bluuebunny/crossref_metadata_2025 huggingface dataset:
The use of clip cages to restrain insects reduces leaf expansion systemically in Rumex obtusifolius
Authors: Moore J. P. , Taylor J. E. , Paul N. D. , Whittaker J. B.
Published: December 2004

Abstract. 1. Clip cages have been used widely by experimental ecologists to contain insects on plants.

2. Under controlled conditions, the effect of applying clip cages alone and clip cages and the chrysomelid beetle Gastrophysa viridula on systemic leaf expansion of Rumex obtusifolius was investigated. Treatments were applied to the fully expanded fourth leaf and expansion of leaf 8 was measured over a period of 22 days.

3. The application of clip cages reduced the rate at which leaf area increased and led to reductions in final leaf areas.

4. Clip cages have systemic effects on plant development and these effects are sustained even after the clip cage is removed. Investigators should take this into account in designing experiments.
The significance of a facultative bacterium to natural populations of the pea aphid Acyrthosiphon pisum
Authors: Darby A. C. , Tosh C. R. , Walters K. F. A. , Douglas A. E.
Published: December 2004

Abstract. 1. 
Laboratory studies have implicated various accessory bacteria of aphids as important determinants of aphid performance, especially on certain plant species and under certain thermal regimes. One of these accessory bacteria is PABS (also known as T‐type), which is distributed widely but is not universal in natural populations of the pea aphid
Acyrthosiphon pisum in the U.K.

2.  To explore the impact of PABS on the performance of A. pisum
, the nymphal development time and fecundity of aphids collected directly from natural populations and caged on the host plant
Vicia faba in the field were quantified. Over 4 consecutive months June–September 1999, the performance of PABS‐positive and PABS‐negative aphids did not differ significantly.

3.  Deterministic modelling of the performance data showed that the variation in simulated population increase of PABS‐positive and PABS‐negative aphids would overlap substantially.

4.  Analysis of aphids colonising five host plants ( Lathyrus odoratus
,
Medicago sativa
,
Pisum sativum
,
Trifolium pratense
,
Vicia faba ) between April and September 2000 and 2001, identified no robust differences between the distribution of PABS‐positive and PABS‐negative aphids on different plants and with season or temperature.

5.  It is concluded that PABS is not an important factor shaping the performance or plant range of A. pisum under the field conditions tested. Reasons for the discrepancies between this study and laboratory‐based studies are considered.
Dispersal patterns of an Australian gall‐forming thrips and its host tree (Oncothrips tepperiandAcacia oswaldii )
Authors: Mcleish Michael J. , Perry S. P. , Gruber D. , Chapman T. W.
Published: December 2004

Abstract.1. Clark and Evans' nearest‐neighbour method was used to evaluate the distribution pattern ofAcacia oswaldiiandOncothrips tepperipopulations.

2. A random distribution pattern was supported statistically for both the host trees and trees that were parasitised byO. tepperi.

3. It is therefore likely that dispersal distances of colonising thrips are large enough to be able to traverse the 1 km2study site.
Contrasting exotic Solenopsis invicta and native Forelius pruinosus ants as mutualists with Catalpa bignonioides, a native plant
Authors: Ness J. H.
Published: December 2004

Abstract. 1. The suitability of the red imported fire ant Solenopsis invicta Buren and a native ant Forelius pruinosus (Roger) as participants in a food‐for‐protection mutualism with a native nectaried tree Catalpa bignonioides Walter was compared.

2. The mean mortality of folivore larvae of Ceratomia catalpae Boisduval was similar for S. invicta and F. pruinosus although S. invicta attacked fewer caterpillar aggregations and was a devastating pupal predator. Solenopsis invicta also differed from the native ant in that it attacked the parasitoid Cotesia congregata Say, another plant mutualist, and visited extrafloral nectaries less frequently.

3. Habitats invaded by S. invicta are characterised by a scarcity of both herbivores and of beneficial insects that visit extrafloral nectaries. The plants do not require protection, and extrafloral nectaries are visited rarely. Although plants are defended incidentally by S. invicta, the insect‐plant mutualism therein is greatly simplified or defunct.
Measuring dispersal and detecting departures from a random walk model in a grasshopper hybrid zone
Authors: Bailey R. I. , Lineham M. E. , Thomas C. D. , Butlin R. K.
Published: December 2004

Abstract.1. The grasshopper speciesChorthippus brunneusandC. jacobsiform a complex mosaic hybrid zone in northern Spain. Two mark–release–recapture studies were carried out near the centre of the zone in order to make direct estimates of lifetime dispersal.

2. A model framework based on a simple random walk in homogeneous habitat was extended to include the estimation of philopatry and flying propensity. Each model was compared with the real data, correcting for spatial and temporal biases in the data sets.

3. All four data sets (males and females at each site) deviated significantly from a random walk. Three of the data sets showed strong philopatry and three had a long dispersal tail, indicating a low propensity to move further than predicted by the random walk model.

4. Neighbourhood size estimates were 76 and 227 for the two sites. These estimates may underestimate effective population size, which could be increased by the long tail to the dispersal function. The random walk model overestimates lifetime dispersal and hence the minimum spatial scale of adaptation.

