bluuebunny/crossref_metadata_2025 huggingface dataset:
The aesthetics of all‐ceramic veneered and monolithic CAD/CAM crowns
Authors: HERRGUTH M. , WICHMANN M. , REICH S.
Published: September 2005

summary The aim of this study was to examine whether crowns fabricated from machinable blocks would achieve acceptable aesthetics and whether these could compete with the aesthetics of restorations obtained by individual layering technique. Fourteen patients, who were to receive single anterior crown restorations, participated in this study. For each person two kinds of crowns were provided: one crown was made with the Cergogold system. The second one was produced in a Cerec machine and was additionally stained. Three independent examiners assessed the aesthetic appearance of crowns fabricated to match each subject's anterior shade. A scale of 1–6 was used to assess the aesthetic adaptation of each crown, with 1 representing excellent characteristics and 3·5 marking the threshold for clinical acceptability. The examiners’ scores were averaged, and the mean values were analysed with the Wilcoxon signed rank test (P ≤ 0·05). Regardless of the fabrication method the crowns were aesthetically acceptable in all 14 patients. The mean values for the layering technique and for the machined restorations did not differ significantly. Within the limits of this study it was documented, that machinable blocks could attain aesthetically satisfying results.
Geometric morphometric assessment of treatment effects of maxillary protraction combined with chin cup appliance on the maxillofacial complex
Authors: CHANG H. P. , LIN H. C. , LIU P. H. , CHANG C. H.
Published: September 2005

summaryThe aim of this study was to investigate the treatment effects on the maxillofacial complex by maxillary protraction combined with chin cup therapy among growing children. An effective geometric morphometric assessment of cephalometric radiographs, using Procrustes analysis and thin‐plate spline analysis, was applied to evaluate shape change in the craniofacial and midfacial configurations of a treated sample of 20 children with skeletal Class III malocclusion. This was compared with matched untreated skeletal Class III controls. Marked treatment induced change involved the maxilla and the mandible. Major deformation consisted of forward advancement of the maxillary complex with negligible rotation of the palatal plane and a forward direction of growth of the mandibular condyle associated with a restriction in sagittal advancement of the chin. Considerable dentoalveolar components contributed to the correction of anterior crossbite. Further detailed study of skeletofacial remodelling in response to maxillary protraction in other skeletal components, including the cranial base and the mandibular complex that contribute to Class III skeletal discrepancies, is warranted.
Conservative treatment of temporomandibular joint osteoarthrosis: intra‐articular injection of sodium hyaluronate
Authors: GUARDA‐NARDINI L. , MASIERO S. , MARIONI G.
Published: September 2005

summary Promising short‐term results in the treatment of temporomandibular joint osteoarthrosis with intra‐articular injections of sodium hyaluronate (SH) have been reported in preliminary studies. The present prospective study compared long‐term outcomes of temporomandibular joint SH injections with those of a conventional non‐surgical treatment (bite‐plane). Data from three groups of 20 patients with degenerative temporomandibular joint disease were considered. Group A underwent one cycle of five injections of 1 mL SH. Group B underwent a bite‐plane treatment for at least 6 months. We considered a control group of 20 patients who refused any treatments. The description of the outcomes was based on objective and subjective parameters after a 6‐month follow‐up. Sodium hyaluronate and bite‐plane treatments significantly improved patients conditions in all considered parameters. No significant differences in outcomes were confirmed by the statistical analysis. The tolerability of SH treatment resulted to be significantly higher. The analysis of results of serial controls in the SH treated group disclosed a significant worsening in pain at rest by comparing 1 and 6 months follow‐up. Sodium hyaluronate infiltration resulted a valid non‐surgical treatment for temporomandibular joint degenerative disease. Five well‐tolerated intra‐articular SH injections achieved equivalent results to those of a 6 months bite‐plane treatment. We did not diagnose any complications of SH intra‐articular injections. Longer time follow‐up is necessary to determine the stability of SH properties.
Low gene flow but high genetic diversity in the threatened Mallorcan midwife toad Alytes muletensis
Authors: KRAAIJEVELD‐SMIT F. J. L. , BEEBEE T. J. C. , GRIFFITHS R. A. , MOORE R. D. , SCHLEY L.
Published: September 2005

We investigated fine‐scale genetic structuring in the rare and vulnerable Mallorcan midwife toad Alytes muletensis using eight polymorphic microsatellite markers. The current range of this amphibian is restricted to some 19 sites of which six are derived from reintroductions, all located in the mountain ranges of Mallorca. We sampled tadpoles from 14 pools covering 10 natural sites and two reintroduction sites for microsatellite DNA analyses. Relatively high levels of genetic variation were found in most pools (HE = 0.38–0.71, allelic richness = 2.6–6.2). Only at one pool has the population recently gone through a bottleneck. Dispersal between pools in different torrents does not occur whereas downstream dispersal between pools within the same torrent does happen at low frequencies. This occasional exchange of individuals does not lead to neighbouring pools in the same torrent being panmictic. This can be concluded because all FST values (0.12–0.53) differ significantly from zero and structure analyses identified neighbouring pools as separate populations. Furthermore, assignment and migration tests showed little exchange between neighbouring pools. If upstream locations or complete torrents go extinct, they are unlikely to be recolonized naturally. For conservation purposes, reintroductions of tadpoles to sites where local extinctions have occurred may therefore be advisable.
Asymmetrical introgression between twoMorusspecies (M. alba,M. rubra) that differ in abundance
Authors: BURGESS K. S. , MORGAN M. , DEVERNO L. , HUSBAND B. C.
Published: September 2005

