bluuebunny/crossref_metadata_2025 huggingface dataset:
Enhanced Tolerance to Environmental Stress in Transgenic Plants Expressing the Transcriptional Coactivator Multiprotein Bridging Factor 1c
Authors: Suzuki Nobuhiro , Rizhsky Ludmila , Liang Hongjian , Shuman Joel , Shulaev Vladimir , Mittler Ron
Published: October 2005

Abiotic stresses cause extensive losses to agricultural production worldwide. Acclimation of plants to abiotic conditions such as drought, salinity, or heat is mediated by a complex network of transcription factors and other regulatory genes that control multiple defense enzymes, proteins, and pathways. Associated with the activity of different transcription factors are transcriptional coactivators that enhance their binding to the basal transcription machinery. Although the importance of stress-response transcription factors was demonstrated in transgenic plants, little is known about the function of transcriptional coactivators associated with abiotic stresses. Here, we report that constitutive expression of the stress-response transcriptional coactivator multiprotein bridging factor 1c (MBF1c) in Arabidopsis (Arabidopsis thaliana) enhances the tolerance of transgenic plants to bacterial infection, heat, and osmotic stress. Moreover, the enhanced tolerance of transgenic plants to osmotic and heat stress was maintained even when these two stresses were combined. The expression of MBF1c in transgenic plants augmented the accumulation of a number of defense transcripts in response to heat stress. Transcriptome profiling and inhibitor studies suggest that MBF1c expression enhances the tolerance of transgenic plants to heat and osmotic stress by partially activating, or perturbing, the ethylene-response signal transduction pathway. Present findings suggest that MBF1 proteins could be used to enhance the tolerance of plants to different abiotic stresses.
Three Maize Leaf Ferredoxin:NADPH Oxidoreductases Vary in Subchloroplast Location, Expression, and Interaction with Ferredoxin
Authors: Okutani Satoshi , Hanke Guy T. , Satomi Yoshinori , Takao Toshifumi , Kurisu Genji , Suzuki Akira , Hase Toshiharu
Published: October 2005

In higher plants, ferredoxin (Fd):NADPH oxidoreductase (FNR) catalyzes reduction of NADP+ in the final step of linear photosynthetic electron transport and is also implicated in cyclic electron flow. We have identified three leaf FNR isoenzymes (LFNR1, LFNR2, and LFNR3) in maize (Zea mays) chloroplasts at approximately equivalent concentrations. Fractionation of chloroplasts showed that, while LFNR3 is an exclusively soluble enzyme, LFNR1 is only found at the thylakoid membrane and LFNR2 has a dual location. LFNR1 and LFNR2 were found to associate with the cytochrome b6f complex following its partial purification. We cloned LFNR3 and produced all three isoenzymes as stable, soluble proteins. Measurement of Fd reduction ability showed no significant differences between these recombinant enzymes. Column chromatography revealed variation between the interaction mechanisms of LFNR1 and LFNR2 with Fd, as detected by differential dependence on specific intermolecular salt bridges and variable sensitivity of interactions to changes in pH. A comparison of LFNR transcripts in leaves of plants grown on variable nitrogen regimes revealed that LFNR1 and LFNR2 transcripts are relatively more abundant under conditions of high demand for NADPH. These results are discussed in terms of the functional differentiation of maize LFNR isoenzymes.
TRANSPARENT TESTA10 Encodes a Laccase-Like Enzyme Involved in Oxidative Polymerization of Flavonoids in Arabidopsis Seed Coat
Authors: Pourcel Lucille , Routaboul Jean-Marc , Kerhoas Lucien , Caboche Michel , Lepiniec Loïc , Debeaujon Isabelle
Published: October 2005

The Arabidopsis thaliana  transparent testa10 (tt10) mutant exhibits a delay in developmentally determined browning of the seed coat, also called the testa. Seed coat browning is caused by the oxidation of flavonoids, particularly proanthocyanidins, which are polymers of flavan-3-ol subunits such as epicatechin and catechin. The tt10 mutant seeds accumulate more epicatechin monomers and more soluble proanthocyanidins than wild-type seeds. Moreover, intact testa cells of tt10 cannot trigger H2O2-independent browning in the presence of epicatechin and catechin, in contrast with wild-type cells. UV–visible light detection and mass spectrometry revealed that the major oxidation products obtained with epicatechin alone are yellow dimers called dehydrodiepicatechin A. These products differ from proanthocyanidins in the nature and position of their interflavan linkages. Flavonol composition was also affected in tt10 seeds, which exhibited a higher ratio of quercetin rhamnoside monomers versus dimers than wild-type seeds. We identified the TT10 gene by a candidate gene approach. TT10 encodes a protein with strong similarity to laccase-like polyphenol oxidases. It is expressed essentially in developing testa, where it colocalizes with the flavonoid end products proanthocyanidins and flavonols. Together, these data establish that TT10 is involved in the oxidative polymerization of flavonoids and functions as a laccase-type flavonoid oxidase.
Functional Differentiation of Bundle Sheath and Mesophyll Maize Chloroplasts Determined by Comparative Proteomics
Authors: Majeran Wojciech , Cai Yang , Sun Qi , van Wijk Klaas J.
Published: October 2005