5. Best estimates of lifetime dispersal distance of 7–33 m per generation were considerably lower than a previous indirect estimate of 1344 m per generation. This discrepancy could be influenced by prezygotic isolation, an inherent by‐product of mosaic hybrid zone structure.
Parental effects inPieris rapaein response to variation in food quality: adaptive plasticity across generations?
Authors: Rotem Karin , Agrawal Anurag A. , Kott Laima
Published: December 2004

1. Herbivores using seasonal resources must cope with variation in the quality of their host plants. The effects of variation in protein concentration of artificial diet and glucosinolate concentration in canola,Brassica napus, onPieris rapaeparental and progeny growth were investigated.

2. The hypothesis that parents respond to variation in food quality by altering the phenotype of their progeny to enhance progeny fitness was tested. Consistent with previous studies,P. rapaewas not affected strongly by variation in the protein concentration of artificial diet and had equal mass on completing development.

3. The mass of individual eggs ofP. rapaeprogeny was correlated negatively with the amount of protein in the diet on which parents fed. Moreover, mothers reared in extreme conditions (high and low protein) produced progeny that grew best under those conditions. These potentially adaptive parental effects were detected early in progeny growth but not later in their development.

4. Early larval growth ofP. rapaewas affected negatively by increasing glucosinolates inB. napusplants, although no effects of glucosinolates were detected later in growth or on the progeny's phenotype.

5. Thus, evidence is presented that variation in food quality (protein concentration) has major consequences for the progeny ofP. rapae. Given the multivoltine life history ofP. rapaeand the seasonal differences in food quality it encounters, such parental effects may be adaptive.
Time to progression is dependent on the expression of the tumour suppressor PTEN in ovarian cancer patients
Authors: Schöndorf T. , Göhring U.‐J. , Roth G. , Middel I. , Becker M. , Moser N. , Valter M. M. , Hoopmann M.
Published: December 2004

Background Quantitative analyses of PTEN expression of ovarian cancer tissues were performed in this study. PTEN expression was investigated in terms of each patient's progression‐free interval to indicate the role of PTEN in the generation of platinum refractory tumours.

Materials and methods The study group comprised 20 ovarian cancer patients from whom fresh frozen tissues of both the primary tumour and specimens of progressive disease were available. The PTEN protein and phosphorylation of the downstream effector proteinkinase B (PKB) were quantified by Western blot analyses and subsequent densitometry. Data were analyzed for individual PTEN variation with respect to the clinical course as defined by the progression‐free interval.

Results Applying the usual clinical criteria for platinum‐sensitivity after progression, seven patients were considered platinum‐sensitive whereas 13 patients had suffered a progression within 12 months after the chemotherapy. In 5/7 (71%) cases with prolonged time to progression, an increase in PTEN was observed. Decline of PTEN expression occurred in 9/13 (69%) patients with poor outcome. PTEN expression corresponds inversely to PKB phosphorylation in 14/20 (70%) tissues investigated.

Conclusions The data suggest that decreased PTEN expression accompanies the progression of ovarian cancer. Declining PTEN expression results in a shortened relapse‐free interval, whereas an increase of PTEN prolongs the time to progression. However, as far as recurrence occurs, PTEN is not the only mechanism to suppress tumour progression in ovarian cancer.
Cardiovascular responses to endurance training in children: effect of gender
Authors: Obert P. , Mandigouts S. , Nottin S. , Vinet A. , N’Guyen L. D. , Lecoq A. M.
Published: December 2004

BackgroundThe aim of the present study was to determine in healthy children the effect of a well‐controlled endurance training programme on cardiac function at maximal exercise and to define whether gender affects the training‐induced cardiovascular response. The contribution of factors potentially involved in those adaptations such as cardiac dimensions and diastolic and systolic function was also investigated.

MethodsThirty‐five l0–11‐year‐old children took part in this study: 19 children (10 girls and nine boys) were assigned to participate in a 13‐week endurance training programme (3 × 1 h week−1, intensity: > 80% HR max), and 16 (seven girls and nine boys) served as a control group. A resting echocardiographic evaluation and a maximal upright cycle test, including measurement of stroke volume (SV), cardiac output (Q) and blood pressure, were performed in all children before and after the study period.

ResultsThe training programme led to a rise in maximal O2uptake (V̇O2max), brought about however, only by an increase in SVmaxin both genders. Moreover, the boys increased their V̇O2maxto a greater extent than the girls (boys: +15%; girls: +8%) only because of a higher SVmaximprovement (boys: +15%; girls: +11%). No alterations were noticed in the SV pattern from rest to maximal exercise, indicating that the increase in SVrestwas a key factor in the improvement of SVmaxand thus V̇O2max. Regarding resting echocardiographic data, an increase in the left ventricular end‐diastolic diameter, concomitant with an improvement in diastolic function, was observed after training and constituted an essential element in the rise in V̇O2maxafter training in these children. Moreover, during maximal exercise, a decrease in systemic vascular resistances, probably indicating peripheral cardiovascular adaptive changes, might also play an important role in the increase in V̇O2max.

Conclusion

Whatever gender, aerobic training increases V̇O2maxin children, mediated by an improvement in SVmaxonly. Similar mechanisms, including loading conditions and cardiac morphology, seem to be involved in both genders in order to explain such an improvement.
Decreasing peripheral resistance during pregnancy monitored at the cubital artery
Authors: Schiessl B. , Strasburger C. J. , Spannagl M. , Kainer F.
Published: December 2004

Background During the physiological course of pregnancy maternal peripheral vascular resistance decreases. Using the Doppler technique, the decreasing resistance and disappearance of the early diastolic notch in the arterial uterine perfusion can be observed. These changes in the Doppler flow velocity wave form are caused by the trophoblast invasion in which the spiral arteries loose their musculoelastic structure of the arterial wall. The aim of this study was to investigate whether maternal systemic alteration in the Doppler flow velocity waveform and Pulsatility Index (PI) could be monitored at the cubital artery during normal pregnancy.

