bluuebunny/crossref_metadata_2025 huggingface dataset:
Metabolic flux analysis in plants: coping with complexity
Authors: ALLEN DOUG K. , LIBOUREL IGOR G. L. , SHACHAR‐HILL YAIR
Published: April 2009

Theory and experience in metabolic engineering both show that metabolism operates at the network level. In plants, this complexity is compounded by a high degree of compartmentation and the synthesis of a very wide array of secondary metabolic products. A further challenge to understanding and predicting plant metabolic function is posed by our ignorance about the structure of metabolic networks even in well‐studied systems. Metabolic flux analysis (MFA) provides tools to measure and model the functioning of metabolism, and is making significant contributions to coping with their complexity.

This review gives an overview of different MFA approaches, the measurements required to implement them and the information they yield. The application of MFA methods to plant systems is then illustrated by several examples from the recent literature. Next, the challenges that plant metabolism poses for MFA are discussed together with ways that these can be addressed. Lastly, new developments in MFA are described that can be expected to improve the range and reliability of plant MFA in the coming years.
Dynamics of vacuoles and actin filaments in guard cells and their roles in stomatal movement
Authors: GAO XIN‐QI , WANG XIU‐LING , REN FEI , CHEN JIA , WANG XUE‐CHEN
Published: April 2009

Vacuoles and actin filaments are important cytoarchitectures involved in guard cell function. The changes in the morphology and number of vacuoles and the regulation of ion channel activity in tonoplast of guard cells are essential for stomatal movement. A number of studies have investigated the regulation of ion channels in animal and plant cells; however, little is known about the regulating mechanism for vacuolar dynamics in stomatal movement. Actin filaments of guard cells are remodelling with the changes in the stomatal aperture; however, the dynamic functions of actin filaments in stomatal movement remain elusive. In this paper, we summarize the recent developments in the understanding of the dynamics of actin filaments and vacuoles of guard cells during stomatal movement. All relevant studies suggest that actin filaments might be involved in stomatal movement by regulating vacuolar dynamics and the ion channels in tonoplast. The future study could be focused on the linker protein mediating the interaction between actin filaments and tonoplast, which will provide insights into the interactive function of actin and vacuole in stomatal movement regulation.
GhDREB1 enhances abiotic stress tolerance, delays GA‐mediated development and represses cytokinin signalling in transgenic Arabidopsis
Authors: HUANG JIN‐GUANG , YANG MEI , LIU PEI , YANG GUO‐DONG , WU CHANG‐AI , ZHENG CHENG‐CHAO
Published: April 2009

Plants vary significantly in their ability to tolerate low temperatures. The CBF/DREB1 cold response pathway has been identified in many plant species and plays a pivotal role in low temperature tolerance. Here, we show that GhDREB1 is a functional homologue and elevates the freezing, salt and osmotic stress tolerance of transgenic Arabidopsis. The constitutive expression of GhDREB1 in Arabidopsis caused dwarfism and late flowering phenotypes, which could be rescued by exogenous application of GA3. Endogenous bioactive GA contents were significantly lower in GhDREB1 overexpressing Arabidopsis than in wild‐type plants. RT‐PCR analyses revealed that the transcript levels of the GA synthase genes were higher in transgenics than in wild‐type plants, whereas the GA deactivating genes were lower. Flowering related genes in different regulatory pathways were also affected by GhDREB1, which may account for the flowering delay phenotype. Moreover, the GhDREB1 overexpressing Arabidopsis exhibited decreased sensitivity to cytokinin (CK) which is associated with repression of expression of type‐B and type‐A ARRs, two key components in the CK‐signalling pathway.
High‐frequency cyclicity (Milankovitch and millennial‐scale) in slope‐apron carbonates: Zechstein (Upper Permian), North‐east England
Authors: MAWSON MIKE , TUCKER MAURICE
Published: April 2009

The Upper Permian (Zechstein) slope carbonates in the Roker Formation (Zechstein 2nd‐cycle Carbonate) in North‐east England consist of turbidites interbedded with laminated lime‐mudstone. Studies of turbidite bed thickness and relative proportion of turbidites (percentage turbidites in 20 cm of section) reveal well‐developed cyclicities consisting of thinning‐upward and thickening‐upward packages of turbidite beds. These packages are on four scales, from less than a metre, up to 50 m in thickness. Assuming that the laminae of the hemipelagic background sediment are annual allows the durations of the cycles to be estimated. In addition, counting the number and thickness of turbidite beds in 20 cm of laminated lime‐mudstone, which is approximately equivalent to 1000 years (each lamina is 200 μm), gives the frequencies of the turbidite beds, the average thicknesses and the overall sedimentation rates through the succession for 1000 year time‐slots. Figures obtained are comparable with modern rates of deposition on carbonate slopes. The cyclicity present in the Roker Formation can be shown to include: Milankovitch‐band ca 100 kyr short‐eccentricity, ca 20 kyr precession and ca 10 kyr semi‐precession cycles and sub‐Milankovitch millennial‐scale cycles (0·7 to 4·3 kyr). Eccentricity and precession‐scale cycles are related to ‘highstand‐shedding’ and relative sea‐level change caused by Milankovitch‐band orbital forcing controlling carbonate productivity. The millennial‐scale cycles, which are quasi‐periodic, probably are produced by environmental changes controlled by solar forcing, i.e. variations in solar irradiance, or volcanic activity. Most probable here are fluctuations in carbonate productivity related to aridity–humidity and/or temperature changes. Precession and millennial‐scale cycles are defined most strongly in early transgressive and highstand parts of the larger‐scale short‐eccentricity cycles. The duration of the Roker Formation as a whole can be estimated from the thickness of the laminated lithotype as ca 0·3 Myr.
A quantitative analysis of temperature‐dependent dormancy changes inPolygonum aviculareseeds
Authors: BATLLA D , GRUNDY A , DENT K C , CLAY H A , FINCH‐SAVAGE W E
Published: April 2009

