bluuebunny/crossref_metadata_2025 huggingface dataset:
In vitro effects of flunisolide on MMP‐9, TIMP‐1, fibronectin, TGF‐β1 release and apoptosis in sputum cells freshly isolated from mild to moderate asthmatics
Authors: Profita M. , Gagliardo R. , Di Giorgi R. , Bruno A. , Riccobono L. , Bonanno A. , Bousquet J. , Vignola A. M.
Published: August 2004

Background:  Corticosteroids play an important role in inflammation and remodelling of airways and are considered an important therapeutic target in asthma. Inflammation in asthma is characterized by a dysregulation of eosinophil apoptosis and of markers of airways remodelling. We evaluated the ability of flunisolide to inhibit in vitro the release of metalloproteinases‐9 (MMP‐9), tissue inhibitor metalloproteinases‐1 (TIMP‐1), transforming growth factor (TGF‐β) and fibronectin by sputum cells (SC) as well as to induce sputum eosinophil apoptosis.

Methods:  The SC, isolated from induced sputum samples of 12 mild‐to‐moderate asthmatics, were cultured for 24 h in the presence or absence of flunisolide (1, 10 and 100 μM). The release of mediators was assessed by enzyme‐linked immunosorbent assay (ELISA) whereas apoptosis was studied by TUNEL technique.

Results:  Flunisolide (10 μM) significantly reduced MMP‐9 and TIMP‐1 (P = 0.0011 and P < 0.0001 respectively) and increased MMP‐9/TIMP‐1 molar ratio (P = 0.004). In addition, flunisolide decreased TGF‐β and fibronectin release by SC (P = 0.006; and P < 0.0001 respectively) and increased eosinophil apoptosis (P < 0.001).

Conclusions:  These results demonstrate that flunisolide may play an important role in the inhibition of airway inflammation and remodelling, by promoting the resolution of eosinophilic inflammation and by inhibiting the release of MMP‐9, TIMP‐1, TGF‐β and fibronectin.
Mouse models of global airway allergy: what have we learned and what should we do next?
Authors: Hellings P. W. , Ceuppens J. L.
Published: August 2004

Recent epidemiological and clinical data indicate that allergic rhinitis and asthma coexist and should be considered as one airway allergy syndrome. In spite of the importance of this new concept of global airway allergy, it has not fundamentally changed our daily diagnostic and therapeutic strategies so far because of the lack of essential clues to understand the correlation between allergic inflammation in upper and lower airways. Because of the resemblance of experimentally induced allergic airway inflammation in mice to inflamed airways of allergic patients, mouse models can enhance our insight into mechanisms underlying the global airway allergy syndrome. We here review data generated in mice that are relevant for understanding the development of airway allergy and provide new options for research on the so‐called ‘united airway disease’.
Aeroallergen sensitization in pediatric allergic rhinitis in Singapore: Is air‐conditioning a factor in the tropics?
Authors: Kidon Mona Iancovici , See Yvonne , Goh Anne , Chay Oh M. , Balakrishnan Abhilash
Published: August 2004

Singapore is a unique blend of a tropical environment with a high standard of hygiene and public health care. The objective was to define the prevalence, clinical characteristics, and environmental risk factors of specific aeroallergen sensitization in pediatric allergic rhinitis patients in this unique environment. The method adopted was a retrospective analysis of allergic rhinitis patients, undergoing aeroallergen skin prick testing (SPT), in the outpatient specialty clinic of the KK Children's hospital, from July 2001 to June 2002. A total of 202 patients were included, 161 (80%) males, 167 (83%) Chinese, age mean 7.6 yr (range 2–14 yr). The most prevalent clinical symptoms were: watery rhinorrhea 61%, blocked nose 61%, sneezing 52%, snoring 17%, and epistaxis 12%. SPT results were positive for house dust mites in 97% of children, pets (20%), molds (19%), pollens (15%), and kapok (10%). Mold sensitization was significantly more prevalent in households without air‐conditioning (aircon), 49% vs. 10% with aircon (odds ratio 9.4, 95% CI 3.8–22.9). Polysensitization (sensitization to three or more allergens) was similarly more prevalent in households without aircon, 51% vs. 14% with aircon (odds ratio 6.4, 95% CI 2.8–14.7). It was concluded that indoor aeroallergen sensitization is the major associated factor with clinical allergic rhinitis in children in Singapore. Patients living in households without air‐conditioning are at increased risk of mold sensitization and polysensitization.
Management of childhood asthma: Physicians’ perspective in Turkey
Authors: Civelek E. , Sekerel B. E.
Published: August 2004

Many attempts have been made to decrease asthma‐related mortality and morbidity, including preparation and distribution of guidelines. However, several studies from different regions of the world have reported low asthma control among children as well as adults. A questionnaire‐based study was performed involving a number of Turkish physicians taking care of pediatric asthmatics, and aimed to document asthma‐related knowledge. Of 219 physicians, 126 (57%) completed the questionnaire. The majority were pediatricians (92%), with an average of 253 ± 14.1 outpatients per week, and an asthmatic ratio of 15.8%. According to the self‐reported knowledge, management of acute asthma received the highest (3.42 ± 0.05) and chronic asthma the lowest scores (3.18 ± 0.06). The study group further credited inhaled and oral steroids similarly for the treatment of exacerbations and reported infrequent usage of spirometry and oxygen saturation than physical finding to estimate attack severity. Although with regard to most subjects, physicians’ opinions appeared to be in agreement with the guidelines, there were certain differences implying need for future educational training. Chronic management, reported as the least understood part of the guidelines, suggests requirement of new models for estimating chronic disease severity.
Invasion of broad‐leaf tree species into a larch plantation: seasonal light environment, photosynthesis and nitrogen allocation
Authors: Kitaoka Satoshi , Koike Takayoshi
Published: August 2004

