bluuebunny/crossref_metadata_2025 huggingface dataset:
Effects of smoking on local delivery of controlled‐release doxycycline as compared to scaling and root planing
Authors: Ryder Mark I. , Pons Barbara , Adams Donald , Beiswanger Bradley , Blanco Valerie , Bogle Gary , Donly Kevin , Hallmon William , Hancock E. Brady , Hanes Philip , Hawley Charles , Johnson Lonnie , Wang Hom‐Lay , Wolinsky Lawrence , Yukna Raymond , Polson Alan , Carron Graham , Garrett Steven
Published: October 2003

Abstract. This paper examines the effects of smoking on the treatment outcomes of two nonsurgical therapies: (1) scaling and root planing alone (SRP) or (2) controlled‐release of subgingivally delivered doxycycline hyclate in a polylactic acid based polymer gel. Subjects from 2 9‐month multicenter studies were classified as nonsmokers (never smoked: 100 subjects), former smokers (137 subjects), and current smokers (≥10 cigarettes/day: 121 subjects). Clinical parameters were analyzed for treated sites with baseline probing depths ≥5 mm and for a subset of treated sites with baseline probing depths of ≥7 mm. Clinical parameters (plaque levels, clinical attachment levels, pocket depths, and bleeding on probing) were analyzed at baseline, 4, 6, and 9 months. In the doxycycline treated group in general, there were neither marked significant differences in clinical attachment gain nor differences in probing depth reduction among the 3 smoking groups. On the other hand, in the scaling and root planing treated group in general, there were significant differences in clinical attachment gain and pocket depth reduction, with non‐smokers responding better than former smokers and current smokers at 6 and 9 months. These differences in clinical response between scaling and root planing alone versus controlled‐release of locally‐delivered doxycycline hyclate among these 3 smoking groups are discussed in relation to treatment implications for smokers.
Quantification of root surface plaque using a new 3‐D laser scanning method
Authors: Yeganeh S. , Lynch E. , Jovanovski V. , Zou L.
Published: October 2003

Abstract. There are no published reports in the literature objectively quantifying thickness of plaque on teeth. The aim of this study was to quantify plaque on a tooth surface and assess if this quantification correlates with a clinical index of plaque from each of 51 patients. Patients were instructed not to perform any oral hygiene on the day of the assessment. The Silness and Löe plaque index was scored and replicas were scanned using a co‐ordinate measuring machine (CMM) and laser scanning probe. A replica was obtained from this surface before and after toothbrushing. Plaque adjacent to the gingival margin had a mean thickness of 0.106±0.118 mm (mean±SD) whilst mean plaque thickness 250 μm from the gingival margin was 0.053±0.052 mm (mean±SD). There was a significant correlation between the plaque index and the plaque thickness (p≤0.002). The finding that plaque is present in the greatest amount adjacent to the gingival margin supports a previously reported hypothesis that primary root carious lesions (PRCL's) may initiate adjacent to the gingival margin. This method quantifies plaque thickness on exposed root surfaces which correlates with the plaque index as well as illustrating how the morphological characteristics of teeth, gingivae and plaque can be studied in vivo from replicas recorded.
The relationship between the classification of Scottish ground beetle assemblages (Coleoptera, Carabidae) and the National Vegetation Classification of British plant communities
Authors: Blake Shona , McCracken David I. , Eyre Michael D. , Garside Adam , Foster Garth N.
Published: October 2003

A sufficiently large body of knowledge on British ground beetle (Carabidae) communities now exists to allow investigation of whether habitats may be classified or described on the basis of their ground beetle communities, in the same way that the National Vegetation Classification (NVC) describes British plant communities. A data set of ground beetle abundances from pitfall traps at 481 sites in a range of natural, semi‐natural and agricultural habitats throughout Scotland was available for analysis. Multivariate analysis (detrended correspondence analysis and fuzzy cluster analysis) was carried out on proportional catch data of 156 species of ground beetle from 444 of these sites and the results related to the NVC of the sites.

Initial analysis classified the sites into five broad categories: 1) peatlands, 2) calcifugous, 3) mesotrophic, 4) dry river sediments and 5) damp river sediments. Further analysis identified 15 ground beetle assemblages, each corresponding to a relatively well defined vegetation type within one of these broad categories. The major environmental factors appearing to determine the distribution of ground beetle assemblages were substratum type, disturbance and soil moisture, all of which are also important determinants of the distribution of plant communities. The presence and absence of relatively stenotopic species were important discriminants of certain habitats such as wetlands and river sediments but the relative abundances within assemblages of more eurytopic species provided good indications of a relationship between ground beetle assemblages and NVC categories.
Foraging behavior of three Tasmanian macropodid marsupials in response to present and historical predation threat
Authors: Blumstein Daniel T. , Daniel Janice C.
Published: October 2003