Mathematical models that predict emergence of weed seedbanks could be useful tools for determining the most suitable time for weed seedling control and, consequently, should result in a higher efficacy of applied control methods. To achieve this goal in dormant weed species, functional relationships should be established between environmental factors regulating dormancy and dormancy changes of seed populations. In the present work, we used a simple model and an optimisation procedure to quantify the effect of temperature onPolygonum aviculareseed dormancy release and induction, based on germination data. Dormancy release rate was inversely related to temperature, showing a decreasing logistic trend that results in no dormancy release for seeds exposed to 20 and 25°C. In contrast, dormancy induction rates in absolute values were positively associated with temperature, showing a logistic trend in which dormancy induction was almost zero at low temperatures and maximal at 25°C. Derived model parameters were used to simulate dormancy changes ofP. aviculareseeds stored under controlled and field conditions. These results suggest that similar model structures could be used to quantify temperature effects on seed dormancy status of other weed species and to develop predictive models of weed emergence.
The efficiency of a vacuum device for estimating soil‐surface seed abundance on lowland farms
Authors: EVANS D M , MCLEOD J J , PASCOE L , MEMMOTT J
Published: April 2009

Accurate sampling methods for estimating soil‐surface seed abundance in agro‐ecosystems are vital for understanding arable plant population dynamics, as well as quantifying seed availability as a food resource for farmland birds, mammals and arthropods. We tested the sampling efficiency of a portable, petroleum spirit powered leaf blower‐vacuum in recovering four different seed size classes on the soil‐surface of seven lowland farm habitats. Vacuum sampler efficiency was dependent on seed size, habitat type and exposure of seeds to environmental conditions. Larger seeds were more readily recovered and overall seed recovery was consistently higher on habitats with low vegetation cover, such as newly‐sown cereal fields. We discuss the limitations of the device, but commend its use with correction factors to estimate soil‐surface seed abundance in agricultural habitats. This study highlights the potential biases when estimating soil‐surface seed abundance, the importance of correcting such biases and the need for better sampling techniques for weed science.
Breastfeeding and atopic eczema in Japanese infants: The Osaka Maternal and Child Health Study
Authors: Miyake Yoshihiro , Tanaka Keiko , Sasaki Satoshi , Kiyohara Chikako , Ohya Yukihiro , Fukushima Wakaba , Yokoyama Tetsuji , Hirota Yoshio ,   for the Osaka Maternal and Child Health Study Group
Published: April 2009

Epidemiological studies associated with breastfeeding have provided conflicting results about whether it is preventive or a risk factor for atopic eczema in children. The current prospective study investigated the relationship between breastfeeding and the risk of atopic eczema in Japan. A birth cohort of 763 infants was followed. The first survey during pregnancy and the second survey between 2 and 9 months postpartum collected information on potential confounding factors and atopic eczema status. Data on breastfeeding and symptoms of atopic eczema were obtained from questionnaires in the third survey from 16 to 24 months postpartum. The following variables were a priori selected as potential confounders: maternal age, maternal and paternal history of asthma, atopic eczema, and allergic rhinitis, indoor domestic pets (cats, dogs, birds, or hamsters), family income, maternal and paternal education, maternal smoking during pregnancy, baby’s sex, baby’s birth weight, baby’s older siblings, household smoking in the same room as the infant, and time of delivery before the third survey. In the third survey, 142 infants (18.6%) were revealed to have developed atopic eczema based on criteria of the International Study of Asthma and Allergies in Childhood. In an overall analysis, neither exclusive nor partial breastfeeding was significantly related to the risk of atopic eczema. After excluding 64 infants identified with suspected atopic eczema in the second survey, both exclusive breastfeeding for 4 months or more and partial breastfeeding for 6 months or more were independently associated with an increased risk of atopic eczema only among infants with no parental history of allergic disorders [multivariate odds ratios were 2.41 (95% confidence interval, 1.10–5.55) and 3.39 (95% confidence interval, 1.20–12.36), respectively]. The authors found that, overall, neither exclusive nor partial breastfeeding had a strong impact on the risk of atopic eczema. However, a parental allergic history may affect the risk.
Association of lower eosinophil‐related T helper 2 (Th2) cytokines with coronary artery lesions in Kawasaki disease
Authors: Kuo Ho‐Chang , Wang Chih‐Lu , Liang Chi‐ Di , Yu Hong‐Ren , Huang Chien‐Fu , Wang Lin , Hwang Kao‐Pin , Yang Kuender D.
Published: April 2009

Kawasaki disease (KD) is a systemic febrile vasculitis particular coronary artery involvement. Eosinophilia has been found in our and other studies in KD. This study further investigates whether eosinophil‐related T helper 2 (Th2) cytokines or the activation marker (eosinophil cationic protein – ECP) is involved in KD with coronary artery lesions (CAL). A total of 95 KD patients were enrolled for this study. Plasma samples were subjected to the measurement of interleukin (IL)‐4, IL‐5, and eotaxin by Luminex‐Bedalyte multiplex beadmates system and to the measurement of ECP by fluoroimmunoassay. Patients with KD had higher eosinophils than controls. Eosinophil‐related mediators: IL‐4, IL‐5, eotaxin, and ECP levels were also higher in KD patients than controls before intravenous immunoglobulin (IVIG) treatment. After IVIG treatment, ECP decreased but IL‐4, IL‐5, and eotaxin increased significantly. The higher the IL‐5 and eosinophil levels after IVIG treatment, the lower rate of CAL was found. Changes of eosinophils after IVIG treatment were positively correlated to changes of IL‐5 levels but not ECP levels. An increase of eosinophils and IL‐5, but not ECP levels after IVIG treatment, was inversely correlated with CAL formation in KD.
Association of cord blood cytokine levels with wheezy infants in the first year of life
Authors: Tadaki Hiromi , Arakawa Hirokazu , Sugiyama Mikio , Ozawa Kiyoshi , Mizuno Takahisa , Mochizuki Hiroyuki , Tokuyama Kenichi , Morikawa Akihiro
Published: April 2009