Chloroplasts of maize (Zea mays) leaves differentiate into specific bundle sheath (BS) and mesophyll (M) types to accommodate C4 photosynthesis. Consequences for other plastid functions are not well understood but are addressed here through a quantitative comparative proteome analysis of purified M and BS chloroplast stroma. Three independent techniques were used, including cleavable stable isotope coded affinity tags. Enzymes involved in lipid biosynthesis, nitrogen import, and tetrapyrrole and isoprenoid biosynthesis are preferentially located in the M chloroplasts. By contrast, enzymes involved in starch synthesis and sulfur import preferentially accumulate in BS chloroplasts. The different soluble antioxidative systems, in particular peroxiredoxins, accumulate at higher levels in M chloroplasts. We also observed differential accumulation of proteins involved in expression of plastid-encoded proteins (e.g., EF-Tu, EF-G, and mRNA binding proteins) and thylakoid formation (VIPP1), whereas others were equally distributed. Enzymes related to the C4 shuttle, the carboxylation and regeneration phase of the Calvin cycle, and several regulators (e.g., CP12) distributed as expected. However, enzymes involved in triose phosphate reduction and triose phosphate isomerase are primarily located in the M chloroplasts, indicating that the M-localized triose phosphate shuttle should be viewed as part of the BS-localized Calvin cycle, rather than a parallel pathway.
PGP4, an ATP Binding Cassette P-Glycoprotein, Catalyzes Auxin Transport in Arabidopsis thaliana Roots
Authors: Terasaka Kazuyoshi , Blakeslee Joshua J. , Titapiwatanakun Boosaree , Peer Wendy A. , Bandyopadhyay Anindita , Makam Srinivas N. , Lee Ok Ran , Richards Elizabeth L. , Murphy Angus S. , Sato Fumihiko , Yazaki Kazufumi
Published: October 2005

Members of the ABC (for ATP binding cassette) superfamily of integral membrane transporters function in cellular detoxification, cell-to-cell signaling, and channel regulation. More recently, members of the multidrug resistance P-glycoprotein (MDR/PGP) subfamily of ABC transporters have been shown to function in the transport of the phytohormone auxin in both monocots and dicots. Here, we report that the Arabidopsis thaliana MDR/PGP PGP4 functions in the basipetal redirection of auxin from the root tip. Reporter gene studies showed that PGP4 was strongly expressed in root cap and epidermal cells. PGP4 exhibits apolar plasma membrane localization in the root cap and polar localization in tissues above. Root gravitropic bending and elongation as well as lateral root formation were reduced in pgp4 mutants compared with the wild type. pgp4 exhibited reduced basipetal auxin transport in roots and a small decrease in shoot-to-root transport consistent with a partial loss of the redirective auxin sink in the root. Seedlings overexpressing PGP4 exhibited increased shoot-to-root auxin transport. Heterologous expression of PGP4 in mammalian cells resulted in 1-N-naphthylthalamic acid–reversible net uptake of [3H]indole-3-acetic acid. These results indicate that PGP4 functions primarily in the uptake of redirected or newly synthesized auxin in epidermal root cells.
Transcriptome and Selected Metabolite Analyses Reveal Multiple Points of Ethylene Control during Tomato Fruit Development
Authors: Alba Rob , Payton Paxton , Fei Zhanjun , McQuinn Ryan , Debbie Paul , Martin Gregory B. , Tanksley Steven D. , Giovannoni James J.
Published: October 2005

Transcriptome profiling via cDNA microarray analysis identified 869 genes that are differentially expressed in developing tomato (Solanum lycopersicum) pericarp. Parallel phenotypic and targeted metabolite comparisons were employed to inform the expression analysis. Transcript accumulation in tomato fruit was observed to be extensively coordinated and often completely dependent on ethylene. Mutation of an ethylene receptor (Never-ripe [Nr]), which reduces ethylene sensitivity and inhibits ripening, alters the expression of 37% of these 869 genes. Nr also influences fruit morphology, seed number, ascorbate accumulation, carotenoid biosynthesis, ethylene evolution, and the expression of many genes during fruit maturation, indicating that ethylene governs multiple aspects of development both prior to and during fruit ripening in tomato. Of the 869 genes identified, 628 share homology (E-value ≤1 × 10−10) with known gene products or known protein domains. Of these 628 loci, 72 share homology with previously described signal transduction or transcription factors, suggesting complex regulatory control. These results demonstrate multiple points of ethylene regulatory control during tomato fruit development and provide new insights into the molecular basis of ethylene-mediated ripening.
A novel representation of protein sequences for prediction of subcellular location using support vector machines
Authors: Matsuda Setsuro , Vert Jean‐Philippe , Saigo Hiroto , Ueda Nobuhisa , Toh Hiroyuki , Akutsu Tatsuya
Published: October 2005

As the number of complete genomes rapidly increases, accurate methods to automatically predict the subcellular location of proteins are increasingly useful to help their functional annotation. In order to improve the predictive accuracy of the many prediction methods developed to date, a novel representation of protein sequences is proposed. This representation involves local compositions of amino acids and twin amino acids, and local frequencies of distance between successive (basic, hydrophobic, and other) amino acids. For calculating the local features, each sequence is split into three parts: N‐terminal, middle, and C‐terminal. The N‐terminal part is further divided into four regions to consider ambiguity in the length and position of signal sequences. We tested this representation with support vector machines on two data sets extracted from the SWISS‐PROT database. Through fivefold cross‐validation tests, overall accuracies of more than 87% and 91% were obtained for eukaryotic and prokaryotic proteins, respectively. It is concluded that considering the respective features in the N‐terminal, middle, and C‐terminal parts is helpful to predict the subcellular location.
Hydrogen‐bonded turns in proteins: The case for a recount
Authors: Panasik Nick , Fleming Patrick J. , Rose George D.
Published: October 2005