Methods and results Forty‐five patients were investigated in a prospective study in 4–6‐week intervals for Dopplersonographic evaluation of the two uterine arteries and the right cubital artery; 20 of the patients were also investigated after delivery. The presence or absence of the early diastolic reverse flow of the cubital artery was analyzed. Furthermore, the PI was calculated for all vessels. With increasing duration of gestation, the reverse flow of the cubital artery disappeared. During the course of gestation a significant decrease in the PI of the cubital artery was observed in addition to a significantly decreased PI of the uterine arteries. The PI of the placental and nonplacental uterine arteries showed a significant correlation to the PI of the cubital artery.

Conclusions This study demonstrates systemical changes in arterial resistance correlating to those of the uterine arteries during the course of physiological pregnancy.
Associations of metabolic variables with arterial stiffness in type 2 diabetes mellitus: focus on insulin sensitivity and postprandial triglyceridaemia
Authors: Van Dijk R. A. J. M. , Bakker S. J. L. , Scheffer P. G. , Heine R. J. , Stehouwer C. D. A.
Published: December 2004

Background Type 2 diabetes mellitus is associated with an increased risk of atherothrombotic disease, which may in part be mediated through increased arterial stiffness. We investigated to what extent increased arterial stiffness is associated with cardiovascular risk factors that commonly cluster in type 2 diabetes.

Design In this cross‐sectional, observational study we assessed, in 81 subjects with type 2 diabetes, local distensibility and compliance of the common carotid and femoral arteries, and carotido‐femoral transit time. We subsequently investigated whether these measures of arterial stiffness were determined by diabetes duration, blood pressure, lipid levels (including fasting and postprandial triglyceridaemia), hyperglycaemia, BMI, waist‐to‐hip ratio, insulin resistance (as measured by a euglycaemic, hyperinsulinaemic clamp), hyperinsulinaemia, cigarette smoking, or the use of alcohol.

Results In multivariate analysis, insulin‐mediated glucose uptake was positively [standardized β, 0·21 (P = 0·05)] associated with the carotid artery compliance coefficient, and the use of alcohol was negatively associated with the femoral artery compliance coefficient [standardized β, –0·25 (P = 0·03)]. Except for mean arterial pressure, which was negatively associated with all outcome variables, the other cardiovascular risk factors that we investigated were not significantly associated with the distensibility coefficients of both the carotid and femoral artery, or with the carotido‐femoral transit time.

Conclusions In subjects with type 2 diabetes mellitus, insulin resistance and the use of alcohol were associated with increased arterial stiffness, which supports the hypothesis that increased arterial stiffness can act as a mediating factor in the association between type 2 diabetes mellitus and increased risk of atherothrombotic disease. We found no evidence for an association between fasting or postprandial triglyceridaemia and arterial stiffness.
Endothelial vasodilatory function is related to the proportions of saturated fatty acids and alpha‐linolenic acid in young men, but not in women
Authors: Steer P. , Vessby B. , Lind L.
Published: December 2004

Background Fatty acid composition of serum lipids is associated with cardiovascular disease. As attenuated endothelium‐dependent vasodilation (EDV) is an early event in atherosclerosis, we investigated the relationships between endothelial vasodilatory function and the proportion of serum fatty acids, reflecting dietary fat quality, in 74 healthy men and women, aged 20–30 years.

Design Endothelium‐dependent vasodilation and endothelium‐independent vasodilation (EIDV) was studied in the forearm during local administration of methacholine (2 and 4 µg min−1) and nitroprusside (5 and 10 µg min−1). Forearm blood flow was determined with venous occlusion plethysmography. An endothelial function index was calculated as the EDV/EIDV ratio.

Results The endothelial function index was inversely related to the total proportion of saturated fatty acids (r = −0·41, P < 0·05), in particular lauric and myristic acid (r = −0·37 and r = −0·36, respectively, P < 0·05 for both), and was positively related to the proportion of alpha‐linolenic acid (r = 0·45, P < 0·01) in men only. Total serum nonesterified fatty acid (NEFA) concentration was not significantly related to endothelial vasodilatory function. By multiple stepwise regression analysis, including age, blood pressure, body mass index, and serum cholesterol, triglyceride and NEFA as confounders, myristic acid and alpha‐linolenic acid were independent predictors of the endothelial function index in men only (r = −0·39 and r = 0·47, respectively, P < 0·01 for both).

Conclusions We conclude that serum fatty acid composition predicts endothelial vasodilatory dysfunction independently of serum NEFA and cholesterol levels in young, healthy men in their third decade of life, whereas fatty acid composition seems to be less important in women at this age. As a result of the large number of analyses performed, these findings need to be verified by other studies.
Measles virus induces apoptosis in uninfected bystander T cells and leads to granzyme B and caspase activation in peripheral blood mononuclear cell cultures
Authors: Vuorinen T. , Peri P. , Vainionpää R.
Published: December 2004

Background Measles causes lymphopenia and depresses cell‐mediated immunity, but the mechanisms of immunosuppression and cell loss are poorly known.

Methods We have used an in vitro model of measles virus (MV)‐infected peripheral blood mononuclear cells (PBMCs) and phytohaemagglutinin‐stimulated PBMCs in order to assess MV–leucocyte interactions. Cell population undergoing apoptosis was measured by flow cytometry and Annexin‐V‐fluos staining. The expression of Fas, FasL, TNRF1, and Bcl‐2 was analyzed by flow cytometry and Western blotting, and activation of caspase cascade was measured using a colourimetric caspase substrate set. The effects of caspase inhibitors were detected by flow cytometry.