It is often essential to understand historical selection regimes to explain current traits. We studied antipredator behavior of three Tasmanian macropodid marsupials – Forester kangaroos Macropus giganteus, Bennett's wallabies M. rufogriseus, and Tasmanian pademelons Thylogale billardierii– to understand how antipredator behavior functions in a relatively intact predator community. We also compared behavior of the kangaroos and wallabies on a predator‐free island where they were translocated from mainland Tasmania 30 yr ago. Both species allowed humans to get closer to them on the predator‐free island; a finding consistent with a reduced risk of predation on the island. Neither kangaroos, nor wallabies, exhibited group size effects – they did not modify time allocated to foraging or antipredator vigilance as a function of group size at either site. Nor did overall time allocation vary in any consistent way. In contrast, mainland Australian sibling‐species of Forester kangaroos and Bennett's wallabies have both been reported to have group size effects. It is possible either that the extinction of the thylacine Thylacinus cynocephalus in the last century has led to an evolutionary loss of group size effects and other antipredator behavior, or that thylacines were never that important a predator on Tasmanian subspecies. In contrast, Tasmanian pademelons studied on the Tasmanian mainland modified time allocation as a function of group size suggesting that they perceived safety in numbers. Pademelons, because of their body size, are relatively more vulnerable than larger‐bodied macropodids to the rich community of marsupial carnivores in Tasmania, and used a mix of social and individual strategies to manage predation risk.
The relationships between habitat topology, critical scales of connectivity and tick abundanceIxodes ricinusin a heterogeneous landscape in northern Spain
Authors: Estrada‐Peña Agustin
Published: October 2003

The habitat mosaic was used to quantify connectivity between patches of different tick density of the notorious tick speciesIxodes ricinusin an attempt to determine the cause of variations in tick abundance among apparently homogeneous sites in northern Spain. The analysis revealed that patches with high tick abundance are “stepping‐stone” territories that, when removed from the landscape, cause large changes in connectivity. Sites with medium tick abundance do not cause such a critical transition in connectivity. Patches with low tick abundance, but optimal abiotic conditions for survival, are located within the minimum cost corridors network joining the patches, while those sites where the tick has been intermittently collected are located at variable distances from this network. Sites where the tick is consistently absent, but where the habitat is predicted to be suitable (old, heterogeneous forests ofQuercusspp.) for the tick, are very separated from this main network of connections. These results suggest that tick distribution in a zone is highly affected not only by abiotic variables (vegetation and weather) but also by host movements. Dispersal of the tick is a function of how the hosts perceive the habitat, and the habitat's permeability to host movement. Permanent tick populations seem to be supported by the existence of these critical, high density patches, located at significant places within the habitat network.
Knowledge‐based models for predicting species occurrence in arable conditions
Authors: Petit Sandrine , Chamberlain Dan , Haysom Karen , Pywell Richard , Vickery Juliet , Warman Liz , Allen David , Firbank Les
Published: October 2003

This paper explores the potential of rule‐based habitat models to predict the occurrence of some common species in arable conditions. Models were developed for 10 arable plant species, 7 Hemiptera species, 8 carabid species and for 5 bird species whose ecology was sufficiently known. Rule sets linking species occurrence to environmental variables were produced using available literature and expert knowledge about ecological requirements of the selected species. Environmental variables described the nature and condition of habitats at various scales, ranging from vegetation quadrat to the landscape in a 1 km radius of species sampling sites. The performance of the 34 models developed was assessed in two areas of England. Results show the rule‐based habitat models developed for arable plants and birds were not very successful with Cohen's k values often <0.4 for plants and very close to 0 for all bird species. Conversely, rule‐based models performed surprisingly well for carabids and Hemiptera with k values on average >0.4. This suggests that ecological knowledge on these invertebrate species is more complete than we expected. The effect of species prevalence on model performance and the potential application of knowledge‐based habitat models in the context of biodiversity assessment are discussed.
Migration of the clouded Apollo butterflyParnassius mnemosynein a network of suitable habitats – effects of patch characteristics
Authors: Välimäki Panu , Itämies Juhani
Published: October 2003

The clouded ApolloParnassius mnemosyneis a food plant specialist with short but frequent movements between habitat patches. The short average dispersal distances suggest that the probability of colonisation of vacant patches decreases rapidly as the distance between the source and target patches increases, which means that a dense habitat network is needed for the conservation of the species. Both emigration rate and the number of immigrants varied among patches and were not affected only by isolation but also by several other patch characteristics. The model that explained most of the variation in emigration rates among patches included patch area and the number of conspecifics. The area and the population density of the target patch had significant effects on the number of arriving immigrants. Thus, the colonisation of vacant patches is dependent on these patch characteristics. Generally, emigration rates were lower and residence times longer in large patches with many conspecifics. Butterfly density was the most important single factor explaining the variation in the number of immigrants among patches, although the positive effect of the area of the target patch was also significant. As a consequence of the marked positive density dependence caused by conspecific attraction, small patches with higher than average butterfly density, receive more immigrants than could be expected based on the patch area only. Due to conspecific attraction, per capita immigration rates are higher in small than large patches. Thus, immigration may have a more significant effect on the local dynamics of small than large populations.
Spatial patterns in long‐distance dispersal ofQuercus ilexacorns by jays in a heterogeneous landscape
Authors: Gómez José María
Published: October 2003

