bluuebunny/crossref_metadata_2025 huggingface dataset:
Protein complexes of the plant plasma membrane resolved by Blue Native PAGE
Authors: Kjell Jonas , Rasmusson Allan G. , Larsson Håkan , Widell Susanne
Published: August 2004

With the characterization of the total genomes of Arabidopsis thaliana and Oryza sativa, several putative plasma membrane components have been identified. However, a lack of knowledge at the protein level, especially for hydrophobic proteins, have hampered analyses of physiological changes. To address whether protein complexes may be present in the native membrane, we subjected plasma membranes isolated from Spinacia oleracea leaves to blue‐native polyacrylamide gel electrophoresis (BN‐PAGE). BN‐PAGE is well established in the separation of functional membrane protein complexes from mitochondria and chloroplasts, but a resolved protein complex pattern from PM of eukaryotic cells has previously not been reported. Using this method, protein complexes from Spinacia oleracea PM could be efficiently solubilized and separated, including the highly hydrophobic aquaporin (apparent molecular mass 230 kDa), a putative tetramer of H+‐ATPase, and several less abundant complexes with apparent masses around or above 750 kDa. After denaturation and separation of the complexes into their subunits in a second dimension (SDS‐PAGE), several of the complexes were identified as hydrophobic membrane proteins. Large amounts of protein (up to 1 mg) can be resolved in each lane, which suggests that the method could be used to study also low‐abundance protein complexes, e.g. under different physiological conditions.
Differential Al resistance and citrate secretion in the tap and basal roots of common bean seedlings
Authors: Shen Hong , Yan Xiaolong , Cai Kunzheng , Matsumoto Hideaki
Published: August 2004

The common bean root system is composed of several types of root (e.g. tap, basal, and lateral roots), whose physiological functions may be of great difference. However, we do not know if the root system of common bean differs in organic acid secretion and thus aluminium (Al) resistance. In the present study, the tap and basal roots of three common bean genotypes (i.e. G19842, SQ12 and BAT881) from different origins were compared for their citrate secretion and Al resistance. Grown in a simple solution containing 30 µM Al3+ for 24 h, genotype G19842 maintained 75% relative tap root length [RTRL = (tap root length with Al)/(tap root length without Al)], 48% relative basal root length [RBRL = (basal root length with Al)/ (basal root length without Al)], genotype SQ12 maintained 62% RTRL and 57% RBRL, while BAT881 only maintained 31% RTRL and 19% RBRL, indicating differential sensitivity of bean genotypes and root types to Al stress. The amounts of Al‐induced citrate secretion by the tap/basal roots were 9.8/5.1, 8.2/5.9 and 5.4/4.1 nmol cm−2 FR (fresh root) [12 h]−1 for G19842, SQ12 and BAT881, respectively, indicating that both bean genotypes and root types differ in organic acid secretion. In G19842, the root surface area was 25% higher in tap root apex than that in basal root apex, and the amounts of citrate secretion per unit surface area and per root apex were 29 and 62% higher in tap root apex than those in basal root apex, respectively, suggesting that the higher citrate secretion in the tap root apex could be attributed to the larger surface area and the higher secretion activity. Stronger inhibition of Al‐induced citrate secretion in the basal than tap roots by anthracene‐9‐carboxylic acid, an inhibitor of anion channel and K‐252a, a broad range inhibitor of protein kinase may also imply the differences in the activities of anion channels and K‐252a‐sensitive protein kinases on the plasma membrane between the tap and basal roots, resulting in differential citrate secretion. We propose that the higher Al resistance in the tap root than in basal roots might be attributed to both greater number and higher activity of the anion channels in the former, thus allowing more citrate secretion in this root type.
Further investigation of the roles of auxin and cytokinin in the NH4+‐induced stimulation of nodulation using white clover transformed with the auxin‐sensitive reporter GH3:gusA
Authors: Fei Houman , Vessey J. Kevin
Published: August 2004

Although mineral N (nitrate and ammonium) is believed to have generally negative effects on nodulation in legume–rhizobia symbioses, previous studies have shown that low, static concentrations of ammonium stimulate nodulation in pea, and that this enhancement may be due to an elevation in cytokinin to auxin levels in roots. Here, the effects of ammonium (0.0, 0.1, 0.5 and 2.5 mM) on nodulation and auxin levels were investigated in wild‐type (WT) white clover (Trifolium repens cv. Haifa) and its transformants (lines 38 and 41) which contain the auxin‐sensitive reporter gene (GH3:gusA). The effects of exogenous application (10−10, 10−9 and 10−8 M) of the cytokinin 6‐benzylaminopurine (BAP) were also assessed. Whole‐plant nodulation (nodules plant−1) and dry weight (DW)‐specific nodulation (nodules g−1 root DW) were stimulated (up to 49%) in all white clover lines by 0.1 mM NH4+. This represents the first confirmation of an NH4+‐induced stimulation of DW‐specific nodulation in a species other than pea. At 2.5 mM NH4+, the effect was lost on whole‐plant nodulation and was inhibitory on DW‐specific nodulation. Rhizobial inoculation resulted in a decline in the expression of GH3:gusA in root tips as expected; however, ammonium treatment did not affect GH3 expression in any root zones. Exogenous application of BAP at 10−9 and 10−8 M stimulated whole‐plant and DW‐specific nodulation in wild‐type white clover to a similar degree as treatment with 0.1 mM NH4+. These results support our previous hypothesis that the stimulation of nodulation by low concentrations of ammonium involves the alteration of the ratio of cytokinin to auxin, specifically by increasing cytokinin.
Correlation between chlorophyllide esterification, Shibata shift and regeneration of protochlorophyllide650 in flash‐irradiated etiolated barley leaves
Authors: Rassadina Valentina , Domanskii Vladimir , Averina Natalia G. , Schoch Siegrid , Rüdiger Wolfhart
Published: August 2004