β‐Turns are sites at which proteins change their overall chain direction, and they occur with high frequency in globular proteins. The Protein Data Bank has many instances of conformations that resemble β‐turns but lack the characteristic N–H(i) → O=C(i − 3) hydrogen bond of an authentic β‐turn. Here, we identify potential hydrogen‐bonded β‐turns in the coil library, a Web‐accessible database utility comprised of all residues not in repetitive secondary structure, neither α‐helix nor β‐sheet (http://www.roselab.jhu.edu/coil). In particular, candidate turns were identified as four‐residue segments satisfying highly relaxed geometric criteria but lacking a strictly defined hydrogen bond. Such candidates were then subjected to a minimization protocol to determine whether slight changes in torsion angles are sufficient to shift the conformation into reference‐quality geometry without deviating significantly from the original structure. This approach of applying constrained minimization to known structures reveals a substantial population of previously unidentified, stringently defined, hydrogen‐bonded β‐turns. In particular, 33% of coil library residues were classified as β‐turns prior to minimization. After minimization, 45% of such residues could be classified as β‐turns, with another 8% in 310 helixes (which closely resemble type III β‐turns). Of the remaining coil library residues, 37% have backbone dihedral angles in left‐handed polyproline II structure.
Undistorted structural analysis of soluble proteins by attenuated total reflectance infrared spectroscopy
Authors: Goldberg Michel E. , Chaffotte Alain F.
Published: October 2005

Water from the solvent very strongly absorbs light in the frequency range of interest for studying protein structure by infrared (IR) spectroscopy. This renders handling of the observation cells painstaking and time consuming, and limits the reproducibility of the measurements when IR spectroscopy is applied to proteins in aqueous solutions. These difficulties are circumvented by the use of an Attenuated Total Reflectance (ATR) accessory. However, when protein solutions are studied, ATR spectroscopy suffers from several drawbacks, the most severe being nonproportionality of the observed absorbance with the protein concentration and spectral distortions that vary from protein to protein and from sample to sample. In this study, we show (1) that the nonproportionality is due to adsorption of the protein on the ATR crystal surface; (2) that the contribution of the crystal‐adsorbed protein can easily be taken into account, rendering the corrected absorbance proportional to the protein concentration; (3) that the observed variable base line distortions, likely due to changes in the penetration depth of the light beam in solutions with the refractive index that depends on the protein concentration, can be easily eliminated; and (4) that ATR IR spectra thus corrected for protein adsorption and light penetration can be used to properly analyze the secondary structure of proteins in solution.
Insights into the role of the metal binding site in methionine‐R‐sulfoxide reductases B
Authors: Olry Alexandre , Boschi‐Muller Sandrine , Yu Hong , Burnel Daniel , Branlant Guy
Published: October 2005

Methionine sulfoxide reductases B (MsrBs) catalyze the reduction of methionine‐R‐sulfoxide via a three‐step chemical mechanism including a reductase step, formation of an intradisulfide bond followed by a thioredoxin recycling process. Fifty percent of the MsrBs, including the Escherichia coli enzyme, possess a metal binding site composed of two CXXC motifs of unknown function. It is located on the opposite side of the active site. The overexpressed E. coli MsrB tightly binds one atom of zinc/iron. Substitution of the cysteines of E. coli MsrB results in complete loss of bound metal and reductase activity, and leads to a low‐structured conformation of the protein as shown by CD, fluorescence, and DSC experiments. Introduction of the two CXXC motifs in Neisseria meningitidis MsrB domain leads to a MsrB that tightly binds one atom of zinc/iron, shows a strongly increased thermal stability and displays a reductase activity similar to that of the wild‐type but lacking thioredoxin recycling activity. These results demonstrate the stabilizing effect of the metal and the existence of a preformed metal binding site in the nonbound metal MsrB. The data also indicate that metal binding to N. meningitidis MsrB induces subtle structural modifications, which prevent formation of a competent binary complex between oxidized MsrB and reduced thioredoxin but not between reduced MsrB and substrate. The fact that the E. coli and the N. meningitidis MsrBs exhibit a similar thermal stability suggests the existence of other structural factors in the nonbound metal MsrBs that compensate the metal bound stabilizing effect.
Serum biomarkers for cardiovascular inflammation in subclinical hypothyroidism
Authors: Hueston William J. , King Dana E. , Geesey Mark E.
Published: October 2005

Objective Cardiovascular disease has been reported to be more common in patients with elevated levels of TSH and with normal thyroxine levels, termed subclinical hypothyroidism (SCH). The aim of this study was to determine whether individuals with SCH were more likely than euthyroid controls to have elevations in inflammatory biomarkers that are associated with cardiac disease.

Methods From the National Health and Nutrition Examination Survey (NHANES) 1999–2002 data we identified respondents aged ≥ 40 who met the laboratory criteria for SCH. We compared the average values and frequencies of elevated C‐reactive protein (CRP) and homocysteine in the SCH individuals to those in euthyroid individuals.

Results A total of 1608 individuals were included in the analysis of whom 45 (2·8%) met the criteria for SCH, with seven (15·6%) having TSH levels between 10 and 15 IU/l. We found no differences in median, mean or the percentage of elevated CRP or homocysteine in individuals with SCH compared to euthyroid controls. Linear regression models adjusting for age, gender, the presence of other cardiac risk factors, and whether the individual was using a cholesterol‐lowering drug also failed to show any association between SCH and either high‐sensitivity CRP (hsCRP) (P = 0·62) or homocysteine (P = 0·63) levels. When we examined the subset of people with TSH levels between 10 and 15 IU/l, we again found no difference in mean hsCRP or homocysteine compared to control.