Results Measles virus was able to infect monocytes, but interestingly induced apoptosis in uninfected T cells, indicating that induction of apoptosis in T cells is mediated by MV‐infected adherent cells. Only 1% of T cells contained MV antigen day 3 p.i. Interestingly the percentage of early apoptotic T cells at the same time was 35%, showing that apoptosis was not the result of MV infection in T cells. Measles virus‐induced Fas but not FasL or TNFR1 expression on PMBC, as well as activation of granzyme B and caspase cascade. Simultaneously, overexpression of Bcl‐2 protein was detected. Caspase inhibitor decreased the amount of apoptotic T cells.

Conclusion Measles virus‐infected monocytes induce apoptosis in uninfected T cells, suggesting that infected monocytes probably interact via cell–surface molecules with uninfected T cells and induce apoptosis by indirect mechanisms. Apoptosis of the lymphocytes may contribute to the pathogenesis of MV‐induced immunosuppression and cell loss.
A modified PCR‐SSP method for the identification of ABO blood group antigens
Authors: Downing J. , Darke C.
Published: December 2004

The ABO blood group antigens are carbohydrate molecules synthesized by the glycosyltransferases encoded by the ABO gene on chromosome 9. Kidney transplantation across the ABO barrier generally leads to rapid humoral graft rejection due to the presence of naturally occurring antibodies to the A and B antigens. We have developed a method for ABO typing our cadaveric organ donors by the polymerase chain reaction using sequence‐specific primers (PCR‐SSP). The method uses 12 primers in eight PCR mixtures and is performed under the same conditions as our routine HLA‐A, B, C PCR‐SSP typing. The PCR‐SSP‐based types of 166 regular blood donors and 148 cadaveric organ donors all showed total concordance with their serologically assigned ABO groups. Six individuals possessing the ABO A subgroups (A3, Ax and Aend) all typed as A1 by PCR‐SSP, as expected. PCR‐SSP is an appropriate method for ABO typing of cadaveric organ donors and, importantly, enables both ABO and HLA typing to be performed on the same DNA material.
Platelet activation in heart transplant recipients
Authors: Hognestad Aina , Michelsen Annika , Brosstad Frank , Damås Jan K , Holm Torbjørn , Simonsen Svein , Kjekshus John K , Aukrust Pål , Andreassen Arne K
Published: December 2004

Abstract:  An inappropriate and persistent immune activation has been suggested to contribute to long‐term mortality and morbidity after heart transplantation. Several lines of evidence suggest that platelets do not only promote thrombus formation, but also act as inflammatory cells. In the present study, we investigated if long‐time survivors of heart transplantation (mean time since transplantation 6.5 yr) were characterized by enhanced platelet activation as assessed by different experimental approaches. Our main findings when comparing heart transplant recipients (n = 52) and age‐ and sex‐matched healthy controls (n = 38) were: (i) platelets from heart transplant recipients showed enhanced expression of both P‐selectin and CD63 as assessed by flow cytometry; (ii) platelets from these patients also contained significantly increased levels of soluble CD40 ligand and tended to release higher levels of this cytokine upon SFLLRN stimulation as assessed by enzyme immunoassay; (iii) heart transplant recipients had increased levels of soluble P‐selectin in platelet‐free plasma; and (iv) the enhanced platelet activation after heart transplantation was most pronounced in those with concomitant hypertension. These findings suggest that long‐term survivors of heart transplantation are characterized by enhanced activation of platelets, possibly contributing to the persistent immune activation and clinical complications in these patients.
The incidence and significance of late acute cellular rejection (>1000 days) after liver transplantation1
Authors: Florman Sander , Schiano Thomas , Kim Leona , Maman Dan , Levay Adam , Gondolesi Gabriel , Fishbein Thomas , Emre Sukru , Schwartz Myron , Miller Charles , Sheiner Patricia
Published: December 2004

Abstract:  Acute cellular rejection (ACR) after liver transplantation occurs in as much as 70% of patients within the first year. There is very little known about ACR that occurs more than 1 yr after transplant, and it is generally believed that late occurring ACR may be more resistant to medical treatment and is associated with a higher rate of chronic ductopenic rejection and graft loss. A total of 532 recipients with more than 1000 d follow‐up and who did not have hepatitis C were identified. Forty‐three (8.1%) had biopsy proven late ACR at a mean of 1545 ± 441 d post‐transplant. Additionally, 38 of the 43 (88.4%) patients with late ACR had earlier episodes of ACR before 1000 d post‐transplant vs. only 295 of the 488 patients (60.5%) that did not have late ACR (p < 0.01). The incidence of primary sclerosing cholangitis (PSC) was 32.6% among patients with late ACR and 11.1% among patients without late ACR (p < 0.01). The overall patient survival for patients who had late ACR (n = 43) is 81.4% while for patients without late ACR (n = 488) it is 82.0% (p = ns). Patients remain at risk for ACR even after 1000 d post‐transplant, particularly those with PSC.
Interstitial alterations in renal cortex in acute tubular necrosis (ATN) post‐renal transplantation and in patients with ATN not related to renal transplant
Authors: Neto Miguel Moyses , Costa Roberto S , Volpini Rildo A. , Garcia Tânia MP , Rodrigues Fernanda F , Coimbra Terezila M
Published: December 2004

Abstract:  Background:  Abnormalities of renal function with long‐term implications can persist after acute tubular necrosis (ATN), probably because of permanent loss of nephrons. Residual areas of fibrosis are also observed in the renal cortex post‐ATN. In this study, we investigate the interstitial alterations post‐ATN using histological and immunohistochemical methods.