Asymmetrical introgression is an expected genetic consequence of hybridization when parental taxa differ in abundance; however, evidence for such effects in small populations is scarce. To test this prediction, we estimated the magnitude and direction of hybridization between red mulberry (Morus rubraL.), an endangered species in Canada, and the introduced and more abundant white mulberry (Morus albaL.) using nuclear (randomly amplified polymorphic DNA) and cytoplasmic (chloroplast DNA sequence) markers. Parentage of 184 trees (n = 42 using cpDNA) from four sympatric populations was estimated using a hybrid index and related to six morphological characters and population frequencies of the parental classes. Overall, the frequency of nuclear hybrids was 53.7% (n = 99) and ranged from 43% to 67% among populations. The parental and hybrid taxa differed with respect to all of the morphological traits. Sixty‐seven percent of all hybrids contained more nuclear markers fromM. albathanM. rubra(hybrid indexx̄ = 0.46); among populations, the degree ofM.albabias was correlated with the frequency ofM. alba. In addition, the majority of hybrids (68%) contained the chloroplast genome of white mulberry. These results suggest that introgression is bidirectional but asymmetrical and is related, in part, to the relative frequency of parental taxa.
Gene‐flow patterns in Atlantic and Mediterranean populations of the Lusitanian sea star Asterina gibbosa
Authors: BAUS E. , DARROCK D. J. , BRUFORD M. W.
Published: September 2005

In this study, the population structure of the Lusitanian sea star Asterina gibbosa was assessed using amplified fragment length polymorphism (AFLP). One hundred and twenty‐two AFLP loci were analysed in 159 individuals from eight populations from across the species’ range and revealed high levels of genetic diversity, with all individuals but two harbouring a unique banding pattern. As reported for other marine invertebrates, we found high levels of genetic differentiation between the Atlantic and Mediterranean basins, suggesting that the Strait of Gibraltar represents a major barrier to dispersal for this sea star. Our assignment studies suggest that, in the Atlantic, a measurable degree of gene flow occurs between populations, which could result in the isolation‐by‐distance pattern of differentiation observed in this basin. In contrast, no evidence of contemporary gene flow was found in the Mediterranean, suggesting contrasting patterns of dispersal of Asterina gibbosa in the Atlantic and Mediterranean basins.
Effect of external pH on the growth, photosynthesis and photosynthetic electron transport of Chlamydomonas acidophila Negoro, isolated from an extremely acidic lake (pH 2.6)
Authors: GERLOFF‐ELIAS ANTJE , SPIJKERMAN ELLY , PRÖSCHOLD THOMAS
Published: September 2005

In extremely acidic lakes, low primary production rates have been measured. We assumed that proton stress might explain these observations and therefore investigated the photosynthetic behaviour of a Chlamydomonas species, a main primary producer in acidic lakes, over a range of pH values. Identified as C. acidophila using small subunit rDNA analysis, this species is identical to other isolates from acidic environments in Europe and South America, suggesting a worldwide distribution. Laboratory experiments with C. acidophila, revealed a broad pH‐tolerance for growth and photosynthesis, the lower pH limit lying at pH 1.5 and the upper limit at pH 7. Growth rates at optimum pH conditions (pH 3 and 5) were equal to those of the mesophilic Chlamydomonas reinhardtii. In contrast, photosynthetic rates were significantly higher, suggesting that higher photosynthetic rates compensated for higher dark respiration rates, as confirmed experimentally. Electron transport capacities of PSI and PSII, P700+ re‐reduction times and measurements of PSII fluorescence revealed the induction of alternative electron transport mechanisms, such as chlororespiration, state transitions and cyclic electron transport, only at suboptimal pH values (pH 1.5; 4 and 6–7). The results indicate, that C. acidophila is well adapted to low pH and that the relatively low primary production rates are not a result of pH stress.
Physiological changes in white lupin associated with variation in root‐zone CO2 concentration and cluster‐root P mobilization
Authors: CRAMER M. D. , SHANE M. W. , LAMBERS H.
Published: September 2005

White lupin (Lupinus albus L.) mobilizes insoluble soil phosphorus through exudation of organic acids from ‘cluster’ roots. Organic acid synthesis requires anaplerotic carbon derived from dark CO2 fixation involving PEP‐carboxylase. We tested the hypothesis that variation in root‐zone CO2 concentration would influence organic acid synthesis and thus P mobilization. Root‐zone CO2 concentrations and soil FePO4 concentrations supplied to sand‐grown white lupin (cv. Kiev Mutant) were varied. More biomass accumulated in plants supplied with 360 µL L−1 CO2 to the root‐zone, compared with those aerated with either 100 or 6000 µL L−1 CO2. Increased FePO4 in the sand resulted in greater leaf P concentrations, but root‐zone [CO2] did not influence leaf P concentration. Suppression of cluster‐root development in plants supplied with 100 µL L−1 root‐zone CO2 was correlated with increased leaf [P]. However, at both 360 and 6000 µL L−1 CO2, cluster‐root development was suppressed only at the highest leaf P concentration. Phloem sap [P] was significantly increased by greater [FePO4] in the sand, but was reduced with increased root‐zone [CO2], and this may have triggered increased cluster‐root initiation. Succinate was the major organic acid (carboxylate) in the phloem sap (minor components included malate, citrate, fumarate) and was increased at greater [FePO4], suggesting that this shoot‐derived carboxylate might provide an important source of organic acids for root metabolism. Since cluster root development was inhibited by increasing concentrations of FePO4 in the sand, it is possible that succinate was utilized for the functioning of the root‐nodules.
Analysis of gene expression profiles in Arabidopsis salt overly sensitive mutants sos2−1 and sos3 −1
Authors: KAMEI AYAKO , SEKI MOTOAKI , UMEZAWA TAISHI , ISHIDA JUNKO , SATOU MASAKAZU , AKIYAMA KENJI , ZHU JIAN‐KANG , SHINOZAKI KAZUO
Published: September 2005