The pigments of etiolated leaves of barley (Hordeum vulgare L.) were analysed during dark periods after flash illumination, and the results were compared with in vivo spectroscopy of the leaves. Pretreatment of the leaves with kinetin slightly stimulated and pretreatment with NaF and anaerobiosis inhibited the esterification of chlorophyllide a (Chlide) at 10–40 min after the flash, whereas the rapid esterification within 30 s after the flash remained unchanged. Irrespective of pretreatment, the amount of esterified pigment was, at any time, identical with the amount of pigment that had shifted its absorption from 684 to 672 nm (Shibata shift). Cycloheximide (CHI) had only a small inhibitory effect on esterification, but drastically inhibited the hydrogenation of geranylgeraniol to phytol, bound to Chlide. The regeneration of long‐wavelength protochlorophyllide a (Pchlide650) was stimulated by kinetin and inhibited by CHI and NaF. During the rapid phase (0–30 s after the flash), the esterification was faster than the regeneration of Pchlide650, and this, in turn, was faster than the formation of photoactive Pchlide. The kinetics changed after pretreatment with 5‐aminolaevulinic acid: regeneration of Pchlide650 was the fastest reaction and the Shibata shift preceded the esterification of Chlide. The results are discussed as pigment exchange reactions at NADPH:protochlorophyllide oxidoreductase (POR; EC 1.6.99.1).
Molecular characterization of a gene encoding a vegetative storage protein (CiVSP) from Cichorium intybus and its expression in the root and shoot in relation to nitrogen status and pathogen resistance
Authors: Richard‐Molard Céline , Brugière Norbert , Moille Murielle , Carrayol Elisa , Limami Anis M.
Published: August 2004

In addition to their putative role in nitrogen storage, some vegetative storage proteins (VSPs) support further roles in biotic and abiotic stress responses. Functions of the 17 kDa VSP from witloof chicory (CiVSP) in N storage and plant resistance to pathogens and its regulation by nitrogen were investigated. The N‐terminal end of this protein was sequenced and the corresponding full‐length cDNA was obtained. The expression of the CiVsp gene was studied in various organs of chicory grown under replete or limited nitrogen supply. A strong expression of CiVsp was observed in both taproot and fine roots of mature plants and seedlings. CiVsp transcripts were also detected in mature leaves, especially in veins. In senescing leaves CiVsp transcripts accumulated concomitantly to a decrease in RbcS transcript abundance and Rubisco small‐subunit degradation. CiVSP protein accumulated significantly only in the subterranean part of the plant during late stages of development. Nitrate limitation caused a reduction in CiVsp mRNA accumulation and a delay in CiVSP storage in the taproot. It is concluded that CiVSP accumulation is regulated at the transcriptional level by N external supply and that the protein is involved in long and short‐term N storage. In silico analysis indicated that CiVSP is highly homologous with several allergens and PR‐10 proteins. Moreover, CiVsp transcript and protein expression were significantly higher in Erwinia carotovora‐resistant chicory inbred lines compared with susceptible lines, suggesting its involvement in chicory resistance to pathogens attack.
Expression of the maize GALACTINOL SYNTHASE gene family: (I) Expression of two different genes during seed development and germination
Authors: Zhao Tian‐Yong , Thacker Richard , Corum J. Willis , Snyder John C. , Meeley Robert B. , Obendorf Ralph L. , Downie Bruce
Published: August 2004

GALACTINOL SYNTHASE (GOLS) acts on UDP‐galactose and myo‐inositol to form galactinol. The production of galactinol is thought to be the committed step in the biosynthesis of the raffinose family oligosaccharides (RFOs), a group of carbohydrates hypothesized to play a number of roles in plant development. Three cDNAs for maize (Zea mays L. inbred line B73) GOLS (ZmGOLS) were identified from the Pioneer Hi‐Bred expressed sequence database. The recombinant proteins from these cDNAs synthesize galactinol. Transcript amounts in developing and germinating seeds were determined using northern blot, ribonuclease protection assays, and multiplex RT‐PCR. ZmGOLS1 mRNA was not detected in any of the tissues examined. ZmGOLS3 transcripts accumulated late during seed development and were present in mature seeds but declined quickly to undetectable amounts following imbibition. Experiments in which germination was interrupted by desiccation, but not by cold, heat, or salt stress, detected ZmGOLS2 transcripts. Soluble sugar amounts and GALACTINOL SYNTHASE activity were determined for developing maize seeds as well as for mature seeds during germination and upon stress during germination. Neither enzyme activity nor galactinol or raffinose quantities could be related to ZmGOLS transcript abundance. Galactinol amounts were most closely related to myo‐inositol quantities during development and early germination. Maturation desiccation appears to be a switch delineating a realm of expression for ZmGOLS3, predominant during seed development, from that of ZmGOLS2, principally associated with post‐imbibition dehydration stress.
Substrate specificity and transport mode of the proton‐dependent amino acid transporter mPAT2
Authors: Foltz Martin , Oechsler Carmen , Boll Michael , Kottra Gabor , Daniel Hannelore
Published: August 2004