Methods:  We studied 11 patients with ATN of different etiologies and 19 patients with ATN post‐renal transplantation. Eleven patients with ATN post‐renal transplantation and one with ATN not related to renal transplantation were submitted to more than one biopsy because of delayed renal function recovery. The immunohistochemical studies were performed using α‐smooth muscle‐actin (α‐SM‐actin), endothelin, nuclear factor‐κB (NF‐κB), Jun‐N‐terminal kinase (p‐JNK) and fibronectin antibodies. We also analyzed the urinary content of transforming growth factor‐β (TGF‐β) during the acute phase of ATN.

Results:  The immunohistochemical studies showed increased α‐SM‐actin, fibronectin, endothelin, p‐JNK and NF‐κB staining in the tubulointerstitium area from the renal cortex of all patients when compared with controls (p < 0.001), and these increase persisted in the patients submitted to sequential biopsies. One of the patients with ATN without renal transplant and six patients with ATN post‐renal transplant developed chronic renal failure. There was a significant increase of TGF‐β excretion in the urine of patients with acute renal failure (p < 0.01) compared with control.

Conclusion:  Our data show that the enhancement of renal TGF‐β production and the persistent increase of myofibroblasts, fibronectin, endothelin, p‐JNK and NF‐κB in renal cortex tubulointerstitium post‐ATN may explain the impaired recovery of renal function observed in patients post‐ATN frequently observed in patients with ATN post‐renal transplant.
Liver transplant for hepatocellular cancer: a treatment for the select few
Authors: Wong Linda L , Tsai Naoky , Limm Whitney , Wong Livingston
Published: December 2004

Abstract:  Hepatocellular cancer (HCC) is increasing in incidence and liver transplant (LT) is likely the best treatment for long‐term survival and decreased recurrence in those with localized disease. Optimal treatment for HCC is limited by advanced stage at presentation and available donors. We retrospectively reviewed 232 cases of HCC from 1993 to 2002 referred to our medical center, which also has the only transplant program in the state. Demographic data, risk factors, stage, previous treatment, candidacy for LT, and outcome were noted. Stage distribution was as follows: stage I – nine patients, II – 115 patients, III – 31 patients, and IV – 76 patients. Mean age was 61.1 yr and 62 patients were over the age of 70 yr. Thirty‐five patients (15.1%) were offered LT evaluation. Nineteen patients (8.2%) eventually underwent LT, five are currently on a transplant waiting list, three were on a list but removed due to death or progression of disease, six were evaluated but not listed, and two refused evaluation. Five patients were transplanted because implementation of the Model of End‐stage Liver Disease (MELD) system for organ allocation. Of 19 patients who underwent LT, 14 are still alive 3–52 months post‐transplant, three developed recurrent HCC, and two died of lung cancer. Although LT may be the best treatment for HCC, a relatively small number of patients (15%) will qualify for this therapy. About half of those who qualify will actually undergo LT. This further emphasizes the need for other strategies such as vaccination, chemoprevention, and early detection to improve survival from HCC.
Modified colorimetric assay for uricase activity and a screen for mutant Bacillus subtilis uricase genes following StEP mutagenesis
Authors: Huang Su‐Hua , Wu Tung‐Kung
Published: December 2004

This study describes a modified colorimetric assay for uricase activity in flexible 96‐well microtiter plates using the uricase/uric acid/horseradish peroxidase/4‐aminoantipyrine/3,5‐dichloro‐2‐hydroxybenzene sulfonate colorimetric reaction. The utility of this assay was demonstrated in a screen for mutant uricase enzymes derived from the uricase gene of the thermophilic bacterium Bacillus subtilis by a modified staggered extension process (StEP) mutagenesis. An Escherichia coli library of StEP‐derived uricase mutant clones was screened yielding two identical active mutant uricase genes. Two motifs conserved in eukaryotic and prokaryotic uricases are highly conserved in the mutant uricase. The mutant uricase protein was found to exhibit high uricase activity (13.1 U·mg−1). Finally, the modified colorimetric method is much more efficient than the conventional ones and greatly reduces assay time from 4 days to less than 20 h.
Isolation and biochemical characterization of two soluble iron(III) reductases from Paracoccus denitrificans
Authors: Mazoch Jiří , Tesařík Radek , Sedláček Vojtěch , Kučera Igor , Turánek Jaroslav
Published: December 2004