In this paper, I analyse the interaction between the holm‐oakQuercus ilex, and one of its main dispersers, the European jayGarrulus glandarius, in an heterogeneous Mediterranean landscape. I quantify the spatial dispersal pattern of the seed shadow at two spatial scales, landscape (among patches) and microhabitat (within patches), by directly tracking the movement of seeds. Two main traits of the jay‐mediated dispersal of holm‐oak acorns across the landscape, the spatial pattern of dissemination and the distance from the source tree, are significantly and directly influenced by jay activity. Jays moved acorns nonrandomly, avoiding one main patch type of the study area to cache acorns, the shrubland‐grasslands, and moving most of the acorns to pine stands, whether afforestation or open pinewoods. Within each patch type, jays had also a strong preference for caching acorns in some microhabitats, since>95% of the acorns dispersed by jays were cached beneath pines. The distance of holm‐oak acorn dispersal was long in the study site, over 250 m, with some dispersals occurring up to 1 km from the source oaks. The shape of the dispersal kernel function fitted to the dispersal pattern produced by jays differed from those quantified for many other plant species. Jay‐mediated dispersal had two components, one local and another produced by long‐distance dispersal. Due to the heterogeneity of these Mediterranean environments, this difference in scale overlaps with a difference in habitat composition, short distances events resulting in dispersals within the same oak stands and long distance events resulting in dispersal outside of oak stands, usually to other vegetation units. Jay activity and movement pattern can have thus dramatic effects on both the local regeneration as well as the potential for regional spread of the holm‐oak populations.
Climatic response of budburst in the mountain birch at two areas in northern Fennoscandia and possible responses to global change
Authors: Karlsson P. S. , Bylund H. , Neuvonen S. , Heino S. , Tjus M.
Published: October 2003

The relationship between the climate and budburst of the mountain birch was evaluated for two areas in subarctic (ca 69°N) Fennoscandia; at Abisko, Swedish Lapland, and at Kevo, Finnish Lapland. Thermal time (TT, degree‐day sums) to budburst was calculated for experimental conditions in the laboratory and for in situ observations of budburst. Two types of models predicting leaf emergence in situ were used: 1) TT to budburst for different threshold temperatures based on daily mean (TTMean) or daily maximum (TTMax) temperatures and 2) ecophysiological budburst models. The obtained models were used to estimate effects of a changed climate.

 Laboratory experiments of TT to budburst indicated no differences in the thermal requirements at the two areas. Temperature requirements of budburst declined successively during the progression of spring, from ca 250 degree‐days (>+2°C) in January to ca 100 in May. No significant trend in the date of budburst was found over the last 70 (Abisko) or 20 (Kevo) yr. There were some differences in the type of model that best explained the date of budburst in situ at the two areas. For Kevo the best prediction (minimum root mean square error, RMSE) of budburst was obtained by a simple thermal time model (TTMean>5.5°C) from 1 January (RMSE=2.1). For budburst at Abisko, models based on daily maximum temperature fitted better than those based on daily means. For Abisko, models based on thermal time accumulation only showed systematic errors in the predicted budburst that were correlated with budburst previous year (BBPY). Including this apparent memory effect in the model decreased the error by 2.4 d. The best prediction for Abisko was thus obtained using TTMAX>6.5 (RMSE=3.1) from 1 January.  Using these models to predict the effect of a changed air temperature climate indicate 3–8 d earlier budburst for a one‐degree increase in temperature, the effect being smaller for Kevo than for Abisko. For both areas a change in May temperature has a larger effect on the date of budburst than changes in any other month.
Effects of anoxia on megakaryocyte progenitors derived from cord blood CD34pos cells
Authors: Saxonhouse Matthew A , Rimsza Lisa M , Christensen Robert D , Hutson Alan D , Stegner Joseph , Koenig Joyce M , Sola Martha C
Published: October 2003

Abstract: Background: Severe hypoxic insults to the fetus and neonate are associated with the development of thrombocytopenia. The thrombocytopenia in some cases is the result of disseminated intravascular coagulation, but that mechanism fails to account for all, perhaps the majority, of cases.

Objective: We hypothesized that human fetal megakaryocyte (Mk) progenitors are directly adversely affected by transient anoxia.

Design and methods: To test this, we isolated CD34pos cells from the umbilical cord blood of 10 healthy term neonates, and exposed these to 0% or 20% O2 for 24 h, with or without recombinant thrombopoietin (rTpo, 50 ng/mL). After 24 h, a portion of the CD34pos cells were harvested for flow cytometric evaluation of apoptosis. The remaining cells were cultured for an additional 10–12 days, under normoxic conditions, in a collagen‐based serum‐free system containing rTpo, IL‐3, and IL‐6. In this way, we sought to determine the effect of transient anoxia on clonogenic capacity of Mk progenitors.

Results: Contrary to our hypothesis, anoxia did not increase either apoptosis or cell death of the CD34pos cells. The addition of rTpo was protective, with a significant decrease in apoptosis and cell death (P < 0.0001), and an increase in the number of Mk colonies cultured (P = 0.04). There was no difference between the normoxic and anoxic groups in proliferative potential of the Mk progenitor cells.