We have studied how tree seedlings with differing leaf phenological traits change their nitrogen allocation in seasonally changing light environments. Specifically, we have investigated seasonal changes in maximum leaf photosynthetic rates in situ, and changes in nitrogen, chlorophyll and ribulose‐1,5‐bisphosphate carboxylase oxygenase (Rubisco) content. We have also estimated nitrogen allocation characteristics from leaf nitrogen to photosynthetic proteins, light‐harvesting protein and other proteins in four successional tree species growing in a larch plantation. The gap phase species, Magnolia hyporeuca Siebold et Zucc., displays higher photosynthetic rates to compensate for the short leafy period and allocates more nitrogen to Rubisco. In contrast, tree species that grow at the forest floor, Prunus ssiori F. Schmidt and Carpinus cordata Blume, use the leafless period of the canopy trees to change their allocation of nitrogen from Rubisco to chlorophyll in line with canopy closure. Quercus mongolica Fischer ex Ledeb. var. crispula (Blume) Ohashi is a mid‐successional tree species, and maintains a high photosynthetic rate in September because it allocates more nitrogen to Rubisco. We conclude that phenological differences in tree seedlings are clearly reflected in the nitrogen allocation pattern at gaps and at the forest floor in larch plantations.
Expression of the maize GALACTINOL SYNTHASE gene family: (II) Kernel abscission, environmental stress and myo‐inositol influences accumulation of transcript in developing seeds and callus cells
Authors: Zhao Tian‐Yong , Martin David , Meeley Robert B. , Downie Bruce
Published: August 2004

Accumulation of transcript from three maize (Zea mays L. inbred line B73) GALACTINOL SYNTHASE (ZmGOLS) genes in abscised developing seeds and callus before and after stress was examined. Previous work showed that transcript from ZmGOLS3 predominates during seed development, whereas ZmGOLS2 transcript accumulates when triggered by certain stresses following imbibition. Abscission stimulated accumulation of ZmGOLS transcript in immature kernels. This accumulation was enhanced by increasing durations of dehydration stress. In callus tissue, neither ZmGOLS1 nor ZmGOLS3 was detected regardless of media composition or the nature or duration of the applied stress. Accumulation of ZmGOLS2 transcript in heat‐ or dehydration‐stressed callus was induced by exogenously supplied myo‐inositol in low amounts but not by sucrose. However, abundance of transcript was a poor indicator of activity of GOLS or amounts of galactinol/raffinose regardless of type of tissue, composition of media, or imposition of stress. Neither were amounts of galactinol/raffinose tightly associated with activity of GOLS nor amounts of myo‐inositol, despite the ability of exogenously supplied raffinose to maintain the growth of dehydration‐stressed callus. The abundance of ZmGOLS transcript did not increase in response to exogenously supplied abscisic acid.
Accumulation pattern and identification of seed storage proteins in zygotic embryos of Pinus strobus and in somatic embryos from different maturation treatments
Authors: Klimaszewska Krystyna , Morency Frédéric , Jones‐Overton Catherine , Cooke Janice
Published: August 2004

Somatic embryogenesis (SE) of Pinus strobus L. has been greatly improved over the last few years with respect to both the initiation frequencies from a number of seed families and production of mature somatic embryos that readily convert to plants. However, there are no data on biochemical characterization of somatic embryos in relation to zygotic embryos of eastern white pine and on the optimal duration of the maturation stage. It is believed that somatic embryos closely resembling zygotic embryos not only morphologically but biochemically would display more vigorous growth. Hence, in this study the accumulation pattern of the most abundant seed storage proteins in zygotic and somatic embryos were characterized by sodium dodecyl sulphate‐polyacrylamide gel electrophoresis (SDS‐PAGE) and identified by amino acid sequencing and tandem mass spectrometry (MS/MS). This showed that somatic embryos accumulated storage proteins in a similar manner to zygotic embryos and that the most abundant were the buffer‐insoluble 11S‐ globulins MW 59.6 kDa, which dissociated under reduced conditions to 38.2–40.0 and 22.5–23.5 kDa range polypeptides, and buffer‐soluble 7S vicilin‐like proteins MW 46.0–49.0 kDa, which did not separate under reduced conditions. Other relatively abundant soluble proteins were in the ranges of 25–27 and 27–29 kDa. The only group of proteins that showed different migration profiles in the presence of β‐mercaptoethanol (ME) were the low molecular mass proteins of 14.6–16.5 kDa. Somatic embryos that matured for 9 weeks on medium with 6% sucrose accumulated more storage proteins than those matured on medium with 3% sucrose and the extension of the maturation period to 12 weeks resulted in significant reduction of the storage proteins on both media. As expected, somatic embryos matured on medium with 6% sucrose had lower water potential (Ψ) than those from medium with 3% sucrose. Nonetheless, the somatic embryos matured under the best of tested conditions (6% sucrose for 9 weeks) had slightly higher water content; 1.35 ± 0.28 g H2O g−1 DM (mean ± sd) than the mature non‐dried zygotic embryos; (1.16 ± 0.09 g H2O g−1 DM), and accumulated less storage proteins, whose amounts were either similar to (7S‐vicilins) or below (11S‐globulins) those found in the immature zygotic embryos collected 2 weeks prior to the usual cone collection. The implications of these results for further research and development of viable artificial seed is discussed.
Iron deficiency induces changes in metabolism of citrate in lateral roots and cluster roots of Lupinus albus
Authors: McCluskey Joyce , Herdman Lindsey , Skene Keith Ronald
Published: August 2004

