset voltage up to 100 nV is achieved simply sliding the secondary along the primary. The whole systems can be inserted and taken out through the gate valve of the sample well of the CCHC so that it dose not affect the other sample holders for different measurement. (© 2006 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Simulation of YBa2Cu3O7/MgO surface growth
Authors: Mohammadizadeh M. R. , Safari N. , Kia M. , Savaloni H.
Published: July 2006

For surface growth simulation of YBa2Cu3O7on MgO substrate, binding energies between each two different Y, Ba, Cu, O, and Mg atoms were calculated byab initiopseudopotential density functional theory approach. Then, simulation of YBa2Cu3O7growth was performed by a simple two dimensional model based on the ballistic aggregation of hard discs. By increasing the substrate temperature, the atomic layers distribution is more condensed and the nanometric surface roughness decreases. (© 2006 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Resistivity anisotropy of the Bi‐based misfit layer compounds (BiS)1.11(NbS2) and (BiSe)1.10(NbSe2) down to the liquid nitrogen temperature
Authors: Nader A.
Published: July 2006

The resistivity in the transverse and parallel directions of the superconducting misfit layer compounds (BiS)1.11(NbS2) and (BiSe)1.10(NbSe2) has been measured down to the liquid nitrogen temperature. The resistivity anisotropy for these two compounds was found to increase when the temperature is lowered and its value in the temperature range of our measurement is two orders of magnitude greater than the value expected by Lawrence and Doniach model. (© 2006 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Diagnosis of feline leukaemia virus infection by semi-quantitative real-time polymerase chain reaction
Authors: Pinches Mark D.G. , Helps Christopher R. , Gruffydd-Jones Tim J. , Egan Kathy , Jarrett Oswald , Tasker Séverine
Published: July 2006

In this paper the design and use of a semi-quantitative real-time polymerase chain reaction assay (RT-PCR) for feline leukaemia virus (FeLV) provirus is described. Its performance is evaluated against established methods of FeLV diagnosis, including virus isolation and enzyme-linked immunoassay (ELISA) in a population of naturally infected cats. The RT-PCR assay is found to have both a high sensitivity (0.92) and specificity (0.99) when examined by expectation maximisation methods and is also able to detect a large number of cats with low FeLV proviral loads that were negative by other conventional test methods.