Conclusions In this large population‐based sample, hsCRP and homocysteine levels do not differ for individuals with SCH compared to euthyroid individuals. This suggests that cardiac inflammation is not greater in individuals with SCH.
The superoxide‐producing NAD(P)H oxidase Nox4 in the nucleus of human vascular endothelial cells
Authors: Kuroda Junya , Nakagawa Kazunori , Yamasaki Tomoko , Nakamura Kei‐ichiro , Takeya Ryu , Kuribayashi Futoshi , Imajoh‐Ohmi Shinobu , Igarashi Kazuhiko , Shibata Yosaburo , Sueishi Katsuo , Sumimoto Hideki
Published: October 2005

The superoxide‐producing NAD(P)H oxidase Nox4 was initially identified as an enzyme that is highly expressed in the kidney and is possibly involved in oxygen sensing and cellular senescence. Although the oxidase is also abundant in vascular endothelial cells, its role remains to be elucidated. Here we show that Nox4 preferentially localizes to the nucleus of human umbilical vein endothelial cells (HUVECs), by immunocytochemistry and immunoelectron microscopy using three kinds of affinity‐purified antibodies raised against distinct immunogens from human Nox4. Silencing of Nox4 by RNA interference (RNAi) abrogates nuclear signals given with the antibodies, confirming the nuclear localization of Nox4. The nuclear fraction of HUVECs exhibits an NAD(P)H‐dependent superoxide‐producing activity in a manner dependent on Nox4, which activity can be enhanced upon cell stimulation with phorbol 12‐myristate 13‐acetate. This stimulant also facilitates gene expression as estimated in the present transfection assay of HUVECs using a reporter regulated by the Maf‐recognition element MARE, a DNA sequence that constitutes a part of oxidative stress response. Both basal and stimulated transcriptional activities are impaired by RNAi‐mediated Nox4 silencing. Thus Nox4 appears to produce superoxide in the nucleus of HUVECs, thereby regulating gene expression via a mechanism for oxidative stress response.
Viability assessment of lactic acid bacteria in commercial dairy products stored at 4 °C using LIVE/DEAD® BacLightTM staining and conventional plate counts
Authors: Moreno Yolanda , Collado M. Carmen , Ferrús M. Antonia , Cobo José M. , Hernández Enrique , Hernández Manuel
Published: October 2005

The viability of lactic acid bacteria (LAB) from commercial fermented milks was studied during storage at 4 °C. The enumeration of total viable bacteria was assessed using fluorescence microscopy. Plate counting on selective media was used to enumerate LAB. Using LIVE/DEAD® BacLightTM viability staining, it was observed that bacterial counts decreased gradually after expiry dates, the number of viable bacteria remaining above 106 bacteria g−1 for all of the products. Viable cell counts estimated by plating onto selective media were lower than those obtained by direct microscopic counting. The viability of LAB contained in acid products decreased during their storage period at 4 °C. All products contain viable LAB ranging from 108 to 109 bacteria g−1 and could be considered as probiotic, given that the recommended minimum number of probiotic bacteria in such food products is approximately 107 bacteria mL−1 product. The number of bifidobacteria in commercial fermented milks declared to contain bifidobacteria varied from 104 to 107 bacteria mL−1. This study confirms the usefulness of fluorescent techniques for a rapid and accurate evaluation of bacterial viability in probiotic products.
Chemical and microbiological quality of Garris, Sudanese fermented camel's milk product
Authors: Sulieman Abdel Moneim El‐Hadi , Ilayan Abdalla Adam , Faki Ahmed El‐Awad El
Published: October 2005

In the present study, some of the chemical and microbiological characteristics of garris, a Sudanese traditionally fermented camel's milk product, were investigated. The chemical analyses included, pH, titrable acidity and ethanol contents. A total of 100 strains of lactic acid bacteria (LAB) were isolated from twenty samples of traditionally fermented household garris. The selected isolates were phenotypically characterized by their ability to ferment 49 carbohydrates using API 50 CHL kits and additional biochemical tests. LAB dominated the microflora of garris samples, and the major genera were Lactobacillus (74%), followed by Lactococcus (12%), Enterococcus (10%) and Leuconostocs (4%). The most predominant Lactobacillus species were identified as Lactobacillus paracasei ssp. paracasei (64 strains), L. fermentum (seven strains) and only three strains as L. plantarum. Most strains produced the enzymes that are relevant to cultured dairy product processing. The Lactococcus species were identified as Lactococcus lactis. The average pH value of the samples was 4.42 ± 0.21. The pH values were accompanied with increasing of titrable acidity which averaged 1.72 ± 0.04%. The relatively high amounts of ethanol detected in all samples (average 1.40 ± 0.03%) together with the high yeasts counts (6.0 ± 0.53 log10 cfu mL−1), indicated that the fermentation process of garris is a yeast‐lactic fermentation.
Composition of dishes consumed in Cameroon
Authors: Ponka Roger , Fokou Elie , Fotso Martin , Tchouanguep Félicité Mbiapo , Leke Rose , Souopgui Jacob , Bih Mercy Achu
Published: October 2005

This study deals with determination of some of the mineral salts and phytate contents in the food consumed by inhabitants of Cameroon known as Ngali II, a rural area, which is a malaria endemic zone. The food consumed by the people of this area are light and thick sauces prepared from several green leafy vegetables, leguminous seeds and cucurbit seeds (egusi seeds), pastes prepared from leguminous seeds or egusi seeds complemented with different tubers and plantains. The mineral salt contents were determined by atomic absorption spectrophotometry and phytate by spectrophotometric methods. The results are expressed in mg (100 g)−1 dry weight. Iron: 1.32–28.46, zinc: 2.0–7.28, copper: 0.13–1.09, magnesium: 24.5–369.8 and phytate: 9.08–1988.2. This study reveals that a high consumption of food rich in zinc will greatly reduce the high prevalence of malaria in Ngali II.
Ethnic differences in the presentation of chronic hepatitis C
Authors: Lepe R. , Layden‐Almer J. E. , Layden T. J. , Cotler S.
Published: October 2005