This paper describes the first measurement of enzyme activities in cluster roots under –Fe stress, at different stages of cluster root development and function. In Lupinus albus L., Cluster roots are produced both under iron‐ and phosphorus‐deficient conditions. In both cases the structure is similar, but the level of exudation is much greater in iron‐deficient plants. Much work has been done on the enzyme kinetics of P‐deficient cluster roots, but none on enzyme activities of Fe‐deficient cluster roots. The enzymes investigated were citrate synthase (EC 4.1.3.7), aconitase (EC 4.2.1.3), isocitrate dehydrogenase [IDH(NAD) (EC 1.1.1.41) and IDH (NADP) (EC 1.1.1.42)] and lactate dehydrogenase (LDH) (EC 1.1.1.27). In cluster roots, citrate synthase activity was initially lower than in lateral roots but, after 5 days, recovered to the lateral root level. Cluster root aconitase levels initially increased, but fell sharply on day 3, and no activity was detected after day 5. IDH (NAD) levels were much lower in cluster roots than in laterals, dropping to a low on day 3, and then rising throughout development. IDH (NADP) levels were always higher in cluster roots than in lateral roots, increasing throughout development. LDH levels in cluster roots fell throughout development. Internal tissue concentrations of citrate were markedly higher in –Fe laterals than in +Fe lateral roots and in cluster roots. Cluster root levels of citrate increased dramatically after day 3. Results are discussed within the context of previous work on enzyme kinetics under –P, and the importance of a block in aconitase activity is highlighted.
Akt‐dependent phosphorylation negatively regulates the transcriptional activity of dHAND by inhibiting the DNA binding activity
Authors: Murakami Masao , Kataoka Keiichiro , Fukuhara Shigetomo , Nakagawa Osamu , Kurihara Hiroki
Published: August 2004

HAND2/dHAND is a basic helix‐loop‐helix transcription factor expressed in the heart and neural crest derivatives during embryogenesis. Although dHAND is essential for branchial arch, cardiovascular and limb development, its target genes have not been identified. The regulatory mechanisms of dHAND function also remain relatively unknown. Here we report that Akt/PKB, a serine/threonine protein kinase involved in cell survival, growth and differentiation, phosphorylates dHAND and inhibits dHAND‐mediated transcription. AU5‐dHAND expressed in 293T cells became phosphorylated, possibly at its Akt phosphorylation motif, in the absence of kinase inhibitors, whereas the phosphatidylinositol 3‐kinase inhibitor wortmannin and the Akt inhibitor NL‐71‐101, but not the p70 S6 kinase inhibitor rapamycin, significantly reduced dHAND phosphorylation. Coexpression of HA‐Akt augmented dHAND phosphorylation at multiple serine and threonine residues mainly located in the bHLH domain and, as a result, decreased the transcriptional activity of dHAND. Consistently, alanine mutation mimicking the nonphosphorylation state abolished the inhibitory effect of Akt on dHAND, whereas aspartate mutation mimicking the phosphorylation state resulted in a loss of dHAND transcriptional activity. These changes in dHAND transcriptional activity were in parallel with changes in the DNA binding activity rather than in dimerization activity. These results suggest that Akt‐mediated signaling may regulate dHAND transcriptional activity through the modulation of its DNA binding activity during embryogenesis.
GlnK effects complex formation between NifA and NifL in Klebsiella pneumoniae
Authors: Stips Jessica , Thummer Robert , Neumann Melanie , Schmitz Ruth A.
Published: August 2004

In Klebsiella pneumoniae, the nif specific transcriptional activator NifA is inhibited by NifL in response to molecular oxygen and ammonium. Here, we demonstrate complex formation between NifL and NifA (approximately 1 : 1 ratio), when synthesized in the presence of oxygen and/or ammonium. Under simultaneous oxygen‐ and nitrogen‐limitation, significant but fewer NifL–NifA complexes (approximately 1%) were formed in the cytoplasm as a majority of NifL was sequestered to the cytoplasmic membrane. These findings indicate that inhibition of NifA in the presence of oxygen and/or ammonium occurs via direct NifL interaction and formation of those inhibitory NifL–NifA complexes appears to be directly and exclusively dependent on the localization of NifL in the cytoplasm. We further observed evidence that the nitrogen sensory protein GlnK forms a trimeric complex with NifL and NifA under nitrogen limitation. Binding of GlnK to NifL–NifA was specific; however the amount of GlnK within these complexes was small. Finally, two lines of evidence were obtained that under anaerobic conditions but in the presence of ammonium additional NtrC‐independent GlnK synthesis inhibited the formation of stable inhibitory NifL–NifA complexes. Thus, we propose that the NifL–NifA–GlnK complex reflects a transitional structure and hypothesize that under nitrogen‐limitation, GlnK interacts with the inhibitory NifL–NifA complex, resulting in its dissociation.
Val216 decides the substrate specificity of α‐glucosidase in Saccharomyces cerevisiae
Authors: Yamamoto Keizo , Nakayama Akifumi , Yamamoto Yuka , Tabata Shiro
Published: August 2004