Under salt‐stress conditions, Arabidopsis salt overly sensitive (sos) mutants show hypersensitive root growth. The sos gene products are involved in ion transport and signalling processes. To analyse salt‐stress‐responsive genes whose transcripts are regulated by the SOS signalling pathway, we used a 22K Agilent oligoarray and quantitative real‐time polymerase chain reaction analyses. The amounts of transcripts of the major salt‐inducible genes (RD29A/COR78, RD17/COR47, COR15A, KIN1, and RD22) in sos mutants were similar to those in the wild‐type plants. These results indicate that the SOS3/SOS2 signalling pathway is independent of the DRE/CRT, ABRE, and MYC/MYB pathways. In sos2 mutants, the expression levels of 27 and 48 genes were up‐regulated at least double under 2‐h and 5‐h salt stress conditions, respectively. Among the genes regulated by SOS2, we detected several encoding myb family transcription factors, AP2 transcription factors, and some ethylene‐related, auxin‐induced, and defence‐related genes. On the other hand, expression levels of very few genes were different between sos3 mutant and wild‐type plants. This observation indicates that the downstream SOS2 affects the regulation of stress‐responsive genes much more strongly than the upstream SOS3. The gene expression profiles of sos2 and sos3 mutants were not identical. This indicates that SOS2 and SOS3 have individual roles in salt stress responses in addition to common roles.
Near‐term impacts of elevated CO2, nitrogen and fungal endophyte‐infection on Lolium perenne L. growth, chemical composition and alkaloid production
Authors: HUNT MATHEW G. , RASMUSSEN SUSANNE , NEWTON PAUL C. D. , PARSONS ANTHONY J. , NEWMAN JONTHAN A.
Published: September 2005

Carbon dioxide has been rapidly accumulating in the atmosphere and is expected to continue to do so. This accumulation is presumed to have important direct effects on plant growth. The interacting affects of a small increase in CO2 concentration (466 p.p.m., approximately 30% increase from current ambient conditions), nitrogen fertilization and fungal endophyte (Neotyphodium lolii) infection on the growth and chemical composition of perennial ryegrass (Lolium perenne) were investigated. It was found that dry mass production was approximately 50% greater under elevated CO2 than under ambient CO2, but only in conditions of high soil N. High molecular weight carbohydrates and total carbohydrates (LMW + HMW CHO) depended on an interaction between CO2 and endophyte infection. Infected plants contained significantly more carbohydrate than endophyte‐free plants, and the difference was greatest in ambient CO2 conditions. Protein concentrations were also influenced by the interaction between CO2 and endophyte‐infection. Endophyte‐free plants had 40% lower concentrations of soluble protein under elevated CO2 than under ambient CO2, but this CO2 effect on soluble protein was largely absent in endophyte‐infected plants. CO2, endophyte‐infection and nitrogen interacted to influence the total chlorophyll concentration of the grass such that chlorophyll concentration was always lower in elevated CO2 but this decline was much greater in endophyte‐free plants, particularly in conditions of high soil N. In the endophyte‐infected plants, the concentrations of the pyrrolopyrazine alkaloid peramine depended on the interaction between CO2 and N fertilization such that peramine concentrations declined with increasing N at ambient CO2 but remained roughly constant across N levels at elevated CO2. A similar pattern was seen for the ergot alkaloid ergovaline. The biochemical responses of perennial ryegrass to elevated CO2 are clearly modified by the presence of endophytic fungi.
Cell‐wall pectin and its degree of methylation in the maize root‐apex: significance for genotypic differences in aluminium resistance
Authors: ETICHA DEJENE , STASS ANGELIKA , HORST WALTER J.
Published: September 2005

Cell‐wall (CW) pectin content and its degree of methylation in root apices of selected maize cultivars were studied in relation to genotypic Al resistance. Maize cultivars differing in Al resistance were grown in nutrient solution treated with or without Al, and pectin content of the root tips was determined. Control plants did not differ in pectin content in the 5 mm root apex. Al treatment increased the pectin content of the root apex in all cultivars but more prominently in the Al‐sensitive cultivars. Pectin and Al contents in 1 mm root sections decreased from the apex to the 3–4 mm zone. Pectin contents of the apical root sections were consistently higher although significantly different only in the 1–2 mm zone in the Al‐sensitive cv Lixis. Al contents in most root sections were significantly higher in cv Lixis than in Al‐resistant cv ATP‐Y. Localization of pectins by immunofluorescence revealed that Al‐sensitive cv. Lixis has a higher proportion of low‐methylated pectin and thus a higher negativity of the cell wall than Al‐resistant cv ATP‐Y. This is in agreement with the higher Al content and Al sensitivity of cv Lixis. It is concluded that differences in CW pectin and its degree of methylation contribute to genotypic differences in Al resistance in maize in addition to the release of organic acid anions previously reported.
The roles of redox processes in pea nodule development and senescence
Authors: GROTEN KARIN , VANACKER HÉLÈNE , DUTILLEUL CHRISTELLE , BASTIAN FABIOLA , BERNARD STÉPHANIE , CARZANIGA RAFFAELLA , FOYER CHRISTINE H.
Published: September 2005