Two soluble enzymes (FerA and FerB) catalyzing the reduction of a number of iron(III) complexes by NADH, were purified to near homogeneity from the aerobically grown iron‐limited culture of Paracoccus denitrificans using a combination of anion‐exchange chromatography (Sepharose Q), chromatofocusing (Mono P), and gel permeation chromatography (Superose 12). FerA is a monomer with a molecular mass of 19 kDa, whereas FerB exhibited a molecular mass of about 55 kDa and consists of probably two identical subunits. FerA can be classified as an NADH:flavin oxidoreductase with a sequential reaction mechanism. It requires the addition of FMN or riboflavin for activity on Fe(III) substrates. In these reactions, the apparent substrate specificity of FerA seems to stem exclusively from different chemical reactivities of Fe(III) compounds with the free reduced flavin produced by the enzyme. Observations on reducibility of Fe(III) chelated by vicinal dihydroxy ligands support the view that FerA takes part in releasing iron from the catechol type siderophores synthesized by P. denitrificans. Contrary to FerA, the purified FerB contains a noncovalently bound redox‐active FAD coenzyme, can utilize NADPH in place of NADH, does not reduce free FMN at an appreciable rate, and gives a ping‐pong type kinetic pattern with NADH and Fe(III)‐nitrilotriacetate as substrates. FerB is able to reduce chromate, in agreement with the fact that its N‐terminus bears a homology to the previously described chromate reductase from Pseudomonas putida. Besides this, it also readily reduces quinones like ubiquinone‐0 (Q0) or unsubstituted p‐benzoquinone.
Susceptibility and lesion development in Scots pine saplings infected with Peridermium pini in northern Finland
Authors: Kaitera J.
Published: December 2004

Susceptibility of Pinus sylvestris provenances from northern Finland to Peridermium pini and lesion development caused by the rust were investigated by inoculating 768 saplings. Resin‐top disease symptoms on the stem including stem swelling, occurrence of spermatial fluid and rust aecia, were recorded over a period of 9 years. The disease incidence was low (3.6%), varying within the range of 0–9.4% among pine provenances and 3.4–3.9% among spore sources in 1994–2002. Aecia were produced for the first time 2–4 years after inoculation, appearing mainly in early June, 2–9 years after inoculation. The aecia ruptured in late July to early August, and the overall sporulation lasted for 1–8 years among the pine provenances. Spermatial fluid was observed regularly in the year prior to aecia development 2–7 years after inoculation. Annual fresh swelling was observed for 1–6 years after inoculation. No statistically significant differences occurred neither between the pine provenances, the spore sources nor the years with regard to all the major disease variables. The results suggest that Scots pine provenances from northern Finland are highly resistant to P. pini regardless of the pine provenance or spore source.
Rust resistance in Salix induced by inoculations with avirulent and virulent isolates of Melampsora larici‐epitea
Authors: Pei M. H. , Ruiz C. , Hunter T. , Bayon C.
Published: December 2004

Six Salix clones and four Melampsora larici‐epitea isolates were used in two leaf‐disc inoculation experiments to determine whether disease severity was affected by the presence of both virulent and avirulent rust genotypes. In the first experiment, an equal amount of urediniospores of a virulent isolate and an avirulent isolate was applied simultaneously using four levels of spore suspension. In the second, the willows were inoculated with one isolate first and then, after 3 days, with another using two spore concentrations. In the first experiment, overall rust spore production was reduced by 48.6% in inoculations with mixed inocula compared with that in the inoculations with single virulent isolates. In the second experiment, 20 of the 36 combinations involving pre‐inoculations with avirulent isolates showed significant reduction in spore production. The suppressive effects on rust sporulation were more obvious at the higher inoculum densities and on the clones S. × calodendron and S. × mollissima‘Q83’.
Decay and canker caused byInonotus rickiispreading on more urban tree species
Authors: Annesi T. , Coppola R. , Motta E.
Published: December 2004

Inonotus rickiiwas detected for the first time causing cankers and decay inAcer negundoandCeltis australisin Italy. In a boxelder boulevard, declining trees showed sparse foliage, exudations and cracks in the bark; in some cases, chlamydospore masses were present. Five isolates were collected and compared by growth testsin vitroand electrophoretic analyses; three isolates from the same boulevard showed very similar physiological characters. The increasing importance of the pathogen in urban areas is underlined and discussed.
Field tests on biological control of Heterobasidion annosum by Phaeotheca dimorphospora in comparison with Phlebiopsis gigantea
Authors: Roy G. , Laflamme G. , Bussières G. , Dessureault M.
Published: December 2004

In field tests conducted on freshly cut red pine (Pinus resinosa) logs, the antagonistic fungus Phaeotheca dimorphospora formulated as pre‐colonized rye grains reduced colonization of wood tissue by Heterobasidion annosum to less than 2% over the 2 months following inoculation. However, compared to controls, a water suspension of P. dimorphospora did not reduce the infection, but rather promoted colonization by H. annosum after a 2‐month exposure. Production of antifungal metabolites by P. dimorphospora in the rye grains and better colonization of the logs by the antagonist were observed with the rye grain formulation. In this field test, the most effective treatment to control H. annosum was the application of an oidial suspension of Phlebiopsis gigantea. This fungus completely inhibited colonization by H. annosum mostly by rapid colonization of wood tissue. All treatments with P. gigantea, P. dimorphospora and rye grains influenced the naturally occurring mycoflora. The most important species composing this mycoflora were Hormonema sp., Mortierella spp., Mucor hiemalis f. hiemalis, Nectria sp., Pesotum sp., P. gigantea, Tympanis spp., Trichoderma spp. and an unknown basidiomycete.
Genetic transformation, electrophoretic karyotyping and isolation of insertional mutants in the tree pathogenic fungus Neonectria galligena
Authors: Tanguay P. , Coupal J. , Bernier L.
Published: December 2004