Differences in the substrate specificity of α‐glucosidases should be due to the differences in the substrate binding and the catalytic domains of the enzymes. To elucidate such differences of enzymes hydrolyzing α‐1,4‐ and α‐1,6‐glucosidic linkages, two α‐glucosidases, maltase and isomaltase, from Saccharomyces cerevisiae were cloned and analyzed. The cloned yeast isomaltase and maltase consisted of 589 and 584 amino acid residues, respectively. There was 72.1% sequence identity with 165 amino acid alterations between the two α‐glucosidases. These two α‐glucosidase genes were subcloned into the pKP1500 expression vector and expressed in Escherichia coli. The purified α‐glucosidases showed the same substrate specificities as those of their parent native glucosidases. Chimeric enzymes constructed from isomaltase by exchanging with maltase fragments were characterized by their substrate specificities. When the consensus region II, which is one of the four regions conserved in family 13 (α‐amylase family), is replaced with the maltase type, the chimeric enzymes alter to hydrolyze maltose. Three amino acid residues in consensus region II were different in the two α‐glucosidases. Thus, we modified Val216, Gly217, and Ser218 of isomaltase to the maltase‐type amino acids by site‐directed mutagenesis. The Val216 mutant was altered to hydrolyze both maltose and isomaltose but neither the Gly217 nor the Ser218 mutant changed their substrate specificity, indicating that Val216 is an important residue discriminating the α‐1,4‐ and 1,6‐glucosidic linkages of substrates.
CENTRAL EFFECT OF HISTAMINE AND PERIPHERAL EFFECT OF HISTIDINE ON THE FORMALIN‐INDUCED PAIN RESPONSE IN MICE
Authors: Tamaddonfard Esmaeal , Rahimi Saead
Published: August 2004

1. The present study was designed to investigate the role of brain histamine in modulating pain transmission in mice.

2. In conscious mice implanted with an intracerebroventricular (i.c.v.) cannula, the effects of i.v.c. injections of normal saline (control) and low and high doses histamine (2 and 40 µg/mouse, respectively) were investigated on the duration of paw licking and biting induced by subcutaneous (s.c.) injection of formalin (20 µL; 5%) into the plantar surface of the left hindpaw.

3. To clarify the involvement of histidine in the pain response, the effects of intraperitoneal (i.p.) injections of low and high doses of histidine (50 and 1000 mg/kg, respectively) alone or before i.c.v. injection of histamine were also examined.

4. Intraplantar injection of formalin induced a biphasic pain response (first phase: 0–5 min after injection; second phase: 20–40 min after injection).

5. Histamine (2 µg/mouse, i.c.v.) had no effect on the first phase of the pain response, but suppressed the second phase. The higher dose of histamine (40 µg/mouse, i.c.v.) suppressed both phases of the pain response.

6. Histidine, at 50 mg/kg, i.p., had no effect on the pain response, but the higher dose (1000 mg/kg, i.p.) suppressed the both phases of the pain response.

7. Pretreatment with the low dose of histidine (50 mg/kg, i.p.) prior to administration of 2 µg/mouse, i.c.v., histamine did not change the antinociception induced by low‐dose histamine. However, pretreatment with the high dose of histidine (1000 mg/kg, i.p.) prior to 2 µg/mouse, i.c.v., histamine produced antinociception that resembled that seen following administration of the high dose of either histidine or histamine. Pretreatment with the low dose of histidine (50 mg/kg, i.p.) prior to administration of 40 µg/mouse, i.c.v., histamine has no effect on the pain response following high‐dose histamine. Pretreatment with 1000 mg/kg, i.p., histidine prior to administration of 40 µg/mouse, i.c.v., histamine strongly suppressed both phases of the formalin‐induced pain response, particularly the second phase.

8. The results of the present study indicate that: (i) activation of brain histamine produces antinociception in the mouse formalin test; (ii) peripheral loading with a high dose of histidine (1000 mg/kg, i.p.) alone exerts the same effect as that seen following 40 µg/mouse, i.c.v., histamine; and (iii) pretreatment with a high dose of histidine potentiates central histamine‐induced antinociception.
Thrust geometries in unconsolidated Quaternary sediments and evolution of the Eupchon Fault, southeast Korea
Authors: Kim Young‐Seog , Park Joon Young , Kim Jeong Hwan , Shin Hyeon Cho , Sanderson David J.
Published: July 2004

Abstract  The Korean peninsula is widely regarded as being located at the relatively stable eastern margin of the Asian continent. However, more than 10 Quaternary faults have recently been discovered in and reported from the southeastern part of the Korean Peninsula. One of these, the Eupchon Fault, was discovered during the construction of a primary school, and it is located close to a nuclear power plant. To understand the nature and characteristics of the Quaternary Eupchon Fault, we carried out two trench surveys near the discovery site. The fault system includes one main reverse fault (N20°E/40°SE) with approximately 4 m displacement, and a series of branch faults, cutting unconsolidated Quaternary sediments. Structures in the fault system include synthetic and antithetic faults, hanging‐wall anticlines, drag folds, back thrusts, pop‐up structures, flat‐ramp geometries and duplexes, which are very similar to those seen in thrust systems in consolidated rocks. In the upper part of the fault system, several tip damage zones are observed, indicating that the fault system propagates upward and terminates in the upper part of the section. Pebbles along the main fault plane show a preferred orientation of long axes, indicating the fault trace. The unconformity surface between the Quaternary deposits and the underlying Tertiary andesites or Cretaceous sedimentary rocks is displaced by this fault with a reverse movement sense. The stratigraphic relationship shows normal slip sense at the lower part of the section, indicating that the fault had a normal slip movement and was reversely reactivated during the Quaternary. The inferred length of the Quaternary thrust fault, based on the relationship between fault length and displacement, is 200–2000 m. The current maximum horizontal compressive stress direction in this area is generally east‐northeast–west‐southwest, which would be expected to produce oblique slip on the Eupchon Fault, with reverse and right‐lateral strike‐slip components.
Evidence for high‐Ca boninite magmatism from Paleogene primitive low‐K tholeiite, Mukoojima, Hahajima Island group, southern Bonin (Ogasawara) forearc, Japan
Authors: Maehara Kosuke , Maeda Jinichiro
Published: July 2004