Nodule senescence is triggered by developmental and environmental cues. It is orchestrated through complex but poorly characterized genetic controls that involve changes in the endogenous levels of reactive oxygen species (ROS) and antioxidants. To elucidate the importance of such redox control mechanisms in pea root nodule senescence, redox metabolites were analysed throughout nodule development in a commercial pea variety (Pisum sativum cv. Phoenix) inoculated with a commercial rhizobial strain (Rhizobium leguminosarum bv. viciae). Although a strong positive correlation between nitrogenase activity and nodule ascorbate and glutathione contents was observed, the progressive loss of these metabolites during nodule senescence was not accompanied by an increase in nodule superoxide or hydrogen peroxide. These oxidants were only detected in nodule meristem and cortex tissues, and the abundance of superoxide or hydrogen peroxide strongly declined with age. No evidence could be found of programmed cell death in nodule senescence and the protein carbonyl groups were more or less constant throughout nodule development. Pea nodules appear to have little capacity to synthesize ascorbate de novo. l‐galactono‐1, 4‐lactone dehydrogenase (GalLDH), which catalyses the last step of ascorbate synthesis could not be detected in nodules. Moreover, when infiltrated with the substrates l‐galactono‐1, 4‐lactone or l‐gulonolactone, ascorbate did not accumulate. These data suggest that ROS, ascorbate and glutathione, which fulfil well recognized, signalling functions in plants, decline in a regulated manner during nodule development. This does not necessarily cause oxidative stress but rather indicates a development‐related shift in redox‐linked metabolite cross‐talk that underpins the development and aging processes.
A portable integrated automatic apparatus for the real‐time monitoring of bioluminescence in plants
Authors: OKAMOTO KAZUHISA , ONAI KIYOSHI , FURUSAWA TAKAYOSHI , ISHIURA MASAHIRO
Published: September 2005

Real‐time monitoring of gene expression by a bioluminescence reporter gene is a powerful method for large‐scale, detailed analysis of gene expression in living cells and large‐scale screening of mutants. We have developed a portable, compact, integrated automatic bioluminescence‐monitoring apparatus that can continuously monitor 960 individual plant seedlings or micro‐organism colonies under uniform light conditions at temperatures up to 50 °C. The apparatus gave reproducible and reliable results for both bioluminescence photon counts and period length of bioluminescence rhythms of Arabidopsis reporter strain. Using the apparatus, we measured bioluminescence rhythms in the thermophilic cyanobacterium Thermosynechococcus at temperature up to 43 °C. We also monitored the expression of the flowering regulator gene CONSTANS in Arabidopsis as bioluminescence in high time resolution under different photoperiodic conditions. The high‐throughput bioluminescence‐monitoring apparatus developed here is a powerful tool for real‐time monitoring of gene expression and gene function.
The influence of light environment on photosynthesis and basal methylbutenol emission from Pinus ponderosa
Authors: GRAY DENNIS W. , GOLDSTEIN ALLEN H. , LERDAU MANUEL T.
Published: September 2005

Methylbutenol is a 5‐carbon alcohol that is produced and emitted by several species of pine in western North America, and may have important impacts on the tropospheric chemistry of this region. In the present study the response of methylbutenol basal emission rate (measured at a constant light intensity of 1500 µmol m−2 s−1 and temperature of 30 °C) to the light and temperature conditions of the growth environment was examined, using field‐grown plants shielded with shade cloth of various densities. Methylbutenol basal emission rates increased linearly with the temperature of the growth environment but did not respond to the shading of foliage during growth and development. Both photosynthesis and basal methylbutenol emission rate declined in older needles; however, these declines appear to result from parallel but independent processes and not from basal MBO emission rate directly tracking photosynthetic rates. Older needles did not occupy cooler microenvironments within the canopy; and thus differing thermal microenvironment could not explain the reduced MBO emission in older needles.
Regulation of plant defense responses in Arabidopsis by EDR2, a PH and START domain‐containing protein
Authors: Tang Dingzhong , Ade Jules , Frye Catherine A. , Innes Roger W.
Published: September 2005

We have identified an Arabidopsis mutant that displays enhanced disease resistance (edr2) to the biotrophic powdery mildew pathogen Erysiphe cichoracearum. Inhibition of fungal growth on edr2 mutant leaves occurred at a late stage of the infection process and coincided with formation of necrotic lesions approximately 5 days after inoculation. Double‐mutant analysis revealed that edr2‐mediated resistance is suppressed by mutations that inhibit salicylic acid (SA)‐induced defense signaling, including npr1, pad4 and sid2, demonstrating that edr2‐mediated disease resistance is dependent on SA. However, edr2 showed normal responses to the bacterial pathogen Pseudomonas syringae pv. tomato strain DC3000. EDR2 appears to be constitutively transcribed in all tissues and organs and encodes a novel protein, consisting of a putative pleckstrin homology (PH) domain and a steroidogenic acute regulatory protein‐related lipid‐transfer (START) domain, and contains an N‐terminal mitochondrial targeting sequence. The PH and START domains are implicated in lipid binding, suggesting that EDR2 may provide a link between lipid signaling and activation of programmed cell death mediated by mitochondria.
Sensitive detection and localization of hydroxyl radical production in cucumber roots and Arabidopsis seedlings by spin trapping electron paramagnetic resonance spectroscopy
Authors: Renew Susannah , Heyno Eiri , Schopfer Peter , Liszkay Anja
Published: September 2005

As reactive oxygen species are important for many fundamental biological processes in plants, specific and sensitive techniques for their detection in vivo are essential. In particular, the analysis of hydroxyl radical (OH•) formation in biological reactions has rarely been attempted. Here, it is shown that spin trapping electron paramagnetic resonance (EPR) spectroscopy allows the detection and quantitative estimation of OH• production in vivo in one single cucumber seedling root. It is possible to localize the OH• production site to the growth zone of the root by varying the position of the intact seedling inside the resonator cavity of the EPR spectrometer. Moreover, the demonstration of impaired OH• formation in the root of the Arabidopsis mutant rhd2 impaired in a superoxide‐producing Nicotimamide adenine dinucleotide phosphate (NADPH) oxidase has been accomplished. Spin trapping EPR provides a valuable tool for analyzing the production of OH•in vivo with high resolution in small tissue samples.
ELF4 is a phytochrome‐regulated component of a negative‐feedback loop involving the central oscillator components CCA1 and LHY
Authors: Kikis Elise A. , Khanna Rajnish , Quail Peter H.
Published: September 2005