Conclusions: The thrombocytopenia observed in neonates following an acute hypoxic event is not likely due to a direct deleterious effect of hypoxia on Mk progenitors.
Elevated plasma levels of TIMP‐1 correlate with plasma suPAR/uPA in patients with chronic myeloproliferative disorders
Authors: Jensen Morten Krogh , Holten‐Andersen Mads Nikolaj , Riisbro Rikke , de Nully Brown Peter , Larsen Maj Britt , Kjeldsen Lars , Heickendorff Lene , Brünner Nils , Hasselbalch Hans Carl
Published: October 2003

Abstract: Chronic myeloproliferative disorders (MPD) are characterized by progressive remodelling of bone marrow stroma as evidenced by increased deposition of extracellular matrix proteins, neoangiogenesis and displacement of normal haematopoietic cells by fibrotic tissue. The family of metalloproteinases (MMPs) and tissue inhibitors of metalloproteinase (TIMPs) serve to facilitate and inhibit matrix degradation processes, respectively. In an attempt to investigate potential markers for bone marrow remodelling processes, we investigated plasma levels of total‐, free‐ and complexed TIMP‐1, TIMP‐2, MMP‐2 and MMP‐9 in a patient cohort comprising 17 with myelofibrosis (MF), 17 with polycythaemia vera (PV), 15 with essential thrombocythaemia (ET), 1 with a transitional MPD and 30 controls. Compared with controls, total‐ (P < 0.0001) (median: 132.6 μg/L vs. 80.8 μg/L), free‐ (P < 0.0001) (median: 126.4 μg/L vs. 65.8 μg/L) and complexed TIMP‐1 (P = 0.0009) (median: 17.7 μg/L vs. 10.7 μg/L) concentration was significantly higher in the patients. TIMP‐1 was significantly correlated with plasma soluble urokinase plasminogen activator receptor (P = 0.003) and urokinase plasminogen activator (P < 0.0001), respectively, suggesting a common cellular origin. No statistical significant difference between TIMP‐2 and MMP‐2 levels was observed between patients and controls. Furthermore, a significant correlation between free TIMP‐1 and TIMP‐2 levels was detected (r = 0.56; P < 0.0001). Median MMP‐9 concentration was significantly higher among PV patients compared with controls (P = 0.0015), and 41% of patients with PV (7/17) had MMP‐9 values that were above the mean + 2SD of plasma MMP‐9 levels found in controls. The ratio of total TIMP‐1/MMP‐9 was significantly higher in patients with MF compared with controls (P = 0.0004). These findings suggest that a disturbed TIMP‐1/MMP ratio may reflect an imbalance of the extracellular homeostasis towards an increased matrix deposition promoting fibrosis.
Aplastic anemia following administration of a tumor necrosis factor‐α inhibitor
Authors: Kuruvilla John , Leitch Heather A. , Vickars Linda M. , Galbraith Paul F. , Li Charles H. , Al‐Saab Saad , Naiman Sheldon C.
Published: October 2003

Abstract: Upregulation of tumor necrosis factor‐alpha (TNF‐α) has been implicated in the pathogenesis of several inflammatory conditions, including rheumatoid arthritis. Therapeutic agents such as antibodies or soluble TNF‐α receptor analogs, which block TNF‐α activity are a recent addition to the therapeutic armamentarium for the conditions. We describe a patient who developed aplastic anemia complicated by sepsis after receiving etanercept, a TNF‐α receptor analog, for the treatment of rheumatoid arthritis. Pancytopenia resolved within 3 wk of discontinuing etanercept. To our knowledge, this is the first report of aplastic anemia associated with TNF‐α blockade.
A 29‐yr‐old man with multiple myeloma presenting with subcutaneous masses
Authors: Procopiou Michel , Hügli Anne
Published: October 2003

Abstract: We report the case of a 29‐yr‐old man with multiple myeloma who presented initially with subcutaneous plasmacytomas. These are the atypical features of his clinical course: (a) age of the patient; (b) unusual clinical presentation with subcutaneous masses; (c) lack of ‘typical’ features of multiple myeloma (renal failure, hypercalcaemia, and bone pain); and (d) disease progression as extramedullary recurrences.
Multiparametric analysis of immature cell populations in umbilical cord blood and bone marrow
Authors: McGuckin Colin P. , Pearce Daniel , Forraz Nicolas , Tooze Jennifer A. , Watt Suzanne M. , Pettengell Ruth
Published: October 2003

Abstract: Adult stem cells are finding increased therapeutic potential not least in tissue regeneration protocols. The cell sources being proposed for such protocols include embryonic, umbilical cord blood (CB) and adult bone marrow (BM). Although embryonic sources are controversial, CB and marrow are available immediately. The appropriate cells of use in these sources are considered to be extremely rare and a characterisation of the starting cell source is important for the development of adult stem cell protocols and ex vivo expansion. Umbilical CB and BM mononuclear cells were labelled for the antigens CD34, CD133, CD117, CD164, Thy‐1 or CD38, and additional intracellular CD34 antigen. Three dimensional flow‐cytometric analyses were carried out together with dual laser confocal microscope analysis for antigen profile expression. Variable levels of immaturity were detected on CB and BM populations using internal and external CD34 antigen. For CB and BM cells, internal CD34 (intCD34+) could be detected on co‐expressing CD133+ cells before expression of external CD34 antigen (extCD34+). CD38 co‐expression analysis also showed that a small but distinct group of cells expressing low CD38 and no external CD34 antigen could be detected. Additional phenotyping of these cells using CD117, Thy‐1, CD164 and CD133 demonstrated variable primitive status detectable within the external CD34− population. Newly harvested primary CB and BM populations were shown to contain not only cellular populations of known standard sequential maturity but also populations of more extreme rarity. The presence of cells which lacked extracellular CD34 antigen, in both CB and BM, but which possessed CD133, has important implications for purification of human stem cells in clinical applications.
Prevalence of oral mucosal lesions in elderly people in Santiago, Chile
Authors: Espinoza I. , Rojas R. , Aranda W. , Gamonal J.
Published: October 2003

Background:  Oral prevalence studies are important to know the state of health and the needs of treatment. Our aim was to determine the prevalence of oral mucosal lesions and associated factors among aging Chileans.