As antenatal environment may influence the development of atopy‐predisposing immune response, cord blood cytokine productions may be an important predictor for wheezing. We investigated cord blood cytokines in a prospective birth cohort, intensively monitored for wheezy infant outcome at 1 yr. Cord blood serum samples from 269 children were assayed for interleukin (IL)‐1β, ‐2, ‐4 to ‐8, ‐10, ‐12 (p70), ‐13, and ‐17, interferon‐γ, tumor necrosis factor‐α, granulocyte‐macrophage colony‐stimulating factor, granulocyte colony‐stimulating factor (G‐CSF), monocyte chemotactic protein‐1, and macrophage inflammatory protein‐1β. Associations between family histories, antenatal and perinatal factors, cord blood cytokine concentrations, and wheezy infant outcomes (wheezing more than two times) were analyzed. In cord blood sera from 269 children, there were associations between high levels of IL‐6, ‐8 and G‐CSF concentrations, and cesarean section. Data at 1 yr were obtained from 213 infants, including 33 wheezy infants. Risk of wheezing was related to gestational age, birth weight, cesarean section, and maternal eczema, but not to bacterial/viral infection during pregnancy, maternal asthma, maternal smoking, or paternal history. High level of cord blood IL‐8 concentration had a significant association with wheezy infant outcomes at 1 yr (p = 0.025). By using multivariate logistic regression analysis, birth weight [odds ratio(OR) = 0.998, 95% confidence interval (CI) = 0.997–1.000] and maternal eczema (OR = 5.356, 95% CI = 1.340–21.41), but no other factors, were significant predictors of wheezy infants. Birth weight, gestational age, and maternal history were important risk factors for wheezing in the first year of life. Several cord blood cytokine productions were influenced by cesarean section, and IL‐8 may be a predictor for recurrent wheezing at 1 yr.
Increased rate and greater severity of allergic reactions to insect sting among schoolchildren with atopic diseases
Authors: Graif Yael , Romano‐Zelekha Orly , Livne Irit , Green Manfred S. , Shohat Tamy
Published: April 2009

The question of whether atopic diseases are a risk factor for allergic reactions to insect sting is still unresolved. The aim of this study was to evaluate the association between atopic diseases (asthma, allergic rhinitis, atopic eczema) and allergic reactions to insect stings among schoolchildren in Israel. A self‐report questionnaire of the International Study of Asthma and Allergies in Childhood was administered to a national sample of 13–14‐yr‐old schoolchildren. Questions regarding reactions to insect stings were added. A total of 10,021 questionnaires were available for analysis. Among the children who reported insect stings (56.3%), the prevalence of current asthma was 6.0%, of allergic rhinitis, 10.5%, and of atopic eczema, 8.7%, with no significant differences from the whole study population. Among children with any of the atopic diseases, 36.9% reported an allergic reaction to insect sting compared to 24.8% of the non‐atopic children (p < 0.0001). On multivariate analysis, asthma, allergic rhinitis, and atopic eczema were found to be significant risk factors for allergic reactions of any severity. Children in the atopic group had a significantly higher rate of severe allergic reactions than the non‐atopic children, and relatively higher rates of milder ones (p < 0.0001). Asthmatic patients with severe allergic reactions had more parameters of severe asthma than asthmatic patients with mild or no reactions. In conclusions, allergic diseases are associated with a higher rate and greater severity of allergic reactions to insect sting in children. The severity of the allergic reaction is related to the severity of the asthma symptoms.
Maturation of cytokine‐producing capacity from birth to 1 yr of age
Authors: Lappalainen Mikko , Roponen Marjut , Pekkanen Juha , Huttunen Kati , Hirvonen Maija‐Riitta
Published: April 2009

Little is known about the immunologic maturation in the early stages of life. The aim of this study was to investigate maturation of immune system from birth to 1 yr of age and to compare immune functions between mothers and their children. Also the effect of atopy to the immune responses of children was examined. Cord blood samples (n = 228) and peripheral blood samples of children (n = 200) and their mothers (n = 208) 1 yr after birth were collected. Whole blood samples were stimulated for 24 and 48 h with Staphylococcal enterotoxin B (SEB), lipopolysaccharide (LPS) and the combination of phorbol ester and ionomycin (P/I). Production of TNF‐α, IFN‐γ, IL‐5, IL‐8 and IL‐10 was determined using ELISA. Significant mother‐to‐child correlation was detected in cytokine‐producing capacity at the age of 1 yr. TNF‐α (P/I, SEB and LPS stimulation), IFN‐γ (P/I and SEB), IL‐5 (P/I and SEB) and IL‐10 (P/I, SEB and LPS) producing capacity increased from birth to 1 yr of age. In general, stimulated cytokine responses were higher in mothers’ than in children’s blood samples, except in the case of P/I and LPS‐stimulated IL‐8, which were highest at birth. Maternal inhalation atopy was associated with increased cord blood IL‐5 (24 and 48 h) and IL‐10 (48 h) production following P/I stimulation. Also children of food atopic mothers expressed elevated cord blood IL‐10 (48 h, P/I) responses and decreased IFN‐γ/IL‐5 ratio (24 h, P/I). In addition, the production of IFN‐γ (24 and 48 h, P/I) and the IFN‐γ/IL‐5 ratio (24 h and 48 h, P/I) at the age of 1 yr was lower among children with food atopic mothers. In conclusion, our results suggest that both adaptive and innate immune responses increase from birth to 1 yr of age, but are still weak in comparison to adult responses. Cytokine responses of children begin to correlate with those of their mothers during the first year of life. Although only few associations were observed between atopy and cytokine‐producing capacity, our results suggest that children of atopic mothers express Th2‐polarized cytokine pattern.
Time trends of co‐existing atopic conditions in Singapore school children: prevalence and related factors
Authors: Wang Xiao Shan , Shek Lynette P. , Ma Stefan , Soh Shu E , Lee Bee Wah , Goh Daniel Y. T.
Published: April 2009