The ascomycete tree pathogen Neonectria galligena was transformed using several plasmid vectors containing the bacterial hygromycin phosphotransferase gene (hph) conferring resistance to hygromycin B. Protoplasts were produced by digesting germinating conidia in a Novozym 234 solution osmotically stabilized with NaCl. The protoplasts regenerated at ca. 70% on complete medium supplemented with either sucrose or NaCl. Up to 115 hygromycin B‐resistant transformants per microgram of transforming DNA per 108 protoplasts were obtained. Southern analysis of genomic DNA from the transformants, separated by either standard agarose gel electrophoresis or by pulsed field gel electrophoresis, confirmed that hygromycin B resistance resulted from integration of the vector. Transformants expressing the green fluorescent protein were also obtained. A total of 209 transformants were then screened for aggressiveness and mycelial radial growth. Four mutants were identified including one mutant with reduced aggressiveness but normal growth in vitro, two mutants with reduced growth in vitro but normal aggressiveness, and one mutant in which both aggressiveness and in vitro growth were reduced.
Sequential activation of Notch family receptors during mouse spermatogenesis
Authors: Mori Shintaro , Kadokawa Yuzo , Hoshinaga Kiyotaka , Marunouchi Tohru
Published: December 2004

The expression pattern of Notch family receptors during mouse spermatogenesis was examined by immunohistochemistry. The entire cytoplasm of spermatogonia, spermatocytes and spermatids showed staining with antibodies against extracellular domains of Notch1, 2 and 4. In contrast, the nuclei of spermatogonia showed staining with an antibody against the intracellular domain of Notch3, and the nuclei of spermatocytes and spermatids showed staining with antibodies against the intracellular domains of Notch1 and 4. During regeneration of spermatogonia in busulfan‐treated mice, the nuclei of all proliferating cells showed staining for the intracellular domain of Notch3. Western blot analysis showed that the molecular weights of the intracellular domains of Notch1 and 3 localizing in the nuclear fraction were smaller than those in the cytoplasmic fraction. This was consistent with the theory that the intracellular domain of Notch was cleaved in the cytoplasm and translocated to the nucleus. These results suggest that different Notch signals are sequentially activated during mouse spermatogenesis and control the proliferation and differentiation of spermatogenic stem cells.
Ocular tissue interactions during the development of human fetal neuroretina grafted into nude mice
Authors: Angioi Karine , Hatier Renée , Duprez Adrien
Published: December 2004

To determine the roles of different ocular tissues in the development of the human fetal neuroretina, a study ethically and technically impossible in human subjects, human embryonic and fetal retinas were heterotopically implanted into nude mice. Ninety‐five eyeballs were obtained from legally aborted 6‐ to 7‐week‐old embryos or 8‐ to 10‐week‐old fetuses. Ten isolated neuroretinas with vitreous but without pigment epithelium, 20 half‐eyeballs and 70 intact eyeballs, of which 12 had a thick layer of periocular tissue, were microsurgically grafted. Five intact eyeballs were used for reference. Over a period of 1–245 days, all of the grafts were removed for light and electron microscopy observations. All of the isolated neuroretinas had disappeared by the second day after transplantation. Grafts of the posterior section of the eyeball contained only some clusters of pigment epithelium, occasionally covered with undifferentiated neuroretinal cells. Grafts of the retrolental section of the eyeball contained small areas of dysplasic neuroretina with folds and rosettes. Grafts of the 70 intact eyeballs were successful, but only 26 showed normal histological organization of the choriocapillaris, the retinal pigment epithelium and the neuroretina in the posterior part of the posterior chamber. Photoreceptor differentiation was evident in these retinas after approximately 80 days of transplantation and was complete after 166 days. Their anterior part was always dysplasic, with occasional ciliary differentiation. Twenty‐three grafted eyeballs had a dysplasic neuroretina with folds, rosettes and necrotized areas. Twenty‐one were atrophic, 12 of which were the eyeballs grafted with periocular tissue. These results demonstrate the role of the fetal mesenchyme and pigment epithelium in the rapid revascularization, and subsequent survival and tissue organization, of the neuroretina. The stratified development of the neuroretina required a thin mesenchymal environment for revascularization of the graft by human vasculogenesis or neoangiogenesis and a normal retinal pigment epithelium for normal neuroretinal differentiation. When these conditions were not satisfied, the neuroretina disappeared or was dysplasic, partly necrotized or atrophic. This model might prove useful for a number of therapeutic or clinical studies.
Endoscopic ultrasound‐guided fine needle aspiration biopsy in esophageal and mediastinal diseases: Clinical indications and results
Authors: ARIMA MIWAKO , TADA MASAHIRO
Published: December 2004

Background:  Endoscopic ultrasound‐guided fine needle aspiration biopsy (EUS‐FNAB) was developed to attain endosonographical images in real time in endoscopic biopsy, just like in percutaneous biopsy with ultrasonic or computer‐tomographical images. The results of EUS‐FNAB in esophageal and mediastinal diseases were evaluated and clinical indications of this technique were investigated.

Methods:  The study was performed in 58 patients, consisting of 30 with esophageal or mediastinal tumors and 28 requiring mediastinal lymph node examination. The instruments were linear array EUS transducer PEF‐703FA (Toshiba‐Fujinon, Tokyo, Japan) and 21G Endosonopsy (Hakko Shoji, Tokyo, Japan). The aspirated material was recovered on a filter paper and was formalin‐fixed to be examined histopathologically.

Results:  The tumors measured 6–60 mm (mean 29 mm). Collection of tissue was successful in 95% of the patients, and diagnostic accuracy was 95%. The biopsy specimen was satisfactory to establish histological diagnosis in every case of 27 patients with malignant diseases. No complication was experienced.