Evidence has been presented that a negative transcriptional feedback loop formed by the genes CIRCADIAN CLOCK ASSOCIATED (CCA1), LATE ELONGATED HYPOCOTYL (LHY) and TIMING OF CAB (TOC1) constitutes the core of the central oscillator of the circadian clock in Arabidopsis. Here we show that these genes are expressed at constant, basal levels in dark‐grown seedlings. Transfer to constant red light (Rc) rapidly induces a biphasic pattern of CCA1 and LHY expression, and a reciprocal TOC1 expression pattern over the first 24 h, consistent with initial induction of this synchronous oscillation by the light signal. We have used this assay with wild‐type and mutant seedlings to examine the role of these oscillator components, and to determine the function of ELF3 and ELF4 in their light‐regulated expression. The data show that whereas TOC1 is necessary for light‐induced CCA1/LHY expression, the combined absence of CCA1 and LHY has little effect on the pattern of light‐induced TOC1 expression, indicating that the negative regulatory arm of the proposed oscillator is not fully functional during initial seedling de‐etiolation. By contrast, ELF4 is necessary for light‐induced expression of both CCA1 and LHY, and conversely, CCA1 and LHY act negatively on light‐induced ELF4 expression. Together with the observation that the temporal light‐induced expression profile of ELF4 is counter‐phased to that of CCA1 and LHY and parallels that of TOC1, these data are consistent with a previously unrecognized negative‐feedback loop formed by CCA1/LHY and ELF4 in a manner analogous to the proposed CCA1/LHY/TOC1 oscillator. ELF3 is also necessary for light‐induced CCA1/LHY expression, but it is neither light‐induced nor clock‐regulated during de‐etiolation. Taken together, the data suggest (a) that ELF3, ELF4, and TOC1 all function in the primary, phytochrome‐mediated light‐input pathway to the circadian oscillator in Arabidopsis; and (b) that this oscillator consists of two or more interlocking transcriptional feedback loops that may be differentially operative during initial light induction and under steady‐state circadian conditions in entrained green plants.
Predictors of diabetic end‐stage renal disease in Japan
Authors: ISEKI KUNITOSHI
Published: September 2005

SUMMARY:  Diabetes mellitus (DM) has been the leading cause of incident dialysis in Japan since 1998, according to the Japanese Society for Dialysis Therapy (JSDT). In particular, the number of male DM dialysis patients is increasing. DM is becoming a worldwide epidemic in both developed and developing countries. Strategies to detect individuals at high‐risk of developing CKD and end‐stage renal disease (ESRD) are needed that can be implemented on a population‐basis.

Among the commonly measured variables, dipstick urinalysis (proteinuria, haematuria), blood pressure, serum creatinine, body mass index (BMI), and serum uric acid are significant predictors of ESRD. Recently, we evaluated the effect of DM as a risk factor of developing ESRD. DM was diagnosed when the fasting plasma glucose (FPG) was 126 mg/dL or more in participants (n = 78529) of the 1993 screening program in Okinawa. The prevalence of DM was 5.2%. The odds ratio (95% CI) of DM for developing ESRD was 3.098 (1.738–5.525, P = 0.0001) after adjusting for possible confounding variables. Early detection and treatment of DM might prevent DM‐related ESRD. We examined 7125 non‐DM screenees who underwent a 1‐day health check between April 1997 and March 1998. They were followed‐up until March 2000 to determine whether they developed DM. Over the 2 years, the cumulative incidence of DM was 2.3%, 2.9% in men and 1.3% in women. Proteinuria was the most robust predictor of the development of DM; the adjusted relative risk (95% CI) was 1.90 (1.14–3.17). Obesity, per se, is also recognized as a risk factor for developing proteinuria. The higher the BMI, the higher the risk of developing ESRD; the adjusted odds ratio (95% CI) was 1.273 (1.121–1.446, P = 0.0002) for men. Other than being overweight (BMI = 25.0 kg/m2), a smoking habit was a significant predictor of developing proteinuria.

The prevalence of obesity and DM is increasing in Japan. It is possible that the impact of obesity and complications of DM are different among races and ethnicities. Public relations regarding the risk of DM and its complications are especially important in Asian countries. Asians have more fat than non‐Asians, even at the same BMI levels. Knowledge of the predictors of DM‐ESRD is crucial as a first step toward prevention. Consistent with this notion, initiatives on the management of CKD and ESRD were recently organized in Japan and internationally.
Plasminogen activator inhibitor‐1 and diabetic nephropathy
Authors: LEE HI BAHL , HA HUNJOO
Published: September 2005

SUMMARY:  Diabetic nephropathy is characterized by excessive accumulation of extracellular matrix (ECM) in the kidney. Decreased ECM degradation as well as increased ECM synthesis plays an important role in ECM remodeling that favours tissue fibrosis. Plasminogen activator (PA)/plasmin/PA inhibitor (PAI) system is involved in ECM degradation and PAI‐1 plays a critical role in ECM remodeling in the kidney. Normal human kidneys do not express PAI‐1 but PAI‐1 is overexpressed in pathologic conditions associated with renal fibrosis including diabetic nephropathy. Reactive oxygen species mediate PAI‐1 up‐regulation in renal cells cultured under high glucose, hypoxia, and TGF‐β1. Recent studies utilizing PAI‐1 deficient mice suggest that PAI‐1 induce ECM deposition in diabetic kidney through increased ECM synthesis by TGF‐β1 up‐regulation as well as through decreased ECM degradation by suppression of plasmin and MMP‐2 activity.
Podocyte hypertrophy in diabetic nephropathy
Authors: KIM NAM HO
Published: September 2005