Wang XS, Shek LP, Ma S, Soh SE, Lee BW, Goh DYT. Time trends of co‐existing atopic conditions in Singapore school children: prevalence and related factors. Pediatr Allergy Immunol 2010: 21: e137–e141. © 2009 The Authors Journal compilation © 2009 Blackwell Munksgaard

Atopic diseases are known to co‐exist and are interrelated. Their co‐existence in a patient increases disease burden and morbidity. We sought to study the prevalence of co‐existing atopic conditions over time and its associated risk factors among Singapore children. Data was collected from two respective The International Study of Asthma and Allergies in Childhood surveys conducted in 1994 and 2001. Data on asthma, rhinitis and eczema were obtained from the responses to questions about wheezing, exercise‐induced wheezing, nocturnal cough, rhinitis symptoms, rash, and diagnoses of asthma, rhinitis and eczema in the last 12 months. Data on demographic and known risk factors were also obtained. The prevalence of children who have had more than one atopic disorder increased significantly from 6.0% in 1994 to 10.2% in 2001 (p < 0.001). Parental history of atopic disease, maternal educational level, male gender and smoke exposure were significantly associated with co‐existent atopic symptoms in the child. Among Singapore schoolchildren, there was a significant prevalence of co‐existent atopic disease and this prevalence was rising. The influences of both genetic and environmental factors were likely to have contributed to these observations.
A novel splicing variant encoding putative catalytic α subunit of maize protein kinase CK2
Authors: Łebska Maja , Szczegielniak Jadwiga , Dobrowolska Grażyna , Cozza Giorgio , Moro Stefano , Muszyńska Grażyna
Published: April 2009

A cDNA highly homologous to the known catalytic α subunit of protein kinase CK2 was cloned from maize (Zea mays). It was designated ZmCK2α‐4 (accession no. AAF76187). Sequence analysis shows that ZmCK2α‐4 and the previously identified ZmCK2α‐1 (accession no. X61387) are transcribed from the same gene, ZmPKCK2AL (accession no. Y11649), but at different levels in various maize organs and at different stages of development. The cDNA encoding ZmCK2α‐4 has three potential translation initiation sites. The three putative variants of ZmCK2α‐4 were expressed in Escherichia coli as GST‐fusion proteins and purified from bacterial extracts. In contrast to the previously characterized ZmCK2αs, the obtained GST:ZmCK2α‐4 proteins were catalytically inactive as monomers or in the presence of equimolar amounts of the human CK2β. However, GST:ZmCK2α‐4 did phosphorylate casein in the presence of a large excess of the β subunit. The activity of ZmCK2α‐4 toward casein could also be stimulated by increasing ATP concentration. Modeling studies have shown that there is no interaction between the N‐terminal segment of ZmCK2α‐4 and the activation loop responsible for constitutive catalytic activity of CK2α. Preliminary results suggest that ZmCK2α‐4 may function as a negative regulator of other CK2s, and at certain circumstances as a holoenzyme which catalytic activity is stimulated by specific regulatory subunit(s).
Alternative oxidase: a target and regulator of stress responses
Authors: Van Aken Olivier , Giraud Estelle , Clifton Rachel , Whelan James
Published: April 2009

The alternative oxidase (AOX) is found in all plants examined to date, fungi and lower invertebrates. We propose that AOX is not only part of the stress response in plants, but it also plays a central role in defining the stress response. Three lines of evidence support this proposal: (1) The absence of AOX leads to an alteration of stress defences in normal and stress conditions, (2) the expression of AOX is triggered by a variety of signals indicating that it is a common response and (3) AOX acts as a buffer that determines the threshold for the induction of programmed cell death. Therefore, AOX is not only one of many components involved in the defence response, its activity or lack of activity leads to a radical alteration of the defence equilibrium at a cellular level and thus it plays a central role in programming the stress response. This programming role of AOX can be achieved directly by its ability to suppress the induction of reactive oxygen species (ROS) and indirectly by causing changes in the energy status of cells owing to the non‐phosphorylating nature of the alternative respiratory pathway. The latter is likely achieved in combination with a variety of alternative NAD(P)H dehydrogenases, that are co‐regulated with AOX. Additionally, we explore the possible function of AOX as a component of the stress response beyond the plant frontier.
Distribution and expression of porcine endogenous retroviruses in multi‐transgenic pigs generated for xenotransplantation
Authors: Dieckhoff Britta , Kessler Barbara , Jobst Danny , Kues Wilfried , Petersen Björn , Pfeifer Alexander , Kurth Reinhard , Niemann Heiner , Wolf Eckhard , Denner Joachim
Published: April 2009

Background:  Multi‐transgenic pigs produced for use in xenotransplantation have to be screened for the presence and expression of porcine endogenous retroviruses (PERV) to select animals with low PERV load. The production of transgenic pigs may also be associated with the integration of the transgene adjacent to or into the locus of a PERV provirus, potentially leading to an enhanced virus expression.