Methods:  A random sample by age, gender, and socioeconomic status was obtained, comprising 889 individuals older than 65 years. Individuals were interviewed and examined in Santiago, the capital of Chile, according to the World Health Organization guidelines.

Results:  The prevalence of one or more oral mucosal lesions in the sample was 53%. Logistic regression model revealed that denture use increased the probability of one or more oral mucosal lesions by threefold, while age, gender, smoking, medication use, xerostomia, and social or cultural factors had no effect. The most common lesion was denture stomatitis (22.3%), followed by irritative hyperplasia (9.4%), oral mucosal varicosities (9%), solitary pigmented lesions (4%), traumatic ulcer (3.5%), angular cheilitis (2.9%), multiple pigmented lesions (2.8%), hemangioma (2.3%), lichen planus (2.1%), leukoplakia (1.7%), recurrent aphthous stomatitis (1.4%), nicotine stomatitis (1.3%), median rhomboid glossitis (0.9%), actinic cheilitis (0.9%), pyogenic granuloma (0.7%), oral squamous papiloma (0.6%), and mucocele (0.2%). One case of oral cancer was observed. Different factors increased the probability of specific oral mucosal pathologies.

Conclusions:  We can conclude that oral mucosal lesions are common in elderly people in Santiago, suggesting the necessity for improved standards of prevention, and diagnostic and opportune treatment of these lesions.
Orofacial granulomatosis: presentation, pathology and management of 13 cases
Authors: Sciubba James J. , Said‐Al‐Naief Nasser
Published: October 2003

Background: Orofacial granulomatosis (OFG) comprises of a group of conditions, all characterized histologically by the presence of granulomatous inflammation.

Methods: This diagnosis may be determined by exclusion from other conditions that may present with similar clinical and histopathologic features. These include Melkersson–Rosenthal syndrome (MRS), Miescher's cheilitis (an oligosymptomatic form of MRS), Crohn's disease, and sarcoidosis.

Results: A great deal of attention has been devoted to the similarity and overlap in clinicopathologic and histomorphologic features of these conditions, suggesting that they may actually represent a spectrum within a single overarching entity. In the review of the 13 cases of OFG retrieved from the files of the Long Island Jewish Medical Center, Department of Dental Medicine, we describe their presentation, clinicopathologic features, and management. These cases comprise examples of MRS (in its oligosymptomatic forms) and Crohn's disease. The similarity, kinship, and overlap between the cases presented are clearly demonstrated.

Conclusion: In addition, based upon our observations and review, we propose the notion that oral manifestations of Crohn's disease may be classified as an oligosymptomatic form of MRS.
Insulin Signaling in Microdomains of the Plasma Membrane
Authors: Saltiel Alan R. , Pessin Jeffrey E.
Published: October 2003

Although the effects of insulin on glucose and lipid metabolism are well documented, gaps remain in our understanding of the precise molecular mechanisms of signal transduction. Recent evidence suggests that compartmentalization of signaling molecules and metabolic enzymes may explain the unique cellular effects of the hormone. Signal initiation from the insulin receptor is restricted in part to caveolae microdomains of the plasma membrane. A fraction of the insulin receptor directly interacts with caveolin, thus directing the protein to caveolae. Following its activation by insulin, the receptor recruits a series of adapter proteins, resulting in the activation of the G protein TC10, which also resides in caveolae. TC10 can influence a number of cellular processes, including changes in the actin cytoskeleton, recruitment of effector including the adapter protein CIP4, and assembly of the exocyst complex. These events play crucial roles in the trafficking, docking and fusion of vesicles containing the insulin‐responsive glucose transporter Glut4 at the plasma membrane.
Lipid Rafts and Caveolae as Portals for Endocytosis: New Insights and Common Mechanisms
Authors: Parton Robert G. , Richards Ayanthi A.
Published: October 2003

Clathrin‐coated pits and caveolae are two of the most recognizable features of the plasma membrane of mammalian cells. While our understanding of the machinery regulating and driving clathrin‐coated pit‐mediated endocytosis has progressed dramatically, including the elucidation of the structure of individual components and partial in vitro reconstitution, the role of caveolae as alternative endocytic carriers still remains elusive 50 years after their discovery. However, recent work has started to provide new insights into endocytosis by caveolae and into apparently related pathways involving lipid raft domains. These pathways, distinguished by their exquisite sensitivity to cholesterol‐sequestering agents, can involve caveolae but also exist in cells devoid of caveolins and caveolae. This review examines the current evidence for the involvement of rafts and caveolae in endocytosis and the molecular players involved in their regulation.
Human VPS34 and p150 are Rab7 Interacting Partners
Authors: Stein Mary‐Pat , Feng Yan , Cooper Karen L. , Welford Angela M. , Wandinger‐Ness Angela
Published: October 2003