SUMMARY:  The development of irreversible renal changes in diabetes mellitus, such as glomerulosclerosis and tubulointerstitial fibrosis, are always preceded by the early hypertrophic processes in the glomerular and tubular compartment. However, the role of hypertrophy of podocyte in the diabetic nephropathy have not been fully elucidated yet. Observation came from a cross sectional study in diabetic Pima Indians suggests that subjects with clinical nephropathy had fewer podocytes per glomerulus than those without nephropathy. Since podocytes are thought to be incapable of replication, this observation suggests that podocyte loss, or perhaps a low podocyte number per glomerulus, contributes to the development and progression of diabetic glomerulosclerosis. Podocyte hypertrophy caused by high glucose concentration leads to podocyte loss and is a new insight of pathogenesis of diabetic nephropathy; and it also provides us with new therapeutic strategies in diabetic nephropathy.
Genetics of diabetic nephropathy in type 2 DM: candidate gene analysis for the pathogenic role of inflammation
Authors: LEE SANG‐HO , LEE TAE WON , IHM CHUN‐GYOO , KIM MYUNG JAE , WOO JEOUNG‐TAEK , CHUNG JOO‐HO
Published: September 2005

SUMMARY:  Hypertension, poor glycemic control and albuminuria are well known risk factors for diabetic nephropathy, but these factors do not explain all of the inter‐individual variabilities in the rate of progression to kidney failure. Recent evidence showed that genetic predisposition affected the hyperglycemia‐induced nephrotoxicity in patients with type 2 diabetes mellitus (DM). We reviewed the present state of knowledge concerning the relationship between genetics and diabetic nephropathy in type 2 DM. However, the results are inconclusive and the genetic determinants of diabetic nephropathy are not fully understood. In addition, genetic background of nephropathy in type 2 DM was thought to be more complex than in type 1 DM. Recent studies suggested that the inflammation would be an essential component of type 2 DM and its complications. We postulated that increased systemic and/or intrarenal inflammation in high glucose milieu is important in the pathogenesis of nephropathy in patients with type 2 DM. To investigate the impact of inflammation on diabetic nephropathy, we studied several polymorphisms in genes encoding inflammatory cytokine and chemokine in patients with type 2 DM. Among them, −511 C/T in interleukin‐1β (IL‐1β), tandem repeat in IL‐1 receptor antagonist (IL‐1Ra), −308 G/A in tumour necrosis factor‐α (TNF‐α) were significantly associated with an increased risk of kidney failure. In addition, some of them were remarkably different from those previously reported in the NCBI or literature based on the western population.

Our results suggest that inflammation could play a pathogenic role in diabetic nephropathy in type 2 DM. A better understanding of genetic factors predisposing to diabetic nephropathy would not only help to identify diabetic patients at risk, but also be helpful to unveil the pathogenesis of DN.
Role of aldosterone in diabetic nephropathy
Authors: CHA DAE RYONG , KANG YOUNG SUN , HAN SANG YOUB , JEE YI HWA , HAN KUM HYUN , KIM HYOUNG KYU , HAN JEE YOUNG , KIM YOUNG SIK
Published: September 2005

SUMMARY:  In the last 10 years, many studies have focused on the non‐classical action of aldosterone. One of the most important new aspects of aldosterone is its pathogenic role as proinflammatory and profibrotic molecules. It has been reported that aldosterone induces myocardial fibrosis and vascular inflammation through up‐regulation of various proinflammatory and profibrotic cytokines. We investigated the effect of aldosterone and spironolactone, which is a non‐selective mineralocorticoid receptor antagonist, on monocyte chemoattractant peptide (MCP‐1) and collagen synthesis in cultured mesangial and tubular epithelial cells. In addition, to evaluate the effect of spironolactone on diabetic nephropathy, we used Otsuka Long‐Evans Tokushima Fatty (OLETF) rats which are known type 2 diabetic animal models. Spironolactone treatment did not induce any significant change in blood glucose levels and blood pressure. However, spironolactone therapy significantly inhibited urinary albumin and MCP‐1 excretion. Spironolactone treatment also suppressed renal mRNA expression for MCP‐1, macrophage migration inhibitory factor (MIF) as well as intrarenal protein synthesis for ED‐1 and MIF. Morphologically, spironolactone treatment significantly prevented glomerulosclerosis, collagen deposition and connective tissue growth factor (CTGF) expression in diabetic rats. In cultured cell experiments, aldosterone directly increased the MCP‐1, collagen secretion and spironolactone treatment abolished aldosterone‐induced MCP‐1 and collagen synthesis. Surprisingly, aldosterone treatment did not induce any significant change in TGFβ1 gene transcription. Finally, we found that NF‐kB activity was increased after stimulation with aldosterone and spironolactone therapy inhibited their activation. In addition, prior treatment with pyrrolidine dithiocarbamate (PDTC), which is a NF‐KB inhibitor, inhibited aldosterone‐induced MCP‐1 protein secretion. These results suggest that aldosterone blockade could play a role in preventing the progression of diabetic nephropathy via anti‐inflammatory and antifibrotic mechanisms.
Clinical characteristics of mutation carriers in a large family with glucokinase diabetes (MODY2)
Authors: Shehadeh N. , Bakri D. , Njølstad P. R. , Gershoni‐Baruch R.
Published: September 2005

Aims  To investigate the prevalence and clinical characteristics of heterozygotes of the glucokinase gene mutations G264S and IVS8+2 in the extended pedigree of two patients with permanent neonatal diabetes as a result of glucokinase deficiency (IVS8+2 homozygosity and IVS8+2/G264S compound heterozygosity).