Abstract  High‐Ca boninitic inclusions are found in primitive low‐K tholeiite from Mukoojima (Mukoo‐Jima), an islet in the Hahajima Island group, Bonin (Ogasawara) forearc, Japan. While Chichijima Island group, 50 km north of Hahajima Island group, is well known as a type locality of boninite, there has been no report of boninitic rocks from the Hahajima Island group. The high‐Ca boninitic inclusions are aphanitic and contain olivine, Ca‐rich clinopyroxene, plagioclase, chromian spinel, opaque minerals and dark brown glass. The mode of occurrence of the inclusions and host tholeiite under the microscope indicates mingling of these two magmas, suggesting intimate association in space and time of the boninite and primitive tholeiite magmas around the Hahajima Island group in Paleogene time. Primitive compositions and slightly different Sr and Nd isotopic ratios suggest that these two magmas are derived from two distinct mantle sources. These two mantle sources were present at the same time around the Hahajima Island group, southern Bonin forearc. The source of the high‐Ca boninite was higher in water content and/or shallower in depth compared to that of the primitive tholeiite.
K–Ar geochronology of the temporal change of eruptive style in the eastern Izu peninsula, central Japan
Authors: Ozawa Ayako , Tagami Takahiro , Sudo Masafumi
Published: July 2004

Abstract  A recent K–Ar study elucidated that eruptive style in the eastern Izu peninsula changed from polygenetic to monogenetic volcano at 0.3–0.2 Ma. To narrow down the time of change, we determined 10 K–Ar ages on Togasayama Andesite of Amagi volcano, the youngest polygenetic volcano in the area, and Togasayama Monogenetic Volcano, one of the oldest monogenetic volcanoes in the area, which overlies a part of the Togasayama Andesite. Dating results showed that the Togasayama Andesite effused at least from 0.34 to 0.20 Ma, whereas the Togasayama Monogenetic Volcano erupted at 0.26–0.29 Ma, suggesting that the northern part of the Togasayama Andesite effused after the eruption of the Togasayama Monogenetic Volcano. Considering previous data, it is therefore inferred that change of eruptive style in the eastern Izu area occurred during the period 0.29–0.20 Ma, with considerable overlap of both polygenetic and monogenetic volcanism.
C677T mutation in the MTHFR gene was not found in patients with frontoethmoidal encephalocele in East Java, Indonesia
Authors: Sadewa Ahmad Hamim , Sutomo Retno , Istiadjid Mohamad , Nishiyama Kaoru , Shirakawa Taku , Masafumi Matsuo  , Nishio Hisahide
Published: August 2004

Abstract
Background
: Frontoethmoidal encephalocele (FEE) is a neural tube defect (NTD) characterized by a congenital bone defect in the anterior cranium and herniation of the intracranial mass through the defect. The C677T mutation in the 5,10‐methylenetetrahydrofolate reductase gene (MTHFR) has been reported as a genetic risk factor for spina bifida. However, the role of the MTHFR in the pathogenesis of FEE remains to be clarified.

Methods
: A hospital‐based survey of FEE patients who were referred to the Department of Neurosurgery and Plastic Surgery, Malang General Hospital, East Java, Indonesia was conducted. Genetic screening of MTHFR substitutions in 13 patients and eight mothers from 11 affected families were performed using a combination of polymerase chain reaction (PCR), denaturing high‐performance liquid chromatography (DHPLC), and direct sequencing.

Results
: In total, 130 patients with FEE among 138 NTD patients (94.2%) were identified. The ratios of cranial encephalomeningocele to spinal meningocele (32 : 1) and of FEE to occipital encephalomeningocele (32 : 1) were higher than those in other populations. Five substitutions were detected in the MTHFR: C121T, C677T, C1060T, A1298C, and G1793A. No significant differences were found in the frequency of each nucleotide substitution between patients or mothers and controls. In addition, none of the subjects in this study were homozygous for T at nucleotide position 677.

Conclusion
: FEE is the most common form of NTD in East Java, Indonesia. Genetic analysis of 11 affected families suggests that the MTHFR gene is not associated with the development of FEE, although the number of FEE families analyzed in this study was very limited.
Comparative study of perceptions of work environment and moral sensitivity among Japanese and Norwegian nurses
Authors: Bégat Ingrid , Ikeda Noriko , Amemiya Takiko , Emiko Konishi , Iwasaki Akiko , Severinsson Elisabeth
Published: August 2004

Abstract The aim of the present study was to explore the relationship between work environment and moral sensitivity among Japanese (n = 138) and Norwegian nurses (n = 71), and to compare the results from a sociocultural perspective using a descriptive‐correlational design. Data were analyzed using descriptive and inferential statistics. The results point to a significant relationship between work environment and moral sensitivity for both groups of nurses. In comparison, the Japanese nurses were more focused on ‘patient centered oriented care’, reported ‘work engagement’, seeking ‘meaning in difficult caring situations’ and ‘following rules’. In addition, they ranked the factor ‘values in action of patient care’ as significant and ‘relation to superior and colleagues’ and ‘job stress and anxiety’ ranked significant to ‘moral conflicts’. The Norwegian nurses were more independent, which was correlated with moderate significance with ‘job stress and anxiety’. A significant correlation was found between ‘physical and mental symptoms’ and ‘moral conflicts’ among Norwegian nurses.
Smoking as a symbol of friendship: Qualitative study of smoking behavior and initiation of a group of male nurse students in Iran
Authors: Nikbakht Nasrabadi Alireza , Parsayekta Zohreh , Emami Azita
Published: August 2004