The PAT2 transporter has been shown to act as an electrogenic proton/amino acid symporter. The PAT2 cDNA has been cloned from various human, mouse and rat tissues and belongs to a group of four genes (pat1 to pat4) with PAT3 and PAT4 still resembling orphan transporters. The first immunolocalization studies demonstrated that the PAT2 protein is found in the murine central nervous system in neuronal cells with a proposed role in the intra and/or intercellular amino acid transport. Here we provide a detailed analysis of the transport mode and substrate specificity of the murine PAT2 transporter after expression in Xenopus laevis oocytes, by electrophysiological techniques and flux studies. The structural requirements to the PAT2 substrates – when considering both low and high affinity type substrates – are similar to those reported for the PAT1 protein with the essential features of a free carboxy group and a small side chain. For high affinity binding, however, PAT2 requires the amino group to be located in an α‐position, tolerates only one methyl function attached to the amino group and is highly selective for the l‐enantiomers. Electrophysiological analysis revealed pronounced effects of membrane potential on proton binding affinity, but substrate affinities and maximal transport currents only modestly respond to changes in membrane voltage. Whereas substrate affinity is dependent on extracellular pH, proton binding affinity to PAT2 is substrate‐independent, favouring a sequential binding of proton followed by substrate. Maximal transport currents are substrate‐dependent which suggests that the translocation of the loaded carrier to the internal side is the rate‐limiting step.
Neuroserpin Portland (Ser52Arg) is trapped as an inactive intermediate that rapidly forms polymers
Authors: Belorgey Didier , Sharp Lynda K. , Crowther Damian C. , Onda Maki , Johansson Jan , Lomas David A.
Published: August 2004

The dementia familial encephalopathy with neuroserpin inclusion bodies (FENIB) is caused by point mutations in the neuroserpin gene. We have shown a correlation between the predicted effect of the mutation and the number of intracerebral inclusions, and an inverse relationship with the age of onset of disease. Our previous work has shown that the intraneuronal inclusions in FENIB result from the sequential interaction between the reactive centre loop of one neuroserpin molecule with β‐sheet A of the next. We show here that neuroserpin Portland (Ser52Arg), which causes a severe form of FENIB, also forms loop‐sheet polymers but at a faster rate, in keeping with the more severe clinical phenotype. The Portland mutant has a normal unfolding transition in urea and a normal melting temperature but is inactive as a proteinase inhibitor. This results in part from the reactive loop being in a less accessible conformation to bind to the target enzyme, tissue plasminogen activator. These results, with those of the CD analysis, are in keeping with the reactive centre loop of neuroserpin Portland being partially inserted into β‐sheet A to adopt a conformation similar to an intermediate on the polymerization pathway. Our data provide an explanation for the number of inclusions and the severity of dementia in FENIB associated with neuroserpin Portland. Moreover the inactivity of the mutant may result in uncontrolled activity of tissue plasminogen activator, and so explain the epileptic seizures seen in individuals with more severe forms of the disease.
NMR and molecular dynamics studies of an autoimmune myelin basic protein peptide and its antagonist
Authors: Tzakos Andreas G. , Fuchs Patrick , van Nuland Nico A. J. , Troganis Anastasios , Tselios Theodore , Deraos Spyros , Matsoukas John , Gerothanassis Ioannis P. , Bonvin Alexandre M. J. J.
Published: August 2004

Experimental autoimmune encephalomyelitis can be induced in susceptible animals by immunodominant determinants of myelin basic protein (MBP). To characterize the molecular features of antigenic sites important for designing experimental autoimmune encephalomyelitis suppressing molecules, we report structural studies, based on NMR experimental data in conjunction with molecular dynamic simulations, of the potent linear dodecapeptide epitope of guinea pig MBP, Gln74‐Lys75‐Ser76‐Gln77‐Arg78‐Ser79‐Gln80‐Asp81‐Glu82‐Asn83‐Pro84‐Val85 [MBP(74–85)], and its antagonist analogue Ala81MBP(74–85). The two peptides were studied in both water and Me2SO in order to mimic solvent‐dependent structural changes in MBP. The agonist MBP(74–85) adopts a compact conformation because of electrostatic interactions of Arg78 with the side chains of Asp81 and Glu82. Arg78 is ‘locked’ in a well‐defined conformation, perpendicular to the peptide backbone which is practically solvent independent. These electrostatic interactions are, however, absent from the antagonist Ala81MBP(74–85), resulting in great flexibility of the side chain of Arg78. Sequence alignment of the two analogues with several species of MBP suggests a critical role for the positively charged residue Arg78, firstly, in the stabilization of the local microdomains (epitopes) of the integral protein, and secondly, in a number of post‐translational modifications relevant to multiple sclerosis, such as the conversion of charged arginine residues to uncharged citrullines. Flexible docking calculations on the binding of the MBP(74–85) antigen to the MHC class II receptor site I‐Au using haddock indicate that Gln74, Ser76 and Ser79 are MHC II anchor residues. Lys75, Arg78 and Asp81 are prominent, solvent‐exposed residues and, thus, may be of importance in the formation of the trimolecular T‐cell receptor–MBP(74–85)–MHC II complex.
SKELETAL MUSCLE FUNCTION: ROLE OF IONIC CHANGES IN FATIGUE, DAMAGE AND DISEASE
Authors: Allen DG
Published: August 2004

1. Repeated activity of skeletal muscle causes a variety of changes in its properties: muscles become weaker with intense use (fatigue), may feel sore and weak after repeated contractions involving stretch and can degenerate in some disease conditions. The present review considers the role of early ionic changes in the development of each of these conditions.

2. Single fibre preparations of mouse muscle were used to measure ionic changes following activity induced changes in function. Single fibres were dissected with intact tendons and stimulated to produce force. Fluorescent indicators were microinjected into the fibres to allow simultaneous ionic measurements with determination of mechanical performance.