Regulation of membrane trafficking requires the concerted actions of rab proteins, their effectors and several phosphatidylinositol 3′‐kinases. Rab7 is required for late endosomal transport and here we establish that the phosphatidylinositol 3′‐kinase hVPS34 and its adaptor protein p150 are rab7 interacting partners. The hVPS34/p150 complex colocalized with rab7 on late endosomes and hVPS34 activity was dependent on nucleotide cycling of rab7. In addition, total cellular phosphatidylinositol 3′‐phosphate levels were modulated by rab7 expression, suggesting that rab7 activation impacted kinase cycling to early endosomes. The data identify rab7 as an important regulator of late endosomal hVPS34 function and link rab7 to the regulation of phosphatidylinositol 3′‐kinase cycling between early and late endosomes.
Visualization of Retroviral Replication in Living Cells Reveals Budding into Multivesicular Bodies
Authors: Sherer Nathan M. , Lehmann Maik J. , Jimenez‐Soto Luisa F. , Ingmundson Alyssa , Horner Stacy M. , Cicchetti Gregor , Allen Philip G. , Pypaert Marc , Cunningham James M. , Mothes Walther
Published: October 2003

Retroviral assembly and budding is driven by the Gag polyprotein and requires the host‐derived vacuolar protein sorting (vps) machinery. With the exception of human immunodeficiency virus (HIV)‐infected macrophages, current models predict that the vps machinery is recruited by Gag to viral budding sites at the cell surface. However, here we demonstrate that HIV Gag and murine leukemia virus (MLV) Gag also drive assembly intracellularly in cell types including 293 and HeLa cells, previously believed to exclusively support budding from the plasma membrane. Using live confocal microscopy in conjunction with electron microscopy of cells generating fluorescently labeled virions or virus‐like particles, we observed that these retroviruses utilize late endosomal membranes/multivesicular bodies as assembly sites, implying an endosome‐based pathway for viral egress. These data suggest that retroviruses can interact with the vps sorting machinery in a more traditional sense, directly linked to the mechanism by which cellular proteins are sorted into multivesicular endosomes.
Bioinformatic analysis of autism positional candidate genes using biological databases and computational gene network prediction
Authors: Yonan A. L. , Palmer A. A. , Smith K. C. , Feldman I. , Lee H. K. , Yonan J. M. , Fischer S. G. , Pavlidis P. , Gilliam T. C.
Published: October 2003

Common genetic disorders are believed to arise from the combined effects of multiple inherited genetic variants acting in concert with environmental factors, such that any given DNA sequence variant may have only a marginal effect on disease outcome. As a consequence, the correlation between disease status and any given DNA marker allele in a genomewide linkage study tends to be relatively weak and the implicated regions typically encompass hundreds of positional candidate genes. Therefore, new strategies are needed to parse relatively large sets of ‘positional’ candidate genes in search of actual disease‐related gene variants. Here we use biological databases to identify 383 positional candidate genes predicted by genomewide genetic linkage analysis of a large set of families, each with two or more members diagnosed with autism, or autism spectrum disorder (ASD). Next, we seek to identify a subset of biologically meaningful, high priority candidates. The strategy is to select autism candidate genes based on prior genetic evidence from the allelic association literature to query the known transcripts within the 1‐LOD (logarithm of the odds) support interval for each region. We use recently developed bioinformatic programs that automatically search the biological literature to predict pathways of interacting genes (pathwayassist and geneways). To identify gene regulatory networks, we search for coexpression between candidate genes and positional candidates. The studies are intended both to inform studies of autism, and to illustrate and explore the increasing potential of bioinformatic approaches as a compliment to linkage analysis.
The amygdala and autism: implications from non‐human primate studies
Authors: Amaral D. G. , Bauman M. D. , Mills Schumann C.
Published: October 2003

Brothers (1990) has proposed that the amygdala is an important component of the neural network that underlies social behavior.Kemper and Bauman (1993) identified neuropathology in the amygdala of the postmortem autistic brain. These findings, along with recent functional neuroimaging data, have ledBaron‐Cohen et al. (2000) to propose that dysfunction of the amygdala may be responsible, in part, for the impairment of social behavior that is a hallmark feature of autism. Recent data from studies in our laboratory on the effects of amygdala lesions in the adult and infant macaque monkey do not support a fundamental role for the amygdala in social behavior. If the amygdala is not essential for the component processes of social behavior, as seems to be case in both non‐human primates and selected patients with bilateral amygdala damage, then it is unlikely to be the primary substrate for the impaired social behavior of autism. However, damage to the amygdala does have an effect on a monkey's response to normally fear‐inducing stimuli, such as snakes, and removes a natural reluctance to engage novel conspecifics in social interactions. These findings lead to the conclusion that an important role for the amygdala is in the detection of threats and mobilizing an appropriate behavioral response, part of which is fear. Interestingly, an important comorbid feature of autism is anxiety (Muris et al. 1998). If the amygdala is pathological in subjects with autism, it may contribute to their abnormal fears and increased anxiety rather than their abnormal social behavior.
How to select the right mouthrinses in periodontal prevention and therapy. Part II. Clinical use and recommendations
Authors: Brecx M , Netuschil L , Hoffmann T
Published: October 2003

Abstract:  Based on scientific proof, only few mouthrinse solutions can be recommended. However, it is impossible to construct an extended table that would provide precise recommendations or advices for mouthrinse utilisation related to any possible discomfort or diseases. Only reliable information on the drugs available against periodontal disease can be offered to the therapist. By evaluation of her or his individual experiences, she or he has to judge all the armamentarium she or he possesses to react in the best possible way in each specific situation.
Social relations and breath odour
Authors: McKeown L
Published: October 2003