Methods  Eighty‐eight first, second and third degree family members of the two patients with permanent neonatal diabetes were genotyped. Clinical, laboratory and historical data were collected via chart reviews.

Results  Thirty‐one IVS8+2 and three G264S heterozygotes were identified. Of these, 18/34 (52.9%) had diabetes and 9/34 (26.5%) impaired fasting glucose (IFG), compared with 1/54 (1.9%) with diabetes and 2/54 (3.7%) with IFG in the non‐carrier group. Odds ratio for heterozygotes was 70.4 (95% CI 16.9–293.5 and P < 0.001). Mean body mass index of heterozygotes (> 18 years of age) who had diabetes was 27.1 ± 2.66, compared with 23.18 ± 4.72 for heterozygotes with normal glucose levels (P < 0.05). While none of the non‐carrier women had gestational diabetes, eight of the 10 heterozygotes developed gestational diabetes. Inheritance of a glucokinase mutation by the fetus from a carrier mother resulted in a significant reduction in birthweight (3600 ± 570 vs. 2970 ± 390grams, P < 0.05).

Conclusions  These data support the association between carriage of GCK gene mutations, G264S and IVS8+2, and the development of diabetes, impaired fasting glucose and reduced birthweight. Moreover, in heterozygotes, a clear correlation between body mass index and the development of diabetes was observed. These findings underline the need for surveillance and prevention in individuals at risk.
Increased plasma markers of inflammation and endothelial dysfunction and their association with microvascular complications in Type 1 diabetic patients without clinically manifest macroangiopathy
Authors: Targher G. , Bertolini L. , Zoppini G. , Zenari L. , Falezza G.
Published: September 2005

Aims  To evaluate whether plasma biomarkers of inflammation and endothelial dysfunction differed in Type 1 diabetic patients as compared with those in non‐diabetic subjects, and to examine the association of these biomarkers with early stages of microvascular complications.

Methods  Plasma biomarkers of inflammation [fibrinogen, hs‐C‐reactive protein (hs‐CRP)] and endothelial dysfunction [von Willebrand factor (v‐WF), intercellular adhesion molecule‐1, plasminogen activator inhibitor‐1 (PAI‐1) activity] were measured in 88 non‐smoking young patients with Type 1 diabetes without clinical macrovascular disease and in 40 healthy controls.

Results  Plasma levels of hs‐CRP, fibrinogen, v‐WF, soluble intracellular adhesion molecule‐1 (sICAM‐1) and PAI‐1 activity were markedly higher (P < 0.01 or less) in Type 1 diabetic patients than in healthy controls; these results were essentially unchanged when healthy controls were compared with patients without complications. After stratification by microvascular complication status, plasma biomarkers of inflammation and endothelial dysfunction were significantly increased in those with more advanced disease compared with those with early complications or without complications, respectively. However, while the significant differences in these biomarkers were little affected by adjustment for sex, age, BMI and blood pressure values, they were totally abolished after additional adjustment for diabetes duration and glycaemic control.

Conclusions  These results indicate that in Type 1 diabetes there is a subclinical, chronic inflammation which is, at least partly, independent of clinically manifest macro‐ and microvascular complications, smoking or other traditional cardiovascular risk factors; this subclinical inflammation is closely correlated to the magnitude and duration of hyperglycaemia.
Impairment of cardiac function in hypertensive patients with Type 2 diabetes: a LIFE study
Authors: Hildebrandt P. , Wachtell K. , Dahlöf B. , Papademitriou V. , Gerdts E. , Giles T. , Oikarinen L. , Tuxen C. , Olsen M. H. , Devereux R. B.
Published: September 2005

Aims  Type 2 diabetic patients with hypertension have an increased left ventricular (LV) mass and impaired cardiac function compared to hypertensive patients without diabetes. However, it is unknown if the impaired cardiac function can be explained solely by LV hypertrophy, or is independently related to diabetes.

The aim of the present study was to compare LV function between diabetic and non‐diabetic hypertensive patients with electrocardiographic LV hypertrophy.

Methods  In 937 patients participating in the LIFE echocardiographic substudy, all echocardiograms were centrally evaluated by a core reading centre measuring LV mass, systolic and diastolic LV function. Known diabetes was present in 105 patients.

Results  Left ventricular mass was similar in diabetic and non‐diabetic patients. Endocardial systolic LV function, estimated by LV ejection fraction, was reduced and indices of midwall systolic LV function were impaired in the diabetic patients. Diastolic LV filling pattern was impaired and arterial stiffness, measured by pulse pressure/stroke index, was increased in diabetic patients.

Conclusions  Systolic and diastolic LV function in hypertensive patients with electrocardiographic LV hypertrophy and diabetes are impaired independent of LV mass, most likely reflecting the adverse effects of diabetes per se.
Diagnostic screening of NEUROD1 (MODY6) in subjects with MODY or gestational diabetes mellitus
Authors: Sagen J. V. , Baumann M. E. , Salvesen H. B. , Molven A. , Søvik O. , Njølstad P. R.
Published: September 2005

Aims  Diagnostic screening of NEUROD1 in patients with maturity‐onset diabetes of the young (MODY) without mutations in the known MODY‐genes (MODYX) and in subjects diagnosed with gestational diabetes mellitus.

Methods  Direct sequencing of NEUROD1 was performed in (i) 73 probands with clinical MODY without mutations in hepatocyte nuclear factor (HNF)‐4α (MODY1), glucokinase (MODY2) and hepatocyte nuclear factor (HNF)‐1α (MODY3), and (ii) 51 subjects diagnosed with gestational diabetes. Control material consisted of 105 anonymous blood donors.