3. One theory to explain muscle fatigue is that fatigue is caused by the accumulation of lactic acid, producing an intracellular acidosis that inhibits the myofibrillar proteins. In contrast, we found that during repeated tetani there was little or no pH change, but that failure of calcium release was a major contributor to fatigue. Currently, it is proposed that precipitation of calcium and phosphate in the sarcoplasmic reticulum contributes to the failure of calcium release.

4. Muscles can be used to shorten and produce force or they can be used to de‐accelerate loads (stretched or eccentric contractions). One day after intense exercise involving stretched contractions, muscles are weak, sore and tender, and this damage can take a week to recover. In this condition, sarcomeres are disorganized and there are increases in resting intracellular Ca2+ and Na+. Recently, we demonstrated that the elevation of Na+ occurs through a stretch‐activated channel that can be blocked by either gadolinium or streptomycin. Preventing the increase in [Na+]i with gadolinium also prevented part of the muscle weakness after stretched contractions.

5. Duchenne muscular dystrophy is a lethal degenerative disease of muscles in which the protein dystrophin is absent. Dystrophic muscles are more susceptible to stretch‐induced muscle damage and the stretch‐activated channel seems to be one pathway for the increases in intracellular Ca2+ and Na+ that are a feature of this disease. We have shown recently that blockers of the stretch‐activated channel can minimize some of the short‐term damage in muscles from the mdx mouse, which also lacks dystrophin. Currently, we are testing whether blockers of the stretch‐activated channels given systemically to mdx mice can protect against some features of the disease.
EFFECT OF ENDOTHELIN‐1 ON REGIONAL KIDNEY BLOOD FLOW AND RENAL ARTERIOLE CALIBRE IN RABBITS
Authors: Denton Kate M , Shweta Amany , Finkelstein Leny , Flower Rebecca L , Evans Roger G
Published: August 2004

1. Medullary blood flow (MBF) is important in the long‐term control of arterial pressure. However, it is unclear which vascular elements regulate MBF.

2. Exogenous endothelin (ET)‐1 decreases cortical more than medullary blood flow. We hypothesized that ET‐1 would therefore constrict afferent (AA) and efferent arterioles (EA) of juxtamedullary glomeruli less than those of cortical glomeruli.

3. Mean arterial pressure, renal blood flow and cortical (CBF) and medullary (MBF) blood flow, via laser‐Doppler flowmetry, were measured before and after intrarenal ET‐1 (2 ng/kg per min; n = 6) or vehicle (n = 6) in anaesthetized rabbits. Kidneys were perfusion fixed, vascular casts formed, lumen diameters measured via scanning electron microscopy and relative resistance calculated.

4. Mean arterial pressure was not significantly affected by ET‐1 infusion. Cortical glomerular arteriole lumen diameters were significantly reduced in the ET‐1‐infused group (AA approximately 30%, EA approximately 18%; PA < 0.01), compatible with the decrease in CBF (42 ± 3%; PGT < 0.01). Juxtamedullary arteriole lumen diameters were also significantly reduced in the ET‐1‐infused group (AA approximately 34%, EA approximately 21%; PA < 0.01); however, MBF did not decrease.

5. In conclusion, our data suggest that juxtamedullary arterioles are not of primary importance in the regulation of MBF because, despite reductions in juxtamedullary arteriole diameters in response to ET‐1, MBF was not decreased.
ARTERIAL STRUCTURAL CHANGES IN HYPERTENSIVE RATS INDUCED BY CAPSAICIN AND SALT LOADING
Authors: Zeng Zhao‐hua , Luo Bi‐hui , Gao Yu‐Jing , Su Cheng‐jian , He Chao‐chu , Li Nina , Lee Robert MKW
Published: August 2004

1. The objective of the present study was to investigate the arterial structural changes in a salt‐sensitive hypertensive rat model induced by treatment with capsaicin.

2. Newborn male Wistar rats were treated with 50 mg/kg capsaicin subcutaneously for 2 days. Control rats were treated with vehicle solution (5% ethanol and 5% Tween 80 in saline). After weaning at 3 weeks, rats were divided into four groups: (i) control with a normal salt diet (0.5% NaCl; CON + NS); (ii) control with a high‐salt diet (4% NaCl; CON + HS); (iii) capsaicin plus normal salt diet (CAP + NS); and (iv) capsaicin plus a high‐salt diet (CAP + HS). Treatment with different salt diets was initiated at 3 weeks of age and lasted for 18 weeks. Tail‐cuff systolic blood pressure (BP) and bodyweight were examined. At the end of the treatment period, blood vessels were prepared by perfusion fixation. Heart weight and vascular dimensions were measured in the thoracic (artery) aorta, renal artery and mesenteric artery.

3. Two weeks after the initiation of the salt diet treatment, BP became significantly higher in the CAP + HS group than in any of the other groups and this difference was maintained until the end of the treatment period.

4. Beginning at 8 weeks after the initiation of the salt diet treatment (11 weeks of age), BP became higher in the CON + HS group than in the CON + NS and CAP + NS groups. Blood pressure was not significantly different between the CON + NS and CAP + NS groups.

5. Media thickness, media thickness to lumen ratio and cross‐sectional area of the aorta, renal artery and mesenteric artery were significantly increased in the CAP + HS group compared with the other groups. Heart weight was also increased in the CAP + HS and CON + HS groups compared with the other groups.

6. Similar structural changes in the blood vessels and heart were also found in the CON + HS group compared with the CON + NS group. Lumen diameter was not altered by the treatments in any of the arteries studied.