Abstract:  In this retrospective qualitative study, the researcher reviewed 55 client records of The Breath Odour Clinic. The purpose was to determine if individuals attended a clinic specialised in treating oral malodour for medical or social reasons. The study focused on the psychosocial and breath odour history. Clients had agreed to the use of information for research purposes. Society uses odour as a means to define and interact with the world. The olfactory, smelling experience is intimate, emotionally charged and connects us with the world. It follows that the smell from mouth breath odour can connect or disconnect a person from their social environment and intimate relationships. How one experiences one's own body is very personal and private but also very public. Breath odour is public as it occurs within a social and cultural context and personal as it affects one's body image and self‐confidence. Body image, self‐image and social relations mesh, interact and impact upon each other. Breath odour is a dynamic and interactive aspect of the self‐image. In addition, breath odour may be value‐coded as ‘bad’. In 75% of the cases reviewed, decreased self‐confidence and insecurity in social and intimate relations led clients to seek treatment at the specialised breath odour clinic. Their doctor, dental hygienist or dentist had treated medical and oral conditions but not resolved their breath odour problem. When a person perceives a constant bad breath problem, she/he uses defence techniques, and may avoid social situations and social relations. This affects a person's well‐being.
The unique features of glycolytic pathways in Archaea
Authors: VERHEES Corné H. , KENGEN Servé W. M. , TUININGA Judith E. , SCHUT Gerrit J. , ADAMS Michael W. W. , de VOS Willem M. , OOST John van der
Published: October 2003

An early divergence in evolution has resulted in two prokaryotic domains, the Bacteria and the Archaea. Whereas the central metabolic routes of bacteria and eukaryotes are generally well-conserved, variant pathways have developed in Archaea involving several novel enzymes with a distinct control. A spectacular example of convergent evolution concerns the glucose-degrading pathways of saccharolytic archaea. The identification, characterization and comparison of the glycolytic enzymes of a variety of phylogenetic lineages have revealed a mosaic of canonical and novel enzymes in the archaeal variants of the Embden–Meyerhof and the Entner–Doudoroff pathways. By means of integrating results from biochemical and genetic studies with recently obtained comparative and functional genomics data, the structure and function of the archaeal glycolytic routes, the participating enzymes and their regulation are re-evaluated.
Forkhead transcription factor FOXO1 (FKHR)-dependent induction of PDK4 gene expression in skeletal muscle during energy deprivation
Authors: FURUYAMA Tatsuo , KITAYAMA Kazuko , YAMASHITA Hitoshi , MORI Nozomu
Published: October 2003

A forkhead-type transcription factor, DAF-16, is located in the most downstream part of the insulin signalling pathway via PI3K (phosphoinositide 3-kinase). It is essential for the extension of life-span and is also involved in dauer formation induced by food deprivation in Caenorhabditis elegans. In the present study, we addressed whether or not FOXO members AFX, FKHR (forkhead homologue in rhabdomyosarcoma) and FKHRL1 (FKHR-like protein 1), mammalian counterparts of DAF-16, are involved in starvation stress. We found a remarkable selective induction of FKHR and FKHRL1 transcripts in skeletal muscle of mice during starvation. The induction of FKHR gene expression was observed at 6 h after food deprivation, peaked at 12 h, and returned to the basal level by 24 h of refeeding. The induction was also found in skeletal muscle of mice with glucocorticoid treatment. Moreover, we found that the levels of PDK4 (pyruvate dehydrogenase kinase 4) gene expression were up-regulated through the direct binding of FKHR to the promoter region of the gene in C2C12 cells. These results suggest that FKHR has an important role in the regulation of energy metabolism, at least in part, through the up-regulation of PDK4 gene expression in skeletal muscle during starvation.
The conserved cis-Pro39 residue plays a crucial role in the proper positioning of the catalytic base Asp38 in ketosteroid isomerase from Comamonas testosteroni
Authors: NAM Gyu Hyun , CHA Sun-Shin , YUN Young Sung , OH Yun Hee , HONG Bee Hak , LEE Heung-Soo , CHOI Kwan Yong
Published: October 2003

KSI (ketosteroid isomerase) from Comamonas testosteroni is a homodimeric enzyme that catalyses the allylic isomerization of Δ5-3-ketosteroids to their conjugated Δ4-isomers at a reaction rate equivalent to the diffusion-controlled limit. Based on the structural analysis of KSI at a high resolution, the conserved cis-Pro39 residue was proposed to be involved in the proper positioning of Asp38, a critical catalytic residue, since the residue was found not only to be structurally associated with Asp38, but also to confer a structural rigidity on the local active-site geometry consisting of Asp38, Pro39, Val40, Gly41 and Ser42 at the flexible loop between β-strands B1 and B2. In order to investigate the structural role of the conserved cis-Pro39 residue near the active site of KSI, Pro39 was replaced with alanine or glycine. The free energy of activation for the P39A and P39G mutants increased by 10.5 and 16.7 kJ/mol (2.5 and 4.0 kcal/mol) respectively, while ΔGUH2O (the free-energy change for unfolding in the absence of urea at 25.00±0.02 °C) decreased by 31.0 and 35.6 kJ/mol (7.4 and 8.5 kcal/mol) respectively, compared with the wild-type enzyme. The crystal structure of the P39A mutant in complex with d-equilenin [d-1,3,5(10),6,8-estrapentaen-3-ol-17-one], a reaction intermediate analogue, determined at 2.3 Å (0.23 nm) resolution revealed that the P39A mutation significantly disrupted the proper orientations of both d-equilenin and Asp38, as well as the local active-site geometry near Asp38, which resulted in substantial decreases in the activity and stability of KSI. Upon binding 1-anilinonaphthalene-8-sulphonic acid, the fluorescence intensities of the P39A and P39G mutants were increased drastically, with maximum wavelengths blue-shifted upon binding, indicating that the mutations might alter the hydrophobic active site of KSI. Taken together, our results demonstrate that the conserved cis-Pro39 residue plays a crucial role in the proper positioning of the critical catalytic base Asp38 and in the structural integrity of the active site in KSI.
Inhibition of human ornithine decarboxylase activity by enantiomers of difluoromethylornithine
Authors: QU Ning , IGNATENKO Natalia A. , YAMAUCHI Phillip , STRINGER David E. , LEVENSON Corey , SHANNON Patrick , PERRIN Scott , GERNER Eugene W.
Published: October 2003