Summary.  Hispanics comprise 13% of the population in the United States and are the fastest growing minority group. Features of hepatitis C in Hispanics have not been well characterized. The aims of this study were to compare features of hepatitis C among Hispanics, Whites, and African–Americans and to characterize hepatitis C infection in Hispanics. A retrospective analysis was performed on 1225 consecutive patients with hepatitis C seen at the University of Illinois at Chicago including 227 Hispanics, 508 Whites, and 490 African–Americans. Data collection consisted of demographic variables, risk factors for hepatitis C, history of alcohol use, laboratory parameters and liver histology. Pair‐wise comparisons showed that Hispanics had higher aminotransferase levels than Whites and African–Americans. Hispanics had higher portal inflammation scores on liver biopsy than African–Americans (P = 0.002) and Whites (P = 0.043). Hispanics had a higher frequency of cirrhosis than African–Americans (P < 0.001) and a trend towards more cirrhosis than Whites (P = 0.165). There was a trend towards a higher prevalence of cirrhosis in Hispanic women (56%) than in Hispanic men (45%) [P = 0.14]. A cross‐sectional analysis of patients at our liver center showed that Hispanics with hepatitis C had higher aminotransferase levels, more portal inflammation than Whites and African–Americans, and a higher prevalence of cirrhosis than African–Americans.
Efficacy of repeated high‐dose hepatitis B vaccine (80 μg) in patients with chronic liver disease
Authors: Aziz A. , Aziz S. , Li D. S. , Murphy L. , Leone N. , Kennedy M. , Dhillon S. , Van Thiel D. H.
Published: October 2005

Summary.  Patients with chronic liver disease (CLD) respond poorly to standard hepatitis B (HBV) vaccine given as sequential 20 μg IM shots because of an overall impaired immune response. Many of these patients go on to liver transplantation and are at risk of acquiring recurrent or de novo HBV infection.

To evaluate the efficacy and safety of high‐dose (80 μg) IM HBV vaccination in patients with CLD who had previously failed to respond to a standard three‐dose schedule of 40 μg IM vaccine given monthly.

A retrospective review was undertaken at our institution of 79 patients with CLD who were treated with high‐dose (80 μg) HBV vaccinations. All had previously failed a three‐dose course of 40 μg HBV vaccine. An HBV vaccine response was defined as an anti‐HBs titer greater than 100 mIU/ml. Liver enzymes, creatinine, age, prothrombin time, total vaccine dose, and MELD score were recorded.

No adverse events were reported. Seventy‐two per cent (57/79) of the subjects had an adequate response after receiving a mean total dose of 220 μg vaccine (range 80–800 μg). Twenty‐eight per cent (22/79) of the subjects did not respond after receiving a mean total dose of 420 μg vaccine (range 240–720 μg). Non‐responders had more severe hepatic disease defined as a higher mean total bilirubin level (p = 0.003) and a lower mean albumin level (p < 0.05). Age, prothrombin time, MELD score, and creatinine were not statistically significant between the responders and non‐responders.

Repeated high‐dose (80 μg) HBV vaccination, in patients who do not respond to standard HBV vaccine doses, is safe and effective in the majority of patients with CLD.
Diabetes mellitus in chronic hepatitis B and C: prevalence and potential association with the extent of liver fibrosis
Authors: Papatheodoridis G. V. , Chrysanthos N. , Savvas S. , Sevastianos V. , Kafiri G. , Petraki K. , Manesis E. K.
Published: October 2005

Summary.  Diabetes mellitus has been reported to have an increased prevalence and to be associated with more severe fibrosis in patients with chronic hepatitis C. We evaluated the prevalence of diabetes mellitus in patients with chronic hepatitis B or C as well as the possible association between presence of diabetes and extent of liver fibrosis. In total, 434 consecutive patients with histologically documented hepatitis B e antigen (HBeAg)‐negative chronic hepatitis B (n = 174) or chronic hepatitis C (n = 260) were studied. The relationships of diabetes and epidemiological, somatomorphic, laboratory and histological patient characteristics were evaluated. Liver histological lesions were blindly evaluated according to the Ishak's classification. Diabetes was present in 58 (13%) patients, without any difference between those with chronic hepatitis B (14%) or C (13%). Diabetes was observed significantly less frequently in patients with fibrosis score 0–2 (7.7%) than 3–4 (10.4%) than 5–6 (29.2%) (P < 0.001). The presence of diabetes was independently associated with higher gamma‐glutamyl‐transpeptidase (GGT) levels and more severe fibrosis or presence of cirrhosis (P < 0.001) as well as with presence of hepatic steatosis and increased serum triglycerides levels (P < 0.02). In the noncirrhotic patients, diabetes was significantly associated with older age and higher GGT levels, but not with the extent of fibrosis. In conclusion, diabetes mellitus is observed in more than 10% of patients with either HBeAg‐negative chronic hepatitis B or chronic hepatitis C. The presence of diabetes is strongly associated with more severe liver fibrosis, but such an association may be related to the high prevalence of diabetes in patients with cirrhosis.
Modulation of the anti‐inflammatory interleukin 10 and of proapoptotic IL‐18 in patients with chronic hepatitis C treated with interferon alpha and ribavirin
Authors: Marín‐Serrano E. , Rodríguez‐Ramos C. , Díaz F. , Martín‐Herrera L. , Girón‐González J. A.
Published: October 2005