7. We conclude that treatment with capsaicin increased the sensitivity of the BP of these rats to salt and this increase in BP is correlated with hypertrophy of the arteries (vascular remodelling) with no change in lumen size. A long‐term high‐sodium load induced hypertension in normal Wistar rats, which was accompanied by cardiovascular hypertrophy.
Efficacy of mizoribine in the treatment of systemic lupus erythematosus in children
Authors: Yoshidome Kouichi , Takei Syuji , Imanaka Hiroyuki , Maeno Nobuaki , Ohkawa Toshiya , Kawano Yoshifumi
Published: August 2004

Abstract
Background
: Mizoribine (MZR) is a novel immunosuppressant developed in Japan. As MZR is reported to be less toxic than other cytotoxic drugs, it is frequently used in Japan in the treatment of adult patients with rheumatoid arthritis or lupus nephritis. The objective of this study was to evaluate the efficacy of MZR in children with SLE. Nine female children with lupus nephritis who had undergone renal biopsy before starting MZR, were involved in this study. Their mean disease duration was 4.8 years at the time MZR treatment was initiated. Patients who had received intensive medications, such as methyl‐prednisolone pulse therapy, intravenous cyclophosphamide pulse therapy, and/or other immunosuppressants, within the 4 months prior to the start of the study, were excluded.

Methods
: Patients treated with 3 mg/kg per day of MZR were monitored every month for up to 1 year. The efficacy of MZR was evaluated by the changes from baseline values of serum C3, serum C4, anti‐dsDNA antibody titer, erythrocyte sedimentation rate (ESR), urinary protein, dosage of prednisolone (PSL), and the sum of the scores defined by these parameters.

Results
: Favorable changes were observed in C3 and ESR after 2 months and 3 months of MZR therapy, respectively. At 3 months of MZR therapy, the sum of scores defined by the parameters for disease activity indicated that MZR was more effective in non‐class IV nephritis patients (n = 5) than in class IV nephritis patients (n = 4) (P = 0.0197). All nine children involved in the study tolerated the MZR therapy well during the study.

Conclusion
: MZR was safe in lupus children, but its efficacy was limited in patients with non‐class IV nephritis. Further study is necessary, in which higher dosages and/or earlier use of MZR is provided to a larger number of children.
Influence of perinatal risk factors on CD3+/TCR αβ and CD3+/TCR γδ lymphocytes in cord blood of preterm neonates
Authors: Królak‐Olejnik Barbara , Mazur Bogdan
Published: August 2004

Abstract
Background
: The aim of the study was the evaluation of CD3+/TCR αβ and CD3+/TCR γδ lymphocytes in the cord blood of the preterm neonates.

Methods
: The study included 26 term neonates as a control group delivered both by spontaneous labor and by cesarean section and 41 preterm neonates: (i) by cesarean section due to abruptio placentae, (ii) by cesarean section due to the danger of intrauterine asphyxia, (iii) by cesarean section due to preterm rupture of membrane (PROM), and (iv) by spontaneous labor. Immunological analysis was performed in the flow cytometer FACScan, using anti‐CD3, anti‐TCR αβ and anti‐TCR γδ monoclonal antibodies from Becton Dickinson (San Jose, California USA).

Results
: It was shown that the way of delivery does not influence the value of CD3+/TCR αβ lymphocytes. A decrease of the percentage and number of CD3+/TCR γδ lymphocytes was found in neonates delivered by elective cesarean section. However, the danger of intrauterine fetal asphyxia, as a reason for preterm delivery, influenced a considerable increase of the number of CD3+/TCR αβ and CD3+/TCR γδ lymphocytes. Perinatal risk factors (abruptio placentae, PROM) were related to the lowest number of CD3+/TCR γδ lymphocytes in the blood of the preterm neonates.

Conclusion
: The obtained results suggest that in spite of considerable immaturity, both a term and preterm neonate is prepared for the immune response and is able to activate cell mechanisms. The precise mechanism that links CD3+/TCR αβ and CD3+/TCR γδ lymphocytes and pathological condition of preterm birth remains unclear.
Antimicrobial susceptibility of respiratory Haemophilus influenzae strains isolated from pediatric respiratory tract infections
Authors: Dong Cao Luong , Naruhiko Ishiwada , Nobue Takeda , Yoichi Kohno
Published: August 2004

Abstract
Background
: Haemophilus influenzae (H. influenzae) is the most frequent bacterial pathogen of respiratory tract infections in children. Detection of antimicrobial susceptibility of H. influenzae is necessary for institution of appropriate antibiotic treatments.

Methods
: A total of 281 strains of H. influenzae isolated from sputum samples of 281 pediatric patients with respiratory tract infections were recruited for study. Antibiotic susceptibility was determined by assessing minimum inhibitory concentrations (MIC) of antimicrobial agents. MIC were measured by utility of Agar dilution susceptibility test.

Results
: Of the total, 38 (13.5%) strains produced β‐lactamase (BLP), 56 (19.9%) strains were β‐lactamase non‐producing, ampicillin resistant (BLNAR). The overall resistant proportion to ampicillin was 33.4%. The data indicated that sulbactam/ampicillin, cefotaxime, ceftriaxone and cefditoren are effective against BLP strains. In addition, a high prevalence of BLNAR H. influenzae strains was identified, with an overall isolation rate of 19.9%. Those strains mainly demonstrated intermediate level to ampicillin (ampicillin‐MIC ≤ 3.13 µg/mL for most of BLNAR strains). However, antimicrobial activities of cefotaxime, ceftriaxone and cefditoren against those strains were slightly diminished. Recovery rate of BLP strains was varied by years, peaking at 19.5%. In particular, a trend of yearly increase in frequency of BLNAR strains was observed.