Racemic difluoromethylornithine (d/l-DFMO) is an inhibitor of ODC (ornithine decarboxylase), the first enzyme in eukaryotic polyamine biosynthesis. d/l-DFMO is an effective anti-parasitic agent and inhibitor of mammalian cell growth and development. Purified human ODC-catalysed ornithine decarboxylation is highly stereospecific. However, both DFMO enantiomers suppressed ODC activity in a time- and concentration-dependent manner. ODC activity failed to recover after treatment with either l- or d-DFMO and dialysis to remove free inhibitor. The inhibitor dissociation constant (KD) values for the formation of enzyme–inhibitor complexes were 28.3±3.4, 1.3±0.3 and 2.2±0.4 μM respectively for d-, l- and d/l-DFMO. The differences in these KD values were statistically significant (P<0.05). The inhibitor inactivation constants (Kinact) for the irreversible step were 0.25±0.03, 0.15±0.03 and 0.15±0.03 min−1 respectively for d-, l- and d/l-DFMO. These latter values were not statistically significantly different (P>0.1). d-DFMO was a more potent inhibitor (IC50~7.5 μM) when compared with d-ornithine (IC50~1.5 mM) of ODC-catalysed l-ornithine decarboxylation. Treatment of human colon tumour-derived HCT116 cells with either l- or d-DFMO decreased the cellular polyamine contents in a concentration-dependent manner. These results show that both enantiomers of DFMO irreversibly inactivate ODC and suggest that this inactivation occurs by a common mechanism. Both enantiomers form enzyme–inhibitor complexes with ODC, but the probability of formation of these complexes is 20 times greater for l-DFMO when compared with d-DFMO. The rate of the irreversible reaction in ODC inactivation is similar for the l- and d-enantiomer. This unexpected similarity between DFMO enantiomers, in contrast with the high degree of stereospecificity of the substrate ornithine, appears to be due to the α-substituent of the inhibitor. The d-enantiomer may have advantages, such as decreased normal tissue toxicity, over l- or d/l-DFMO in some clinical applications.
Identification and characterization of Snapin as a ubiquitously expressed SNARE-binding protein that interacts with SNAP23 in non-neuronal cells
Authors: BUXTON Penelope , ZHANG Xiang-Ming , WALSH Bong , SRIRATANA Absorn , SCHENBERG Irina , MANICKAM Elizabeth , ROWE Tony
Published: October 2003

Members of the SNARE (soluble N-ethylmaleimide-sensitive fusion protein attachment protein receptor) superfamily [syntaxins, VAMPs (vesicle-associated membrane proteins) and SNAP25 (synaptosome-associated protein-25)-related proteins] are required for intracellular membrane-fusion events in eukaryotes. In neurons, assembly of SNARE core complexes comprising the presynaptic membrane-associated SNAREs syntaxin 1 and SNAP25, and the vesicle-associated SNARE VAMP2, is necessary for synaptic vesicle exocytosis. Several accessory factors have been described that associate with the synaptic SNAREs and modulate core complex assembly or mediate Ca2+ regulation. One such factor, Snapin, has been reported to be a brain-specific protein that interacts with SNAP25, and regulates association of the putative Ca2+-sensor synaptotagmin with the synaptic SNARE complex [Ilardi, Mochida and Sheng (1999) Nat. Neurosci. 2, 119–124]. Here we demonstrate that Snapin is expressed ubiquitously in neuronal and non-neuronal cells. Furthermore, using protein–protein-interaction assays we show that Snapin interacts with SNAP23, the widely expressed homologue of SNAP25, and that the predicted C-terminal helical domain of Snapin contains the SNAP23-binding site. Subcellular localization experiments revealed that Snapin is a soluble protein that exists in both cytosolic and peripheral membrane-bound pools in adipocytes. Moreover, association of Snapin with the plasma membrane was detected in cells overexpressing a Snapin–green fluorescent protein fusion protein. Finally, we show that Snapin is able to form a ternary complex with SNAP23 and syntaxin 4, suggesting that it is a component of non-neuronal SNARE complexes. An important implication of our results is that Snapin is likely to perform a general role in SNARE-mediated vesicle fusion events in non-neuronal cells in addition to its participation in Ca2+-regulated neurosecretion.