Methods:  Non‐transgenic animals, single‐transgenic, and multi‐transgenic pigs were screened for the presence of PERV‐A, ‐B, and ‐C and recombinant PERV‐A/C using polymerase chain reaction (PCR). PERV expression was determined by real time reverse transcriptase‐PCR. An assay based on the activation of PERV in peripheral blood mononuclear cells by mitogens was used to discriminate between low and high PERV producer animals.

Results:  All animals carried PERV‐A and ‐B. A total of 176 from 181 (97.2%) animals carried PERV‐C in the germ line and 18 from 64 animals carried PERV‐A/C in the genome of lymphoid cells but not in the germ line. The expression of PERV was very low in all animals and not different between transgenic pigs and non‐transgenic animals. PERV expression differed between various pig lines. The highest expression was found in mini‐pigs and crossing other pig lines with mini‐pigs resulted in increased PERV expression in the progeny. However, expression of viral proteins and particle release were not observed in all transgenic animals.

Conclusions:  No evidence for elevated PERV expression in (multi‐) transgenic pigs was observed. Differences in PERV expression correlated with the genetic background of the animals, not with the specific transgene. Mini‐pigs consistently had the highest level of PERV expression and animals with a mini‐pig background had a higher level of expression compared with animals without mini‐pig background.
In vitro and in vivo effects of the carbon monoxide‐releasing molecule, CORM‐3, in the xenogeneic pig‐to‐primate context
Authors: Vadori Marta , Seveso Michela , Besenzon Federica , Bosio Erika , Tognato Elena , Fante Fabio , Boldrin Massimo , Gavasso Sabrina , Ravarotto Licia , Mann Brian E. , Simioni Paolo , Ancona Ermanno , Motterlini Roberto , Cozzi Emanuele
Published: April 2009

Abstract: Background:  Carbon monoxide (CO) interferes with inflammatory and apoptotic processes associated with ischemia–reperfusion injury and graft rejection. Here, the in vitro effects of carbon monoxide releasing molecule‐3 (CORM‐3), a novel water‐soluble carbonyl CO carrier, have been investigated on porcine aortic endothelial cells (PAEC) and primate peripheral blood mononuclear cells (PBMC). Furthermore, the pharmacodynamics and pharmacotolerance of CORM‐3 after administration of single and multiple doses in the primate have been assessed in view of its potential application in pig‐to‐primate xenotransplantation models.

Methods:  For in vitro studies, PAEC and primate PBMC were exposed for 24, 48 and 72 h to CORM‐3 (20 to 1000 μm) and viability was measured using an MTS assay. PAEC and primate PBMC proliferation after exposure to CORM‐3 was assessed by CFSE labelling. Proliferation of primate PBMC against irradiated pig lymphocytes was also assessed. Tumor necrosis factor alpha (TNF‐α) production and Caspase‐3 and ‐7 activity in Concanavalin A (conA)‐stimulated primate PBMC were measured following treatment with CORM‐3. In vivo, CORM‐3 was administered i.v. to cynomolgus monkeys at 4 mg/kg, as single or multiple doses for up to 30 days. The effect of CORM‐3 was evaluated by the assessment of production of TNF‐α and interleukin 1β following PBMC stimulation with LPS by species‐specific ELISA. Complete hematologic and biochemical analyses were routinely performed in treated primates.

Results:  At concentrations <500 μm, CORM‐3 did not alter the viability of PAEC or primate PBMC cultures in vitro, nor did it induce significant levels of apoptosis or necrosis. Interestingly, at concentrations of 300 and 500 μm, significant PAEC proliferation was observed, whilst concentrations ≥50 μm inhibited conA‐activated primate lymphocyte proliferation (IC50 of 345.8 ± 51.9 μm) and the primate xenogeneic response against pig PBMC. Such responses were demonstrated to be CO‐dependent. In addition, CORM‐3 significantly inhibited caspase‐3 and ‐7 activity at concentrations between 200 and 500 μm and caused a significant reduction in TNF‐α production (IC50 332.8 ± 33.9 μm). In vivo, following the administration of multiple doses, TNF‐α production was significantly reduced in comparison to pre‐treatment responses, with decreased levels maintained throughout the study. Moreover, a slight and transient increase in transaminases and bilirubin was observed in animals exposed to multiple doses of CORM‐3.

Conclusions:  These studies suggest that CORM‐3 has anti‐inflammatory and immunomodulatory properties in primates that may result in clinical benefit to allo‐ and xenografted organs.
Dieulafoy’s disease treated by endoscopic haemostatic clipping
Authors: TEO EK , FOCK KM
Published: April 2009

Dieulafoy’s disease is an uncommon cause of upper gastrointestinal haemorrhage. We report three patients with Dieulafoy’s disease treated with the endoscopic haemostatic clip application. There was no recurrent bleeding from the upper gastrointestinal tract after application of the clips in all three patients. Two patients survived while the third succumbed to renal failure after hypovolaemic shock. We conclude that endoscopic haemostatic clip may have a role in the treatment of Dieulafoy’s disease.
The inﬂuence of iron on chronic hepatitis C
Authors: BURT MICHAEL J , COOKSLEY W GRAHAM
Published: April 2009