Conclusion
: Frequency of BLNAR strains causing respiratory infections has been dramatically increasing, which can diminish antibiotic activities of not only ampicillin but also of some cephems.
Value of limited hip abduction in developmental dysplasia of the hip
Authors: Şenaran Hakan , Özdemir H Mustafa , Ögün Tunç Cevat , Kapicioglu M I Safa
Published: August 2004

Abstract
Background
: Developmental dysplasia of the hip (DDH) continues to be missed by routine physical examination in up to 50% of cases. Ultrasound (US) supplementation is the best method of screening for DDH, but the resources required should not be underestimated. Limited abduction of the hip (LHA) in an infant triggers suspicion, and often an urge to treat, in most orthopaedic surgeons and pediatricians alike. This study aimed to document the value of unilateral LHA in the diagnosis and decision making of DDH, and the correlation between LHA and US.

Methods
: In total, 464 infants referred from the pediatrics clinic with LHA, aged between 30 and 120 days, were included in the study.

Results
: Physical examination revealed LHA in 186 (41%) infants, 26 of which were unilateral and 160 were bilateral. US examination showed that 13 (8.1%) patients in the bilateral LHA group and 18 (69.2) patients in the unilateral LHA group, had DDH (total number 31, 7%).

Conclusion
: Unilateral limitation of hip abduction was found to be a sensitive sign for developmental hip dysplasia, but US could be defined once again as the best golden standard before initiating treatment.
Patterns of distribution and abundance of chitons of the genus Ischnochiton in intertidal boulder fields
Authors: GRAYSON J. E. , CHAPMAN M. G.
Published: August 2004

Abstract  Chitons of the genus Ischnochiton are found predominantly on the undersurfaces of boulders, compared with other intertidal or subtidal habitats. They therefore appear to be habitat‐specialists at this scale. This, combined with the fact that boulder fields are relatively sparse compared with other intertidal habitats, makes these animals vulnerable to natural and anthropogenic disturbances. In addition, many species of Ischnochiton are relatively rare and appear to have very patchy abundances, making them likely to have very specific requirements for habitat. We need to understand the habitat requirements in order to manage, conserve and restore disturbed habitats. The present study was carried out at three intertidal boulder fields separated along approximately 200 km of the coast of New South Wales, Australia, centred around Sydney. The boulder fields were representative of those found in this region. The boulders were made of different materials: shale in the north, sandstone in Sydney and quartzitic sandstone in the south. Some boulders in each boulder field were covered by up to 0.4 m of water during low tide. The study showed that the seven species examined were overdispersed among boulders in each of these three intertidal boulder fields. Most boulders did not have associated chitons, but there were very large abundances on a very small number of boulders. Chitons were also overdispersed among boulders that they occupied. These patterns were consistent among shores and among species, even though patterns of abundance were extremely different among different species. These species appear therefore to show specific requirements for habitat at a small spatial scale, using only a small proportion of potential patches of habitat (i.e. boulders) in any place. Extremely patchy patterns of dispersion can be caused by variation in patterns of recruitment, mortality or behavioural responses to habitat or other species. Before performing experiments to investigate such processes, it is useful to test hypotheses of association with habitat using mensurative experiments to identify environmental correlates that might explain the observed patterns. In the present study, sizes of boulders and the associated sessile and mobile assemblages were proposed as mechanisms that could affect dispersion of chitons among boulders. None of these factors, however, showed strong associations with abundances of chitons. The lack of support of these models rules out some features of habitat to which species of Ischnochiton might respond, thereby precluding manipulative experiments involving these features, which are unlikely to be involved in the very patchy patterns of dispersion of species of Ischnochiton.
Sugar and amino acid composition of ant‐attended nectar and honeydew sources from an Australian rainforest
Authors: BLÜTHGEN NICO , GOTTSBERGER GERHARD , FIEDLER KONRAD
Published: August 2004

Abstract  Ants (Hymenoptera: Formicidae) consume a broad spectrum of liquid food sources including nectar and honeydew, which play a key role in their diet especially in tropical forests. This study compares carbohydrates and amino acids from a representative spectrum of liquid sources used by ants in the canopy and understorey of a tropical rainforest in northern Queensland, Australia. Eighteen floral nectars, 16 extrafloral nectars, two wound sap and four homopteran honeydew sources were analysed using high performance liquid chromatography. Wounds comprised flower abscission scars on Normanbya normanbyi L. H. Bailey and bitemarks on Cardwellia sublimis F. Muell. where ants were actively involved in wounding. Discriminant analysis was performed to model differences between food sources in sugar and amino acid concentration and composition. All characteristics varied significantly among plant species. Honeydew contained a broader spectrum of sugars (including melezitose, raffinose, melibiose, lactose and maltose) than nectar (sucrose, glucose, fructose), but certain extrafloral nectars had similar amino acid profiles and, like honeydew sources, were often monopolized by ants. Most common amino acids across the sources were proline, alanine and threonine among 17 α‐amino acids identified. Interspecific variability concealed characteristic differences in sugar and amino acid parameters between nectar, honeydew and wound sap across all plants, but these types differed significantly when found on the same plant. Among all sources studied, only a few flower nectars were naturally not consumed by ants and they were significantly less attended than sugar controls in feeding trials. These nectars did not differ in sugars and amino acids from ant‐attended flower nectars, suggesting the activity of repellents. Apart from these exceptions, variability in amino acids and carbohydrates is proposed to play a key role in ant preferences and nutrition.
Frugivore gape size and the evolution of fruit size and shape in southern hemisphere floras
Authors: LORD JANICE M.
Published: August 2004