Abstract The study of smoking initiation and behavior of nurses and nurse students is important when considering their professional health promotion roles. In the present qualitative study, 26 nursing students of all grades from two nursing schools in the capital city of Iran were surveyed to explore their reasons for smoking. Through in‐depth and semistructured interviews, participants were encouraged to talk about their smoking behavior and explain initiating reasons freely. Majority of the participants began smoking before commencing nursing, while only a few initiated smoking after entering into nursing school. Reasons for smoking included belonging to a group and as a symbol of friendship and mutual trust. Indeed, peer group behaviors influenced most participates to commence smoking and social influences also emerged as a theme in this study.
Development of web‐based qualitative and quantitative data collection systems: Study on daily symptoms and coping strategies among Japanese rheumatoid arthritis patients
Authors: Kanzaki Hatsumi , Makimoto Kiyoko , Takemura Tadamasa , Ashida Nobuyuki
Published: August 2004

Abstract With the sharp rise in Internet access in recent years, the Internet is increasingly being used for research. We have developed two new systems using the Internet for both quantitative and qualitative data collection. One is a dynamic system for creating questionnaires and collecting responses and the other is an individual patient support follow‐up system. These systems do not depend on the capacity of computers or servers and enable researchers to interact with participants privately and asynchronously. We have used these systems to collect data on daily symptoms and journal entries from 12 community‐dwelling women with rheumatoid arthritis (RA) for a minimum of 1 month. The mean number of data submissions per month was 14.2 ± 7.8, with the majority recording entries every day, and some submitting several times a day. The combination of two types of data elucidated the changes in coping and coping strategies in conjunction with changes in symptoms, even in a single day, and the coexistence of positive and negative coping. Research participants benefited from web‐based symptom management and counseling resulting from 1 month of frequent interactions with the researcher. The use of the Internet for nursing research and interventions thus seems to show promising results.
Seed mass and seedling establishment after fire in Ku‐ring‐gai Chase National Park, Sydney, Australia
Authors: MOLES ANGELA T. , WESTOBY MARK
Published: August 2004

Abstract  Relationships between seed mass and several aspects of plant regeneration ecology were investigated in a post‐fire environment in Ku‐ring‐gai Chase National Park near Sydney, Australia. We found a significant positive relationship between seed mass and time to seedling emergence (P < 0.001) and a strong negative relationship between seed mass and time between emergence and production of the first true leaf (P < 0.001). Surprisingly, we found no relationship between seed mass and seedling establishment (P = 0.21). It seems most likely that this lack of relationship is a result of the many stochastic factors affecting seedling establishment during any given recruitment episode at any given site. A cause of mortality was assigned to 56% of the 781 seedlings that died during the present study. There was no relationship between cause of death and seed mass (P = 0.28). Of the seedlings for which the cause of death was known, 57% were killed by herbivory and 21% were killed by drought. Seedling–seedling competition affected only one species.
Seasonal variation in larval host plants and consequences for Heliconius erato (Lepidoptera: Nymphalidae) adult body size
Authors: RODRIGUES DANIELA , MOREIRA GILSON R. P.
Published: August 2004

Abstract  Adult body size, a key life history component, varies strongly within and between Heliconius erato phyllis (Lepidoptera: Nymphalidae) populations. In the present study, we determined whether seasonal variation in adult body size is temperature related and/or determined by seasonal changes of host plants (Passifloraceae) used by the larval stage. A population of H. erato phyllis located in a Eucalyptus plantation (Barba Negra Forest, Barra do Ribeiro County, Rio Grande do Sul State, Brazil) was sampled every 45 days from March 1997 to October 1998 to quantify seasonal variation in adult body size and use of larval host plants. In the laboratory, the effects of the following factors on adult body size were quantified: (i) host plant species (Passiflora misera or Passiflora suberosa); (ii) food quantity consumed by larvae (experimentally manipulated for each passion vine species); (iii) winter and summer temperatures (15 and 25°C, respectively); and (iv) the interaction between host plant species and temperature. Adults emerging during summer were larger than those emerging in other seasons. Female butterflies oviposited selectively on P. misera even when the dominant passion vine was P. suberosa. They only switched from using P. misera to P. suberosa during later autumn and winter, when P. misera vines were completely defoliated. The laboratory feeding trials with both passion vines showed a strong positive association between food quantity consumed by larvae and adult size. They also confirmed that adults are larger when their larvae are reared on P. misera than on P. suberosa. Temperature during larval development had no effect on H. erato phyllis adult size. Thus, seasonal variation of H. erato phyllis adult size in a given place is primarily determined by the availability and quality of host plant species used by the larval stage.
Microsite requirements, population structure and growth of the cushion plant Azorella compacta in the tropical Chilean Andes
Authors: KLEIER CATHERINE , RUNDEL PHILIP W.
Published: August 2004

Abstract  In the altiplano of northern Chile, we investigated the microsite habitats, adult population structure and growth rates over 14 months for the giant cushion plant Azorella compacta. The majority of plants (52.0%) were found growing downslope of large (>50 cm diameter) boulders and on north‐facing rocky slopes with a slope angle greater than 10° inclination (44.4%). Size was used as a surrogate for age. The youngest size class was most common; remaining size classes were roughly equal in frequency; but very large cushions were scarce. Growth rates were expressed as height, diameter and perimeter over a 14‐month period. Height increased an average of 1.40 cm, perimeter increased an average of 10.28 cm and diameters actually decreased 2.30 cm, although this was not a significant difference. Using perimeter averages based on mean plant size, we estimated a radial growth rate of 1.70 cm per 14 months. Trenches 4 cm wide were cut over the length of five plants, and these trenches were nearly repaired after 1 year, indicating that growth is overall and not just on the perimeter of the plant. These findings indicate that although Azorella compacta populations may be recovering from past exploitation, future monitoring is still necessary to ensure viable populations.
Germinable soil seedbanks of central Queensland rangelands invaded by the exotic weed Parthenium hysterophorus L.
Authors: NAVIE SHELDON C. , PANETTA F. DANE , McFADYEN RACHEL E. , ADKINS STEVE W.
Published: August 2004