Summary.  The aim of this work was to analyse apoptosis rate, measured by the serum levels of proapoptotic interleukin (IL)‐18 and of soluble Fas (sFas), as well as of anti‐inflammatory IL‐10, in patients with chronic hepatitis C, at baseline and after treatment with interferon alpha and ribavirin. Twenty‐seven patients with biopsy‐proven chronic hepatitis C were studied, at baseline and after treatment with interferon alpha (21 cases) or pegylated interferon (6 cases) plus ribavirin. A group of 15 healthy sex‐ and age‐matched individuals was selected as control. Serum concentrations of sFas, IL‐10 and IL‐18 were determined by ELISA in sandwich. The relationship of these molecules to necro‐inflammatory and fibrotic activity was evaluated. Evolution of the serum concentrations of these molecules was analysed after treatment. Significantly increased serum concentrations of sFas were detected in patients with chronic hepatitis, compared with controls. Levels of this molecule were significantly correlated with necroinflammatory activity. Likewise, concentrations of IL‐10 were significantly increased in the group of patients, compared with controls. Treatment with interferon and ribavirin induced a significant decrease of IL‐18 concentration independently of the viral response. In contrast, levels of sFas decreased only in those patients with sustained response to therapy. Finally, baseline levels of IL‐10 were significantly increased in patients without response to treatment, compared with those with sustained response, but the concentration did not change with the treatment. Increased serum levels of IL‐10 are a negative prognostic marker of response to hepatitis C treatment. A significant decrease of apoptotic rate, as determined by sFas, can be expected in patients with a response to therapy.
Pegylated interferon and ribavirin treatment for hepatitis C in haemodialysis patients
Authors: Bruchfeld A. , Lindahl K. , Reichard O. , Carlsson T. , Schvarcz R.
Published: October 2005

Summary.  Standard therapy for chronic hepatitis C (HCV) is pegylated interferon in combination with ribavirin. There is limited experience with either drug in dialysis [end stage renal disease (ESRD)]. Six haemodialysis patients, four with HCV genotype 1, one with genotype 4 and one genotype 2 were treated with pegylated interferon‐alfa‐2b (n = 4) and pegylated interferon‐alfa‐2a (n = 2) for 24–48 weeks according to genotype with a dose of 50 or 135 μg/week respectively. All patients were given reduced ribavirin doses, initially 200–400 mg/day. Ribavirin trough plasma concentrations were measured with a HPLC method previously developed for earlier treatment studies, aiming at a target concentration of 10–15 μmol/L. Interferon related side‐effects were common, in one patient peg‐alfa‐2b was permanently reduced to 50 μg every 9–10 days with improvement in tolerance. Average ribavirin dose was 170–300 mg/day. Ribavirin‐induced anaemia was treated with high doses of erythropoietin and low doses of iron. Blood‐transfusions were not needed. All patients became HCV‐RNA‐PCR negative during treatment which was completed or nearly completed in four patients. One patient terminated therapy prematurely due to pronounced interferon related side‐effects and another died of myocardial infarction probably not related to therapy. Three patients have remained HCV‐RNA negative with extended follow‐up, two of whom have had a successful kidney transplant. Pegylated interferons are likely to become a valuable addition for HCV therapy in ESRD and are possible to combine with ribavirin. However the pharmacokinetics and tolerability of both peg‐alfa‐2a and 2b need to be studied more closely in prospective studies before definite dosing recommendations can be made.
Comparison of hepatitis C treatment patterns in patients with and without psychiatric and/or substance use disorders†
Authors: Chainuvati S. , Khalid S. K. , Kancir S. , Shea M. , Edwards J. , Sernyak M. , Wongcharatrawee S. , Garcia‐Tsao G.
Published: October 2005

Summary.  Hepatitis C virus (HCV) infection is more frequent in veterans than in nonveterans. Up to 85% of HCV‐infected veterans have psychiatric and/or substance use (SU) co‐morbidities which, prior to the 2002 NIH Consensus Conference, were considered relative contraindications to antiviral therapy, assuming a poor adherence. With the objective of evaluating the validity of this assumption, we compared eligibility, completion and response to antiviral therapy in HCV‐infected veterans with and without these comorbidities. Veterans who were anti‐HCV‐positive and had been seen at least once in the liver clinic (between October 1999 and June 2002) were identified through the CT‐VAHCS database. Records were reviewed for patient demographics and status of liver disease, assessment of treatment eligibility, type of therapy, completion of therapy and virological response. Patients with active mental illness (MI) or SU were compared with those without these comorbidities (controls). Of 697 anti‐HCV‐positive‐patients, 647 HCV‐RNA‐positive patients were included, 294 with MI/SA and 353 controls. Patient demographics, viral and liver disease characteristics were comparable between groups. Patients with MI/SA were considered ineligible for therapy more frequently (53%vs 39%, P < 0.001) and were treated less frequently (21%vs 28%, P = 0.03) than controls. However, completion of therapy (72%vs 59%) and sustained virological response (SVR) (20%vs 25%) did not differ significantly between groups. HCV‐infected veterans with MI/SA are being offered therapy and treated less often than those without such co‐morbidities, however therapy completion and SVR rates are similar, challenging the perception that adherence is poorer in this patient population.
Sustained virological response rates and durability of the response to interferon‐based therapies in hepatitis C patients treated in the clinical setting
Authors: Desmond C. P. , Roberts S. K. , Dudley F. , Mitchell J. , Day C. , Nguyen S. , Pianko S.
Published: October 2005