Results  Mean age at diagnosis of diabetes was 22 and 30 years in the MODYX patients and gestational diabetes mellitus subjects, respectively. Mean fasting blood glucose (9.6 ± 4.3 vs. 5.7 ± 1.0 mml/l) as well as glycosylated haemoglobin (8.2 ± 2.4 vs. 6.0 ± 0.6%) were higher in the MODYX patients than subjects with gestational diabetes. NEUROD1 mutations were not detected in our two study groups. Three previously reported polymorphisms were found: Ala45Thr, Pro197His and IVS1 −32 nt C>T. The amino acid substitution serine to cysteine in codon 29 (designated Ser29Cys) was detected in one out of 105 control subjects. As the control material consisted of anonymous blood donors, we were prevented from investigation of possible co‐segregation between the sequence variant Ser29Cys and diabetes mellitus.

Conclusions  As we found no NEUROD1 mutations, diagnostic screening for this gene is not warranted in Norwegian MODYX patients. Our study also suggests that NEUROD1 is not a candidate gene in gestational diabetes mellitus (GDM). The sequence variant Ser29Cys was identified in one anonymous DNA sample, but we were prevented from studying possible co‐segregation with diabetes mellitus.
Effects of liraglutide (NN2211), a long‐acting GLP‐1 analogue, on glycaemic control and bodyweight in subjects with Type 2 diabetes
Authors: Feinglos M. N. , Saad M. F. , Pi‐Sunyer F. X. , An B. , Santiago O. ,   on behalf of the Liraglutide Dose‐Response Study Group
Published: September 2005

Aims  Liraglutide (NN2211) is a long‐acting GLP‐1 analogue, with a pharmacokinetic profile suitable for once‐daily administration. This multicentre, double‐blind, parallel‐group, double‐dummy study explored the dose–response relationship of liraglutide effects on bodyweight and glycaemic control in subjects with Type 2 diabetes.

Methods  Subjects (BMI 27–42 kg/m2) with Type 2 diabetes who were previously treated with an OAD (oral anti‐diabetic drug) monotherapy (69% with metformin), and had HbA1c ≤ 10% were enrolled. After a 4‐week metformin run‐in period, 210 subjects (27–73 years, 60% female) were randomised to receive liraglutide (0.045–0.75 mg) once daily or continued on metformin 1000 mg b.d. for 12 weeks.

Results  Mean baseline values for the six treatment groups ranged from 6.8 to 7.5% for HbA1c, and 8.06–9.44 mmol/l (145–170 mg/dl) for fasting plasma glucose. After 12‐week treatment, a weight change of −0.05 to −1.9% was observed for the six treatment groups. Mean HbA1c changes from baseline for 0.045, 0.225, 0.45, 0.6, 0.75 mg liraglutide and metformin were +1.28%, +0.86%, +0.22%, +0.16%, +0.30% and +0.09%, respectively. No significant differences in HbA1c were observed between liraglutide and metformin groups at the three highest liraglutide dose levels (0.45, 0.6 and 0.75 mg). The lowest two liraglutide doses (0.045 mg and 0.225 mg) were not sufficient to maintain the fasting plasma glucose values achieved by metformin. No major hypoglycaemic episodes were reported. Episodes of nausea and/or vomiting were reported by 11 patients (6.3%) receiving liraglutide and three (8.8%) receiving metformin.

Conclusions  Once‐daily liraglutide improved glycaemic control and weight, in a comparable degree to metformin. Liraglutide appeared to be safe and generally well tolerated. Higher doses of liraglutide merit study in future clinical trials.
Sibutramine improves insulin sensitivity without alteration of serum adiponectin in obese subjects with Type 2 diabetes
Authors: Hung Y.‐J. , Chen Y.‐C. , Pei D. , Kuo S.‐W. , Hsieh C.‐H. , Wu L.‐Y. , He C.‐T. , Lee C. H. , Fan S. C. , Sheu W. H.‐H.
Published: September 2005

Aim  To evaluate the effect of sibutramine on weight loss, insulin sensitivity and serum adiponectin levels in obese patients with Type 2 diabetes.

Methods  This study is a randomized, double‐blind, placebo‐controlled parallel comparison study of sibutramine 15 mg/day and placebo. Forty‐eight eligible obese patients with Type 2 diabetes (age between 30 and 75 years with body mass index ≥ 27 kg/m2) were randomly assigned to receive either placebo (n = 24) or sibutramine (15 mg/day) (n = 24) for 6 months. Fifteen subjects in each group underwent meal tests and modified insulin suppression tests before and after 6 months’ treatment.

Results  After 6 months of sibutramine treatment statistically significant changes from baseline were observed for body weight (85.4 ± 2.5 vs. 82.9 ± 2.4 kg, P < 0.005) and body mass index (32.0 ± 0.7 vs. 31.4 ± 0.6 kg/m2, P < 0.05) without a significant alteration of waist–hip ratio (W/H), blood pressure, heart rate, glycaemic parameters or lipid profiles. The steady‐state plasma glucose (SSPG) level during the modified insulin suppression test was significantly reduced in the sibutramine group (17.33 ± 2.92 vs. 14.29 ± 4.19 mmol/l, P < 0.05) despite similar steady‐state plasma insulin (SSPI) concentrations. In addition, serum adiponectin and C‐reactive protein (CRP) levels remained unchanged, although modest weight reduction was achieved after sibutramine treatment. There were also no significant correlations between changes in serum adiponectin and reduction of SSPG or body weight. Daily ambient plasma insulin and glucose concentrations in response to a test meal were not significantly different in subjects receiving sibutramine treatment.

Conclusions  We conclude that treatment with sibutramine 15 mg once daily effectively reduces weight and enhances insulin sensitivity without alteration of serum adiponectin levels in obese patients with Type 2 diabetes.