Abstract  The present study uses differences among frugivore faunas of the southern hemisphere landmasses to test whether frugivore characteristics have influenced the evolution of fruit traits. Strong floristic similarities exist among southern landmasses; for example, 75% of New Zealand vascular plant genera also have species in Australia. However, plants in Australia and South America have evolved in the presence of a range of mammalian frugivores, whereas those in New Zealand, New Caledonia and the Pacific Islands have not. In addition, the avian frugivores in New Zealand and New Caledonia are generally smaller than those of Australia. If frugivore characteristics have influenced the evolution of fruit traits, predictable differences should exist between southern hemisphere fruits, particularly fruit size and shape. Fruit dimensions were measured for 77 New Zealand species and 31 Australian species in trans‐Tasman genera. New Zealand fruits became significantly more ellipsoid in shape with increasing size. This is consistent with frugivore gape size imposing a selective pressure on fruit ingestability. This result is not a product of phylogenetic correlates, as fruit length and width scaled isometrically for Australian species in genera shared with New Zealand. Within‐genus contrasts between New Zealand and Australian species in 20 trans‐Tasman genera showed that New Zealand species have significantly smaller fruits than their Australian counterparts. Within‐genus contrasts between New Zealand and South American species in nine genera gave the same result; New Zealand species had significantly smaller fruits than their South American counterparts. No difference was found in fruit size or shape between New Zealand and New Caledonia congeneric species from 12 genera. These results are consistent with the broad characteristics of the frugivore assemblage influencing the evolution of fruit size and shape in related species. The smaller‐sized New Zealand frugivore assemblage has apparently influenced the evolution of fruit size of colonizing taxa sometimes within a relatively short evolutionary timeframe.
Role of underground parts of barnyardgrass (Echinochloa crus‐galli (L.) Beauv. var. formosensis Ohwi) in its sensitivity to thenylchlor in soil
Authors: KOBAYASHI KATSUICHIRO , OONAKA NANAKO , SHIM IE‐SUNG
Published: August 2004

In the present study, the phytotoxic activity of top‐soil applied with thenylchlor [2‐chloro‐N‐(3‐methoxy‐2‐thieny)‐2′,6′‐dimethylacetanilide] on the growth of rice (Oryza sativa L.) was dependent on the emergence depth in soil but its activity on barnyardgrass (Echinochloa crus‐galli (L.) Beauv. var. formosensis Ohwi) was only slightly affected by the emergence depth. However, the phytotoxic activity on barnyardgrass and rice was similar irrespective of the different emergence depths in its treatment to all soil layers. Thenylchlor treatment to the mesocotyl of barnyardgrass induced significant inhibition of shoot elongation, whereas the treatment to the coronal root only inhibited the coronal elongation without inhibiting shoot elongation. Absorption and translocation of 14C‐thenylchlor in barnyardgrass were determined in water culture. The different amounts of radioactivity per plant among the treatments to the underground parts were due to the plant part that came in contact with 14C‐thenylchlor. The radioactivity per dry weight was found to be higher in the basal part of the shoot than in its upper part in all treatments to the underground parts. It was suggested that the phytotoxic activity of thenylchlor on the growth of barnyardgrass in soil is induced by its accumulation in the basal part of the shoot through translocation. This primarily occurs after the absorption substantially by the mesocotyl from the herbicide‐treated layer and additionally by other underground parts.
Effect of oocyte transportation time on the clinical results of transport in vitro fertilization/intracytoplasmic sperm injection‐embryo transfer
Authors: TAKANASHI YASUHIRO , ABE YUJI , SHIBUI YUKIHIRO , HANAOKA KANAKO , TAKESHITA NAOKI , MASAKI KAZUO , KUBO HARUMI
Published: August 2004

Background and Aims:  In transport assisted reproductive technology (ART), the time taken to transport oocytes to the main center differs greatly among the satellite facilities and may influence the clinical results.

Methods:  For the conventional in vitro fertilization (IVF) groups in which oocytes were collected at the satellite facilities and transported to the main ART center for insemination and embryo transfer, there were 29 cycles in 27 patients with a transportation time within 60 min (short time transport IVF (ST‐IVF)), 78 cycles in 62 patients with a time between 60 and 120 min (long time transport IVF (LT‐IVF)), and there were 141 cycles in 110 patients at the main ART center (center IVF (C‐IVF)). For the intracytoplasmic sperm injection (ICSI) group, there were 65 ST‐ICSI cycles in 42 patients, 146 LT‐ICSI cycles in 97 patients, and 326 cycles in 238 patients at the main ART center (C‐ICSI).

Results:  The morphologically favorable embryo rate was lower in the ST‐ICSI group (33.8%, P < 0.05) than in the C‐ICSI group (38.1%), and the morphologically poor embryo rate in the LT‐IVF group (38.6%, P < 0.0001) was higher than in the C‐IVF group (26.7%). The rate of embryo transfers resulting in pregnancies was 16.7% in the ST‐ICSI group (P < 0.01) and 17.3% in the LT‐ICSI group (P < 0.001), both less than that of 35.2% for the C‐ICSI group.