It has recently been suggested that the hepatic iron concentration can be used to predict the response to interferon in patients with chronic hepatitis C. An hepatic iron concentration greater than 1100 μg/g appears to identify a group of patients that are unlikely to respond to alpha‐interferon. It is not known whether this relationship can be explained by associated variables such as age, gender or disease severity or whether the hepatic iron concentration itself inﬂuences the response to interferon. Furthermore, the hepatic iron concentration is of no value in discriminating responders from non‐responders in patients with hepatic iron concentrations less than 1100 μg/g. The possibility of improving response rates to interferon by pretreatment venesection needs to be explored but currently only limited data are available. Venesection results in a signiﬁcant fall in the serum transaminases but the preliminary results regarding the efﬁcacy of subsequent interferon therapy are unclear. Until the results of prospective controlled trials are available it is concluded that the available evidence does not support venesection before interferon therapy for chronic hepatitis C.
Preliminary report of a randomized, double‐blind placebo‐controlled trial of a Chinese herbal medicine preparation CH‐100 in the treatment of chronic hepatitis C
Authors: BATEY RG , BENSOUSSAN A , FAN YANG YI , BOLLIPO S , HOSSAIN MA
Published: April 2009

The treatment of chronic hepatitis C is relatively unsatisfactory and many patients have turned to unproven alternative medicines to modify the course of their illness. We report a study of a Chinese herbal medicine preparation CH‐100 in the management of chronic hepatitis C. Patients with documented chronic hepatitis C were randomly allocated to receive active herbal or placebo tablets (five tablets thrice daily). Patients were followed monthly and evaluated by a Western and a traditional Chinese medical practitioner. Therapy was monitored by measurement of liver function tests, creatinine and full blood count on a monthly basis. Twenty patients in each group were well matched for age, sex, duration of illness, previous interferon therapy and alcohol intake. Active Chinese herbal medication was associated with a signiﬁcant reduction in alanine aminotransferase (ALT) levels over the 6 month study period (P<0.03). No patient cleared the virus but four normalized their ALT on treatment. Appropriately prescribed Chinese herbal medicine may have a role in the management of chronic hepatitis C and further controlled studies are indicated.
Cryoglobulinaemia among maintenance haemodialysis patients and its relation to hepatitis C infection
Authors: OKUDA KUNIO , YOKOSUKA OSAMU , OTAKE YOSHIO , HAYASHI HARUYUKI , YOKOZEKI KAZUO , KASHIMA TAKASHI , KOBAYASHI SUSUMU , SAKUMA KOSHI , OHNI TSUNEHISA , IRIE YASUBUMI
Published: April 2009

It has been shown that hepatitis C virus (HCV) infection is closely associated with mixed type cryoglobulinaemia. It is also known that HCV infection is rampant among chronic haemodialysis patients. We studied 531 renal failure patients on maintenance dialysis including 170 with positive HCV antibodies for cryoglobulinaemia, and its incidence was compared with controls which consisted of 242 chronic hepatitis C patients without renal failure and 183 healthy adults. Cryoglobulinaemia was present in 30.6% of dialysis patients with HCV infection, 10.8% of dialysis patients without HCV infection, 29.8% of patients with chronic hepatitis C without renal failure, and 0% of healthy adults. Among the 30 new renal failure patients who were started on dialysis within 6 months, four were positive for HCV antibodies, and one of them had cryoglobulinaemia; of the 26 HCV‐negative patients, four (15%) were cryoglobulinaemic. The cryocrit values among dialysis patients were much lower than those of the control cases and other reports on non‐dialysis cases. Patients with cryoglobulinaemia were generally younger compared with patients negative for this condition. There was no correlation between cryoglobulinaemia and past blood transfusion, underlying disease or length of dialysis. Cryoglobulinaemic patients seem to develop renal failure at relatively young ages and a considerable proportion of cryoglobulinaemic dialysis patients may have already had cryoglobulinaemia at the time of the start of haemodialysis. There was no indication that the presence of cryoglobulin in serum adversely affects the liver disease nor increases serum virus load in HCV‐infected dialysis patients. Thus, it was concluded that although HCV infection has a certain role in the development of cryoglobulinaemia in dialysis patients, they develop cryoglobulinaemia less frequently and produce cryoglobulin to a lesser degree in the presence of HCV infection as compared with non‐dialysis patients.
High prevalence of hepatitis G virus infection in multiply transfused children with thalassaemia
Authors: POOVORAWAN YONG , THEAMBOONLERS APIRADEE , CHONGSRISAWAT VORANAUSH , SEKSARN PANYA , SIMMONDS LISA JARVIS† AND PETER
Published: April 2009

We investigated the prevalence of hepatitis G virus (HGV) RNA in relation to the frequency of blood transfusions in thalassaemic children and in volunteer blood donors in Thailand. Furthermore, we studied the frequency of coinfection with hepatitis B virus (HBV) and/or hepatitis C virus (HCV) as well as a possible relationship to the alanine aminotransferase (ALT) status of the blood samples, taken at random from thalassaemic children who have received multiple blood transfusions and from volunteer blood donors. The results show detectable HGV‐RNA in 32.6% of transfusion patients and in 5% of blood donors. The prevalence of HGV‐RNA peaked between the 11th and 50th transfusion. The relationship between HGV infection and ALT status was not statistically relevant.
Autoantibodies against a 210kDa glycoprotein of the nuclear pore complex as a prognostic marker in patients with primary biliary cirrhosis
Authors: ITOH SHINICHI , ICHIDA TAKAFUMI , YOSHIDA TOSHIAKI , HAYAKAWA AKIHITO , UCHIDA MORIAKI , TASHIRO‐ITOH TOMOKO , MATSUDA YASUNOBU , ISHIHARA KIYOSHI , ASAKURA HITOSHI
Published: April 2009