Summary.  International controlled trials have demonstrated increasing sustained virological response (SVR) rates to interferon‐based therapies in hepatitis‐C‐treated patients. Response rates of 6–20% in the era of interferon monotherapy are compared with 42–82% with pegylated interferon plus ribavirin. The virological durability of the SVR is unknown and the optimal follow‐up for these patients is unclear. The aim of our study was to determine SVR rates and the durability of the response to interferon‐based therapies in the clinical setting. From our database of 1540 hepatitis C patients, 344 treatment courses of at least 12 weeks duration were identified, including interferon monotherapy (175 patients), interferon plus ribavirin (96 patients) and peginterferon plus ribavirin (73 patients). Interferon monotherapy was associated with an SVR rate of 5% in 103 genotype 1 patients and 25% in 72 genotype 2/3 patients. Response rates were higher (P < 0.001) with interferon plus ribavirin—41% in 34 genotype 1 patients and 73% in 62 genotype 2/3 patients—and with peginterferon plus ribavirin—47% in 47 genotype 1 patients and 79% in 26 genotype 2/3 patients. Of 147 patients with an SVR, 146 (>99%) remained hepatitis C virus PCR negative during a mean 2.3 years (range 0.3–10.3) of follow‐up. In conclusion, with advances in therapies, we are achieving higher response rates in hepatitis C patients treated in the clinical setting. We can now expect an SVR in over half of the treated patients. Importantly, the response is durable and medium and long‐term follow‐up of these patients are of low yield and largely unnecessary.
Treatment of lymph node tuberculosis – a randomized clinical trial of two 6‐month regimens
Authors: Jawahar M. S. , Rajaram K. , Sivasubramanian S. , Paramasivan C. N. , Chandrasekar K. , Kamaludeen M. N. , Thirithuvathas A. J. , Ananthalakshmi V. , Prabhakar R.
Published: October 2005

Objective  The currently recommended treatment for lymph node tuberculosis is 6 months of rifampicin and isoniazid plus pyrazinamide for the first 2 months, given either daily or thrice weekly. The objective of this study was to assess the efficacy of a 6‐month twice‐weekly regimen and a daily two‐drug regimen.

Methods  Patients with biopsy confirmed superficial lymph node tuberculosis were randomly allocated to receive either a daily self‐administered 6‐month regimen of rifampicin and isoniazid, or a twice‐weekly, directly observed, 6‐month regimen of rifampicin and isoniazid plus pyrazinamide for the first 2 months, in Madurai, South India, Patients were followed up for 36 months after completing treatment.

Results  Of 277 enrolled patients, data was available for analysis in 268. At the end of treatment, 116 of 134 [87%; 95% confidence interval (CI) 81–93%] patients in each treatment group had a favourable clinical response; 14 (11%; 95% CI 6–16%) and 17 (13%; 95% CI 7–19%) patients had a doubtful response, and 4 (3%; 95% CI 0–6%) and 1 (1%; 95% CI 0–2%) patients had an unfavourable response among those treated with the daily and twice‐weekly regimen, respectively. During 36 months after completion of treatment, five patients [2 (2%; 95% CI 1–3%) and 3 (2%; 95% CI 1–3%) patients treated with the daily and twice‐weekly regimen, respectively] had relapse of lymph node tuberculosis, of 260 assessed. Adverse reactions probably attributable to the treatment regimens occurred in 1% of the patients treated daily and in 11% of those treated twice‐weekly (P < 0.001). At the end of 36 months after treatment, 126 of 134 (94%; 95% CI 90–98%) and 129 of 134 (96%; 95% CI 94–98%) of the patients treated with the daily and twice‐weekly regimen, respectively, had a successful outcome.

Conclusion  Both the self‐administered daily regimen and the fully observed twice‐weekly regimen were highly efficacious for treating patients with lymph node tuberculosis and may be considered as alternative options to the recommended regimens.
Repellent properties of celery, Apium graveolens L., compared with commercial repellents, against mosquitoes under laboratory and field conditions
Authors: Tuetun Benjawan , Choochote Wej , Kanjanapothi Duangta , Rattanachanpichai Eumporn , Chaithong Udom , Chaiwong Prasong , Jitpakdi Atchariya , Tippawangkosol Pongsri , Riyong Duangrat , Pitasawat Benjawan
Published: October 2005

In our search for new bioactive products against mosquito vectors, we reported the slightly larvicidal and adulticidal potency, but remarkable repellency of Apium graveolens both in laboratory and field conditions. Repellency of the ethanolic preparation of hexane‐extracted A. graveolens was, therefore, investigated and compared with those of 15 commercial mosquito repellents including the most widely used, DEET. Hexane‐extracted A. graveolens showed a significant degree of repellency in a dose‐dependent manner with vanillin added. Ethanolic A. graveolens formulations (10–25% with and without vanillin) provided 2–5 h protection against female Aedes aegypti. Repellency that derived from the most effective repellent, 25% of hexane‐extracted A. graveolens with the addition of 5% vanillin, was comparable to the value obtained from 25% of DEET with 5% vanillin added. Moreover, commercial repellents, except formulations of DEET, showed lower repellency than that of A. graveolens extract. When applied on human skin under field conditions, the hexane‐extracted A. graveolens plus 5% vanillin showed a strong repellent action against a wide range of mosquito species belonging to various genera. It had a protective effect against Aedes gardnerii, Aedes lineatopennis, Anopheles barbirostris, Armigeres subalbatus, Culex tritaeniorhynchus, Culex gelidus, Culex vishnui group and Mansonia uniformis. The hexane‐extracted A. graveolens did not cause a burning sensation or dermal irritation when applied to human skin. No adverse effects were observed on the skin or other parts of the human volunteers’ body during 6 months of the study period or in the following 3 months, after which time observations ceased. Therefore, A. graveolens can be a potential candidate for use in the development of commercial repellents that may be an alternative to conventional synthetic chemicals, particularly in community vector control applications.
Integrating insecticide‐treated bednets into a measles vaccination campaign achieves high, rapid and equitable coverage with direct and voucher‐based methods
Authors: Grabowsky Mark , Farrell Nick , Hawley William , Chimumbwa John , Hoyer Stefan , Wolkon Adam , Selanikio Joel
Published: October 2005