Conclusions:  To improve both the morphologically favorable embryo rate and the pregnancy rate in transport ART, it is essential to improve the total quality control at the satellite facilities. (Reprod Med Biol 2004; 3: 123–131)
Current problems of food intake in young women in Japan: Their influence on female reproductive function
Authors: FUJIWARA TOMOKO , NAKATA RIEKO
Published: August 2004

Accumulating evidence suggests that food customs are associated with quality of life in women of the reproductive age. In Japan, dietary limitation for cosmetic purposes, skipping food intake, intake of processed foods and the shift from Japanese to Westernized style food have increased among young women. These changes in food habits can cause inadequate intake of calories, micronutrients, unsaturated fat, phytestrogens and fiber as well as increasing environmental toxins. Furthermore, these food habits increase risk as a result of intake of food additives, anti‐oxidants, processing agents and sweeteners, which have been demonstrated to be harmful to human health. These factors are speculated to not only influence the present lifestyle, but also to induce gynecologic disorders such as dysmenorrhea and irregular menstruation. The adverse effects of these dietary habits on pregnancy outcome and carcinogenesis of breast and ovarian cancers have also been demonstrated. In addition, latent development of organic diseases such as endometriosis, which are accompanied by dysmenorrhea, is a concern under the current nutritional environment in young women. Thus, it is an urgent issue to evaluate the present situation of eating habits in young Japanese women and estimate the influence of these habits on the quality of life including reproductive functions. (Reprod Med Biol 2004; 3: 107–114)
Lipids and lipoprotein(a) concentrations in Pakistani patients with type 2 diabetes mellitus
Authors: Habib S. S. , Aslam M.
Published: August 2004

Aim:  The aim of the present study was to analyze serum lipoprotein(a) [Lp(a)] levels in Pakistani patients with type 2 diabetes mellitus (DM) and to find correlations between clinical characteristics and dyslipidaemias in these patients.

Methods:  Fasting blood samples were analyzed for Lp(a), total cholesterol, triglycerides, low‐density lipoprotein cholesterol (LDL‐c), high‐density lipoprotein cholesterol (HDL‐c), glucose and glycosylated haemoglobin (HbA1c) in 68 Pakistani patients with type 2 DM and 40 non‐diabetic healthy control subjects.

Results:  Lp(a) levels were significantly raised in diabetics as compared to the control group. No correlation of Lp(a) was seen with age, body mass index (BMI), systolic blood pressure (SBP), diastolic blood pressure (DBP) and fasting glucose. There was a positive correlation of BMI to SBP and DBP. There was a significant positive correlation between Lp(a) and total cholesterol and LDL‐c. No correlation of Lp(a) was observed with HDL‐c, triglycerides and glycosylated haemoglobin (HbA1c).

Conclusion:  The present study led us to conclude that serum Lp(a) levels are significantly raised in type 2 DM and have a positive correlation with serum total and LDL‐c levels.
Fluorescence in situ hybridization analysis of matched primary tumour and lymph‐node metastasis of D1 (pT2–3pN1M0) prostate cancer
Authors: Alcaraz Antonio , Corral Juan M. , Ribal María J. , Mallofré Carme , Mengual Lourdes , Carrió Ana , Gil‐Vernet sedó José maría , Villavicencio Humberto
Published: August 2004

OBJECTIVE

To describe the chromosomal numerical changes present in primary prostate tumours and their matched lymph‐node metastases, to identify a clonal cell migration process which could account for the metastatic behaviour.

MATERIALS AND METHODS

Twenty‐eight cases of unsuspected stage D1 (pT2–3pN1M0) prostate cancer were detected among patients who had a radical prostatectomy for clinically localized prostate cancer. Fluorescence in situ hybridization (FISH), using centromeric probes to enumerate chromosomes 7, 8, 10 and 12, was used to assess numerical chromosomal changes. FISH analysis was used on isolated nuclei obtained from matched primary tumours and their lymph node metastases.

RESULTS

Of the 28 suitable cases it was possible to complete the study in 18 pairs of matched tissues; the remainder were excluded because of insufficient tissue or poor preservation of at least one of the tissues. There was cytogenetic change (aneuploidy) in 16 of the 18 primary tumours, the most common being monosomy 8, detected in 14, followed by trisomy 7, in 13 aneuploid tumours. All lymph node metastases were aneuploid by FISH. As in the primary tumours, monosomy 8 and trisomy 7 were the most common cytogenetic alterations, in 13 and 15 of the lymph node tissues. FISH analysis showed a high correlation (83%) in the cytogenetic pattern of changes between the primary tumours and their lymph node metastases. Moreover, a similar number of cells had the most common aneusomies when comparing prostate and the lymph node tissues.

CONCLUSIONS

These results show a similar pattern of cytogenetic alteration in the primary tumour and its lymph node metastasis, characterized by the frequent presence of trisomy 7 and monosomy 8, suggesting that clonal cell selection is not involved in the metastatic process.
Mitochondrial metabolism in the rat during bladder regeneration induced by small intestinal submucosa
Authors: Faramarzi‐Roques Rozbeh , Malgat Monique , Desgrandchamps François , Ballanger Phillipe , Mazat Jean‐Pierre
Published: August 2004

OBJECTIVE

To assess mitochondrial metabolism of bladder tissue induced by small‐intestinal submucosa (SIS), by comparing the mitochondrial enzyme metabolism in this tissue with that in normal bladder tissue and thus evaluate intracellular normality.

MATERIAL AND METHODS

In all, 70 rats were grouped into healthy controls (10), surgical controls with a simple bladder incision (15) and rats treated by partial cystectomy with replacement by the SIS graft (45). At 1, 3 and 6 months the rats were killed, the enzymes of mitochondrial respiratory chain complexes assayed, and the respiration of permeabilized bladder fibres assessed using polarographic analysis.

RESULTS

The enzyme activities of control and treated rats at 3 months were identical. The results from the polarographic analysis of respiration were also similar to that in normal tissue apart from a decrease in the number of mitochondria. Histologically, there was complete regeneration at 6 months.

CONCLUSION

After a phase of inflammation the bladder regenerates after a patch is placed. The new tissue has the same enzymatic and histological features as normal bladder tissue.