It has been reported that the presence of anti‐nuclear antibody against a 210kDa glycoprotein of nuclear pore complex (anti‐gp210) is highly speciﬁc for primary biliary cirrhosis (PBC). The aim of the present study was to investigate the signiﬁcance of anti‐gp210, especially as a prognostic marker. The presence of anti‐gp210 was ascertained in 113 patients with PBC and 162 controls by indirect immunoﬂuorescence assay using HepG2 cells and immunoblotting analysis using nuclear extracts from HeLa cells. Anti‐gp210 was detected in 25 of the 113 (22.1%) patients. None of the 162 controls was positive for anti‐gp210. The appearance and titre of anti‐gp210 in the patients with PBC did not vary from the time of diagnosis and through their clinical course. Anti‐mitochondrial antibodies (AMA), including antibodies against pyruvate dehydrogenase complex, branched chain α‐ketoacid dehydrogenase complex and α‐ketoglutarate dehydrogenase complex, were not detected by enzyme‐linked immunosorbent assay in ﬁve of the 113 (4.4%) patients with PBC. However, anti‐gp210 alone was positive in one of these ﬁve patients. The difference in prognosis was statistically signiﬁcant; patients with PBC positive for anti‐gp210 died from hepatic failure more frequently than those who were negative (P < 0.01), although there were no statistically signiﬁcant differences in the frequency of jaundice and the histological stage at the time of diagnosis between the two groups. We suggest that the presence of anti‐gp210 is one of the independent prognostic markers able to predict, at the time of diagnosis, a poor outcome in patients with PBC.
Haemodynamic effects of chronic octreotide and tetrandrine administration in portal hypertensive rats
Authors: HUANG YI‐TSAU , CHENG YUH‐REN , LIN HAN‐CHIEH , HONG SU‐MEI CHEN† AND CHUANG‐YE
Published: April 2009

Octreotide is an effective portal hypotensive drug in the control of variceal bleeding. Tetrandrine is a type of calcium channel blocker recently reported to reduce portal hypertension. The present study was undertaken to investigate the haemodynamic effects of octreotide and tetrandrine, alone and in combination, in portal hypertensive rats. Portal hypertension was induced by partial portal vein ligation. Portal hypertensive rats were allocated into one of the four groups: vehicle group (saline, 0.5 mL/day), octreotide group (100 μg/kg per 12 h), tetrandrine group (20 mg/kg per 12 h), and octreotide (100 μg/kg per 12 h) plus tetrandrine (20 mg/kg per 12 h) group. Tetrandrine or saline was administered by gavage, and octreotide by subcutaneous injection. The drug was given for 8 consecutive days, starting 1 day before ligation and continuing onwards. Haemodynamic parameters were measured thereafter, using the radioactive microsphere method. The portal venous pressure and portal tributary blood ﬂow were signiﬁcantly reduced, while portal territory and renal vascular resistances were signiﬁcantly enhanced, by octreotide, tetrandrine, or octreotide plus tetrandrine in portal hypertensive rats, compared with the vehicle group. Our results showed that long‐term administration of octreotide, tetrandrine, or octreotide plus tetrandrine led to portal hypotensive effects in portal hypertensive rats, but octreotide alone exerted better anti‐hyperdynamic effects compared with tetrandrine alone. A combination of octreotide and tetrandrine offered no major beneﬁcial anti‐hyperdynamic effects compared with octreotide alone.
Ki‐67 expression as a prognostic marker in patients with hepatocellular carcinoma
Authors: KING KUANG‐LIANG , HWANG JENG‐JONG , CHAU GAY‐YANG , TSAY SHYH‐HAW , CHI CHIN‐WEN , LEE TUN‐GWE , WU LI‐HWA , WU CHEW‐WUN , LUI WING‐YIU
Published: April 2009

Ki‐67 expression in tumours has been shown to be associated with prognosis in patients with hepatocellular carcinoma (HCC). In this study, primary HCC samples were obtained from 67 patients undergoing surgical resection. None of these patients had been subjected previously to any other form of therapy, such as arterial embolization or chemotherapy. Histologically normal liver tissues from liver resection for metastatic colon cancer were taken as controls (n= 8). Monoclonal antibody against Ki‐67 was used for immunostaining and ﬂow cytometry was used to measure tumour DNA ploidy. The mean Ki‐67 labelling index (percentage of Ki‐67‐positive cells) of the HCC (26 ± 22%; range 0.1–89%) was signiﬁcantly higher than that of the normal controls (39 ± 0.8%, P < 0.05). The mean Ki‐67 labelling index (19 ± 15%; n= 28) of the tumours with diploid DNA pattern was signiﬁcantly lower than those with aneuploid DNA pattern (32 ± 25%, n= 39; P= 0.01). Hepatocellular carcinoma patients (n= 47) with Ki‐67 index >10% had a signiﬁcantly lower disease‐free and overall survival than those (n= 20) with Ki‐67 index ≤10% (P= 0.0009 and P= 0.02, respectively). Multivariate analysis showed that Ki‐67 expression and tumour node metastasis stage were two independent prognostic factors for disease‐free and overall survival rates. Our results suggest that the expression of Ki‐67 is an independent prognostic indicator for patients with HCC after resection and could be of assistance in the decision‐making of adjuvant therapy.
Economic evaluation of Helicobacter pylori eradication for the treatment of duodenal ulcer disease in Japan: A decision analysis to assess eradication strategy in comparison with a conventional strategy
Authors: HABU YASUKI , INOKUCHI HIDETO , KIYOTA KEISUKE , HAYASHI KYOHEI , WATANABE YOSHIYUKI , KAWAI KEIICHI , STÅLHAMMAR NILS‐OLOV
Published: April 2009