The germinable soil seedbank was determined at two sites in central Queensland on four separate occasions between February 1995 and October 1996. These sites were infested with parthenium weed (Parthenium hysterophorus L.), a serious invasive exotic weed. During this period, the seedbank varied between 3282 and 5094 seeds m−2 at the Clermont site, and between 20 599 and 44 639 seeds m−2 at the Moolayember Creek site. Parthenium hysterophorus exhibited a very abundant and persistent seedbank, accounting for 47–73% of the seedbank at Clermont and 65–87% of the seedbank at Moolayember Creek. The species richness and species diversity of the seedbank, and the seed abundance of many species, was lower at Moolayember Creek during spring (the time of year when the most dense infestations of the weed originate). Parthenium hysterophorus seedlings also emerged more rapidly from the soil samples than did those of all other species. Hence, it seems that various aspects of the weed's seed ecology, including abundance and the persistence of its seedbank and the rapid emergence of its seedlings, are major factors contributing to its aggressiveness in semiarid rangeland communities in central Queensland. The domination by P. hysterophorus of the seedbanks of these sites suggests that the weed is having a substantial negative impact on the ecology of these plant communities. The diversity of these seedbanks was found to be lower in comparison with that observed in other grassland communities that were not dominated by an invasive weed species. Hence, the prolonged presence of P. hysterophorus may have substantially reduced the diversity of these seedbanks, thereby reducing the ability of some of the native species to regenerate in the future.
Identification of 66 kDa phosphoprotein associated with motility initiation of hamster spermatozoa
Authors: FUJINOKI MASAKATSU , KAWAMURA TAKESHI , TODA TOSHIFUSA , ISHIMODA‐TAKAGI TADASHI , OHTAKE HIDEKI , SHIMIZU NOBUYOSHI , OKUNO MAKOTO
Published: August 2004

Background and Aims:  Sperm motility is regulated by protein phosphorylation. The 66 kDa protein obtained from hamster sperm flagella was phosphorylated at serine residues associated with the motility initiation. In order to understand the regulatory mechanism of sperm motility, the 66 kDa protein was identified in the present study.

Methods:  The 66 kDa protein was purified by 2‐D gel electrophoresis and identified by matrix‐assisted laser desorption ionization mass spectrometry, liquid chromatography‐tandem mass spectrometry and peptide sequencer.

Results:  The 66 kDa protein was tubulin β chain.

Conclusion:  The 66 kDa protein is one of the tubulin β chain isoforms and phosphorylated in relation to the motility initiation. (Reprod Med Biol 2004; 3: 133–139)
Histological analysis of the ‘critical point’ in follicular development in mice
Authors: KAGABU SATOSI , UMEZU MOTOAKI
Published: August 2004

Aim:  To histologically examine ovarian follicular development in cycling mice.

Methods:   Mice were observed at 13:00 h at each stage of the estrous cycle. The ovaries were cut into complete serial sections. All sections were observed, and the size of each follicle was measured by using a micrometer. Follicles with advanced atresia were excluded and non‐atretic healthy follicles were differentiated from atretic follicles.

Results:  The measurement of the number of follicles in each stage of the estrous cycle, with reference to their size, showed that in each mouse, the number of small healthy follicles (100–249 µm in average diameter) was approximately 100 for all stages and the number of medium‐sized healthy follicles (250–349 µm) was close to 20 in only the metestrus and diestrus stages. In contrast, large healthy follicles (≥350 µm) showed marked changes throughout the estrous stage. Many healthy large follicles were observed in the proestrus stage, but they disappeared in the estrus stage, which suggests that they have ovulated. This was supported by observations of oocytes resuming meiosis in large healthy follicles.

Conclusion:  As follicular atresia was frequently observed in follicles of 250–399 µm diameter, this size range may be a ‘critical point’ for atresia. The results suggest that there is a ‘critical point’ in follicular development, and that only follicles that pass this point will ovulate, while those that do not will become atretic follicles. (Reprod Medical Biol 2004; 3: 141–145)
Epigenetic alteration by the chemical substances, food and environmental factors
Authors: FUKATA HIDEKI , MORI CHISATO
Published: August 2004

Epigenetic alteration is one of the most important mechanisms for gene regulation; however, it is not changes in gene function with DNA sequence changes. Recently, epigenetics were studied in the wide ranging fields of research. In the present review, we introduce recent studies on epigenetic alteration, especially DNA methylation, by chemical exposure, food intake and environmental factors. In addition, we introduced our results on alteration of DNA methylation by transient exposure of neonatal mice to diethylstilbestrol. As these data suggest that chemical exposure, food intake and environmental factors are responsible for epigenetic alteration, we insist the necessity of the new risk assessment focusing on epigenetic alteration. (Reprod Med Biol 2004; 3: 115–121)
Effects of orlistat on obesity‐related diseases – a six‐month randomized trial
Authors: Guy‐Grand B. , Drouin P. , Eschwège E. , Gin H. , Joubert J‐M. , Valensi P.
Published: August 2004

Aim:  To assess the effect of orlistat on body weight and concomitant diseases in patients with body mass index (BMI) of > 28 kg/m2 and poorly controlled type 2 diabetes, hypertension or hypercholesterolaemia.