Conclusion:  The EUS‐FNAB is indicated in cases where the technique of EUS‐guided puncture is required, or is considered optimum in view of the safety and in cases where histological diagnosis is critical for the decision of treatment program. In many cases of mediastinal diseases, not even a detection of lesion is feasible without this technique, let alone a collection of tissue. The EUS‐FNAB is thus performed as a first choice of examination to obtain biopsy specimen in such cases. Endoscsopic ultrasound‐guided lymph node puncture is applied in order to assess the appropriateness of endoscopic mucosal resection (EMR) in patients with esophageal cancer, to follow up the patients after EMR and chemoradiotherapy (CRT), and to evaluate the efficacy of neoadjuvant CRT.
Endoscopic band ligation for postpolypectomy gastric bleeding
Authors: TOMIYAMA RYOSAKU , KINJO FUKUNORI , HOKAMA AKIRA , MIYAGI TSUYOSHI , NAKASONE HIROKI , KINJO NAGISA , SAKUGAWA HIROSHI , SAITO ATSUSHI
Published: December 2004

We report a case of a patient in whom endoscopic band ligation was achieved for postpolypectomy gastric bleeding. A 76‐year‐old man visited our hospital because of anemia. Endoscopy revealed a gastric polyp, approximately 12 mm in diameter, on the lesser curvature in the distal gastric body. The polyp was considered to be the source of chronic anemia and was therefore removed by using standard careful snare‐cautery polypectomy technique. Four days later, follow‐up endoscopy was performed to evaluate the postpolypectomy site, and an active bleeding postpolypectomy ulcer was identified. Initial attempts to achieve hemostasis with ethanol injection were unsuccessful. Immediate hemostasis was obtained with a subsequent endoscopic band ligation device. There has been no recurrent bleeding. Endoscopic band ligation might be a good treatment modality for the treatment of a postpolypectomy gastric bleeding lesion.
Application of percutaneous endoscopic gastrostojejunostomy in a case of amyotrophic lateral sclerosis associated with the superior mesenteric artery syndrome
Authors: KIKUI SHOJI , YOKOSE YOSHIHIKO , SUGIMOTO YASUSHI , INOUE HIROYA , KIN TESSEKI , SAKURAI RITSURO
Published: December 2004

A 64‐year‐old woman who was diagnosed as suffering from amyotrophic lateral sclerosis (ALS) of bulbar type was admitted to our hospital for long‐term care. After admission, she underwent percutaneus endoscopic gastrostomy (PEG) and enteral feeding was initiated. However, the PEG alimentation was disrupted by aspiration pneumonia owing to the superior mesenteric artery syndrome (SMAS), diagnosed by gross anatomy and endoscopic studies. Conservative treatment for SMAS was not successful and sepsis developed. After recovery from this, an operation was recommended, but it was rejected by her and her family members. We therefore selected the method of placing a thin jejunostomy tube through the PEG, called percutaneous endoscopic gastrojejunostomy (PEGJ) and pulling it endoscopically into the proximal jejunum, thereby allowing delivery of nutrients. Thereafter, she was well and showed gradual improvement of nutritional parameters such as serum albumin and total cholesterol, as well as the lymphocyte subset. It is concluded that PEGJ is effective for long‐term enteral nutrition in ALS patients complicated with SMAS.
Technique of laparoscopic cholecystectomy combined with percutaneous papillary balloon dilatation under general anesthesia for cholecystocholedocholithiasis
Authors: CHIKAMORI FUMIO , KUNIYOSHI NOBUTOSHI , KAWASHIMA TAKAHIKO , Takase Yasuhiro
Published: December 2004

Since the introduction of laparoscopic cholecystectomy (LC), the treatment of cholecystocholedocholithiasis has become a controversial issue among surgeons and endoscopists all over the world. We evaluated the effectiveness of LC combined with percutaneous papillary balloon dilatation (PPBD) under general anesthesia in the treatment of cholecystocholedocholithiasis in 22 patients. All stones in the bile duct were successfully evacuated into the duodenum in all patients. The PPBD was feasible in all patients under general anesthesia. The mean postoperative stay was 9 days. The overall length hospital stay and the duration of PTBD were 19 ± 7 days and 16 ± 8 days, respectively. There were no deaths nor major complications, although a transient hyperamylasemia was found in 10 patients (45%). Cholecystocholedocholithiasis was able to be treated by means of LC combined with PPBD under general anesthesia without laparotomy, sphincterotomy or choledochotomy. This technique can be a choice for the treatment that enables a patient to avoid any discomfort arisen as a result of papillary dilatation.
Cloning and functional analysis of alkB genes in Alcanivorax borkumensis SK2
Authors: Hara Akihiro , Baik Sang‐ho , Syutsubo Kazuaki , Misawa Norihiko , Smits Theo H. M. , Van Beilen Jan B. , Harayama Shigeaki
Published: December 2004

Alcanivorax is an alkane‐degrading marine bacterium which propagates and becomes predominant in crude‐oil‐containing seawater when nitrogen and phosphorus nutrients are supplemented. To identify the genes responsible for alkane degradation in this organism, two putative genes for alkane hydroxylases were cloned from Alcanivorax borkumensis SK2. They were named alkB1 and alkB2. These genes were subsequently disrupted in A. borkumensis SK2, and the growth phenotypes of the disruptants were examined. The results indicate that the alkB1 gene is responsible for the degradation of short‐chain n‐alkanes. A double mutant defective in both alkB1 and alkB2 was still able to grow on medium‐chain n‐alkanes, indicating that genes other than alkB1 and alkB2 are also involved in n‐alkane hydroxylation by A. borkumensis SK2.