To evaluate the socio‐economic effects of Helicobacter pylori eradication in the treatment of duodenal ulcer disease in Japan, a clinical decision analysis was performed to assess H. pylori eradication therapy compared with the conventional strategy of maintenance with histamine‐2 receptor antagonists. A decision tree‐based state transition model (Markov chain approach) implemented to simulate a 5 year period of follow up was constructed. The H. pylori eradication strategy was found to be superior to the conventional maintenance strategy with regard to clinical effectiveness and other dimensions of a patient’s outcome. Furthermore, in a long‐term perspective, the eradication strategy was less costly than the maintenance strategy. Helicobacter pylori eradication should be recommended as the ﬁrst choice treatment of H. pylori‐positive duodenal ulcer patients. The clinical implication of H. pylori eradication entails an improvement in clinical effectiveness and other dimensions of a patient’s outcome and a signiﬁcant reduction in the costs of duodenal ulcer treatment. The long‐term total costs do not depend on the initial drug cost of an eradication regimen. Pursuing a high eradication rate of H. pylori is essential in improving the patient’s outcome and the cost‐effectiveness of treatment.
Low dose of clarithromycin in triple therapy for the eradication of Helicobacter pylori: One or two weeks?
Authors: BO’ N DAL , MARIO F DI , BATTAGLIA G , BUDA A , LEANDRO G , VIANELLO F , KUSSTATSCHER S , SALANDIN S , PILOTTO A , CASSARO M , S VIGNERI  , RUGGE M
Published: April 2009

The aims of this pilot study were: (i) to compare the efﬁcacy of low‐dose clarithromycin (250 mg twice daily) for 1 or 2 weeks; and (ii) to evaluate possible therapeutic advantages in associating the low‐dose clarithromycin with an anti‐secretory agent or tripotassium dicitrate bismuthate (De Nol; Yamanouchi Pharm, Corugate Milano, Italy). A prospective, randomized, open trial was carried out on consecutive outpatients with dyspeptic symptoms and Helicobacter pylori infection. We enrolled 129 patients in one of the following schedules: (A) De Nol 120 mg q.i.d., clarithromycin 250 mg b.i.d. and metronidazole 250 mg q.i.d. for 2 weeks; (B) omeprazole 20 mg b.i.d., clarithromycin 250 mg b.i.d. and metronidazole 250 mg q.i.d. for 2 weeks; or (C) omeprazole 20 mg b.i.d., clarithromycin 250 mg b.i.d. and metronidazole 250 mg q.i.d. for 1 week. Results were evaluated by Per Protocol (PP) and Intention‐To‐Treat analysis (ITT). Eradication rate was 100% after treatment A, 92.6% after treatment B and 86.5% after treatment C by PP and 83.3, 75.7, and 68.1%, respectively by ITT. Side effects were reported by 16 subjects: 26.6% in group A; 9.1% in group B; and 7.5% in group C; in two cases side effects led to the withdrawal of the treatment. In conclusion, 500 mg clarithromycin per day in association with omeprazole and metronidazole, for 1 week gave comparable results to the same schedule for a 2 week period. The use of clarithromycin with bismuth and metronidazole produced a therapeutic gain compared with both of the anti‐secretory schedules, although this was not statistically signiﬁcant.
A prospective randomized study of amoxycillin and omeprazole with and without metronidazole in the eradication treatment of Helicobacter pylori
Authors: KOIZUMI WASABURO , TANABE SATOSHI , HIBI KENICHI , IMAIZUMI HIROSHI , OHIDA MASAHITO , OKABE HARUYA , SAIGENJI KATSUNORI , OKAYASU ISAO
Published: April 2009

A combination of amoxycillin and omeprazole is often used to treat Helicobacter pylori infection. A three‐drug regimen comprising metronidazole, amoxycillin and omeprazole has been proposed as an alternative therapy. In a prospective, randomized, comparative study, we evaluated these two regimens with respect to safety and efﬁcacy in patients with H. pylori infection. Sixty patients with peptic ulcer (gastric, 32 patients; duodenal, 28 patients) who had a history of ulcer recurrence were randomly assigned to dual therapy with amoxycillin (500 mg three times daily for 2 weeks) and omeprazole (20 mg once daily for 8 weeks) or to triple therapy with metronidazole (500 mg twice daily for 2 weeks) plus amoxycillin and omeprazole, given in the same dosages as dual therapy. Forty‐eight patients completed the protocol; treatment was discontinued because of side effects in nine patients, and three patients dropped out of the study. On the basis of all patients treated, the rate of H. pylori eradication was signiﬁcantly higher for triple therapy 20/23 cases, 87.0%; 95% conﬁdence interval (CI), 0.664–0.972) than for dual therapy 13/25, 52.0%; 0.313–0.722; P < 0.05). On an intention‐to‐treat basis, the difference between the groups in the rate of H. pylori eradication was marginally signiﬁcant (P= 0.06 [0.028–0.512]). Side effects were reported by ﬁve patients receiving triple therapy (skin rash, one; nausea, two; headache, one; abdominal pain, one), and four patients receiving dual therapy (skin rash, two; abdominal pain, one; diarrhoea, one). All side effects resolved spontaneously after termination of treatment. There was no signiﬁcant difference in safety between the two regimens. Triple therapy with metronidazole, amoxycillin, and omeprazole was signiﬁcantly more effective for the eradication of H. pylori than dual therapy with amoxycillin and omeprazole alone. The safety of these regimens was similar, and triple therapy was found to be clinically acceptable.