Methods:  This trial was a six‐month, randomized, double‐blind, placebo‐controlled study of orlistat 120 mg three times daily plus a mildly reduced‐calorie diet. 1004 obese patients (BMI 28–40 kg/m2) were included by 253 private endocrinologists and received orlistat (n = 499) or placebo (n = 505). Patients were stratified by concomitant disorder (type 2 diabetes, n = 193; hypertension, n = 614; hypercholesterolaemia, n = 197). Body weight, anthropometry, lipid and glycaemic control parameters and blood pressure.

Results:  After six months, orlistat produced a significantly greater weight loss than placebo in type 2 diabetes (−4.2% vs. −1.4%), hypertension (−6.2% vs. −1.9%) and hypercholesterolaemia (−5.5% vs. −2.3%) groups (p < 0.0001 for all). There was a greater decrease in HbA1c in the type 2 diabetes group (−0.54 vs. −0.18%; p = 0.002) and low‐density lipoprotein (LDL)‐cholesterol in the hypercholesterolaemia group (−11.7% vs. −4.5%; p = 0.004) with orlistat vs. placebo. Early weight loss (≥ 5% at 12 weeks) was associated with the highest weight loss in each group, and the highest decreases in HbA1c, LDL‐cholesterol and diastolic blood pressure in patients with type 2 diabetes, hypercholesterolaemia and hypertension, respectively, at six months. The incidence of adverse events was similar for orlistat and placebo, except for certain generally well‐tolerated gastrointestinal events that were more common with orlistat.

Conclusion:  Orlistat plus a mildly reduced‐calorie diet produced clinically meaningful weight loss and improvements in risk factors in overweight and obese patients with poorly controlled type 2 diabetes, hypertension or hypercholesterolaemia.
Holmium:YAG laser vaporization of recurrent papillary tumours of the bladder under local anaesthesia
Authors: Jønler Morten , Lund Lars , Bisballe Søren
Published: August 2004

OBJECTIVES

To evaluate holmium:YAG laser vaporization of papillary tumours of the bladder, focusing on surgical technique, patients’ satisfaction, complications and cost‐effectiveness when using the technique under local anaesthesia as an outpatient procedure.

PATIENTS AND METHODS

In all, 52 consecutive patients with recurrent papillary tumours of the bladder were scheduled for holmium:YAG laser vaporization under local anaesthesia using a flexible cystoscope. The number of papillomas and total operative duration was recorded. Patients and surgeons were asked to complete a questionnaire about the procedure.

RESULTS

In all, 197 papillomas were successfully vaporized in 88 operations, with a median operative duration of 15 min (5 min per papilloma) and no patient needed treatment under general anaesthesia. Most patients (86%) had no pain (as reported during standard cystoscopy) and none of the procedures was stopped because of pain. All patients would undergo the treatment again, compared with a standard transurethral resection of bladder tumour. The five surgeons rated the procedure as easy in most patients (78%) and difficult in a few (6%). The total cost for the outpatient procedure was less than that for standard treatment.

CONCLUSION

This study clearly indicates that holmium:YAG laser vaporization of superficial bladder tumours is feasible, easy and fast, with a high degree of patient satisfaction, and it seems to be an attractive alternative to standard treatment. The procedure has some clear positive socio‐economic perspectives in both the short‐ and long‐term.
Early and large‐dose intravesical instillation of epirubicin to prevent superficial bladder carcinoma recurrence after transurethral resection
Authors: Mitsumori Kenji , Tsuchiya Norihiko , Habuchi Tomonori , Li Zhenhua , Akao Toshiya , Ohyama Chikara , Sato Kazunari , Kato Tetsuro
Published: August 2004

OBJECTIVE

To prospectively compare the prevention of tumour recurrence by four intravesical adjuvant administration protocols, and thus elucidate the efficacy of early and high total dose instillations of epirubicin to prevent superficial bladder tumour recurrence after transurethral resection of bladder tumour (TURBT).

PATIENTS AND METHODS

In all, 69 patients with Ta/T1 bladder cancer were randomly assigned to four intravesical administration protocols: A, delayed instillation (first instillation 7 days after TURBT) and low‐dose (30 mg once every 2 weeks, six times): B, early instillation (three instillations before 7 days after TURBT) and low‐dose; C, delayed and high‐dose (30 mg once weekly 12 times) instillation; D, early and high‐dose. The influence of the instillation protocols and tumour characteristics on the probability of recurrence‐free survival was examined using Kaplan‐Meier analysis and a Cox regression hazard model.

RESULTS

The early‐instillation and high‐dose groups had relatively lower recurrence rates after 6 months (A, 30%; B, 25%; C, one of 12; and D, none) and 1 year (50%, 35%, four of nine and one of eight, respectively). Patients who received 360 mg epirubicin (C and D) had a significantly better recurrence‐free survival than those receiving 180 mg (A and B; P = 0.012). Preoperative urine cytology and tumour multiplicity were significantly associated with recurrence. However, multivariate analysis of the risk of recurrence using a Cox proportional hazard model showed that urine cytology (hazard ratio 3.11, 95% confidence interval 1.08–8.94, P = 0.04) and total dose (0.32, 0.11–0.92, P = 0.03) were independent prognostic factors for recurrence.

CONCLUSION

Patients who received a high‐dose epirubicin instillation had a significantly lower recurrence rate but the benefit of early instillation was not confirmed, as the study group was too small.