Population coverage of insecticide‐treated nets (ITNs) in Africa falls well below the Abuja target of 60% while coverage levels achieved during vaccination campaigns in the same populations typically exceed 90%. Household (HH) cost of ITNs is an important barrier to their uptake. We investigated the coverage, equity and cost of linking distribution of free ITNs to a measles vaccination campaign. During a national measles vaccination campaign in Zambia, children in four rural districts were given a free ITN when they received their measles vaccination. In one urban district, children were given a voucher, which could be redeemed for a net at a commercial distribution site. About 1700 HHs were asked whether they received vaccination and an ITN during a measles campaign, as well as questions on assets (e.g. type roofing material or bicycle ownership) to assess HH wealth. Net ownership was calculated for children in each wealth quintile. In the rural areas, ITN coverage among children rose from 16.7% to 81.1% and the equity ratio from 0.32 to 0.88 and in the urban area from 50.7% to 76.2% (equity ratio: 0.66–1.19). The operational cost per ITN delivered was $0.35 in the rural area with direct distribution and $1.89 in the urban areas with voucher distribution. Mass distribution of ITNs through vaccination campaigns achieves rapid, high and equitable coverage at low cost.
Open randomized study of artesunate‐amodiaquine vs. chloroquine‐pyrimethamine‐sulfadoxine for the treatment of uncomplicated Plasmodium falciparum malaria in Nigerian children
Authors: Sowunmi A. , Fehintola F. A. , Adedeji A. A. , Gbotosho G. O. , Tambo E. , Fateye B. A. , Happi T. C. , Oduola A. M. J.
Published: October 2005

Background  Artemisinin‐based combination antimalarials are currently considered effective alternatives for the treatment of malaria in Africa, but there are few studies of such combinations in Nigerian children. We assessed the safety, treatment efficacy and effects on gametocyte carriage of the combination of artesunate plus amodiaquine and chloroquine plus pyrimethamine‐sulfadoxine in children.

Methods  We evaluated 153 children who were aged 12 years or younger who had uncomplicated Plasmodium falciparum malaria. Patients were randomly assigned a combination of artesunate (4 mg/kg of body weight daily for 3 days) plus amodiaquine (30 mg/kg over 3 days), or chloroquine (25 mg/kg over 3 days) plus pyrimethamine‐sulfadoxine (25 mg/kg of the sulfadoxine component at presentation). The primary endpoints were the proportions of children with adequate clinical and parasitological response, late parasitological failure, late clinical failure and early treatment failure. The parasitological cure rates on days 14–28 were also used as the primary endpoints.

Results  Both regimens were well tolerated; no child was withdrawn because of drug intolerance. All children treated with artesunate plus amodiaquine had adequate clinical and parasitological response (ACPR), while all but five children treated with chloroquine plus pyrimethamine‐sulfadoxine had similar response. Fever clearance times were similar in the two treatment groups. However, the proportion of patients whose parasitaemia cleared by day 2 was significantly higher (100 vs. 50%, P = 0.00001) and parasite clearance was significantly faster (1.7 ± 0.4 vs. 2.5 ± 0.8 days, P = 0.0001) in children treated with artesunate plus amodiaquine. The cure rates on days 21 (100%vs. 94%, P = 0.03) and 28 (100%vs. 90%, P = 0.003) were also significantly higher in children treated with artesunate plus amodiaquine than in those treated with chloroquine plus pyrimethamine‐sulfadoxine. Overall, a significantly higher proportion of children treated with chloroquine plus pyrimethamine‐sulfadoxine carried gametocytes at least once during follow‐up compared with those treated with artesunate plus amodiaquine [5 of 50 (10%) vs. 1 of 103 (0.97%), P = 0.01].

Conclusion  The combination of artesunate plus amodiaquine is therapeutically superior to a combination of chloroquine plus pyrimethamine‐sulfadoxine, and significantly reduced gametocyte carriage following treatment.
Mutations in Plasmodium falciparum dihydrofolate reductase and dihydropteroate synthase genes in Senegal
Authors: Ndiaye D. , Daily J. P. , Sarr O. , Ndir O. , Gaye O. , Mboup S. , Wirth D. F.
Published: October 2005

Senegal recently (2004) switched to sulfadoxine‐pyrimethamine (SP) with amodiaquine as first line therapy for malaria in response to increasing chloroquine resistance. In anticipation of emerging resistance to SP as a result of this change in drug pressure, we set out to define the baseline prevalence of SP‐associated mutations in the dhfr and dhps genes in Plasmodium falciparum using geographically diverse and longitudinally collected samples. A total of 153 blood samples were analysed from patients (5 years or older) with mild malaria after informed consent was obtained. Longitudinal samples were collected between 2000 and 2003 in Pikine, a suburb of Dakar. Geographically diverse site sampling was carried out in 2003. The mutation prevalence in DHFR codons 51, 59 and 108 is 65%, 61% and 78% in Pikine, 2003. The overall prevalence of the triple mutation that is associated with high‐level pyrimethamine resistance is 61%. The mutation prevalence rate in DHPS codons 436 and 437 is 21% and 40%, respectively. There is significant geographic variation in genotypic resistance, as samples from Pikine in 2003 had higher mutation prevalence in the pfdhfr and pfdhps genes compared to samples from Tambacounda (P < 0.015). In summary, this study demonstrates a high background prevalence of SP resistance mutations already present in P. falciparum in Senegal.