bluuebunny/crossref_metadata_2025 huggingface dataset:
FOXP3 acts as a rheostat of the immune response
Authors: Ochs Hans D. , Ziegler Steven F. , Torgerson Troy R.
Published: January 2005

Summary:  The study of a rare human X‐linked disease resulting in a characteristic clinical phenotype of multiple autoimmune disorders and the in‐depth exploration of a spontaneous mouse model, scurfy (sf), have contributed to a better understanding of the regulation of immunologic responses, particularly to self. Forkhead box P3 (FOXP3), the gene responsible for IPEX (immune dysregulation, polyendocrinopathy, enteropathy, X‐linked) and sf is located on the X chromosome and is of crucial importance for the generation of CD4+CD25+ regulatory T cells. Loss of FOXP3 function and the resultant lack of regulatory T cells result in lethal auto‐aggressive lymphoproliferation, whereas overexpression of this modulator results in severe immunodeficiency. The in‐depth analysis of FOXP3 regulation and elucidation of the precise mechanisms by which FOXP3 exerts its regulatory effect will provide important insights into the understanding of autoimmunity and should predictably result in new therapeutic possibilities.
Interleukin‐7 receptor α (IL‐7Rα) deficiency: cellular and molecular bases. Analysis of clinical, immunological, and molecular features in 16 novel patients
Authors: Giliani Silvia , Mori Luigi , De Saint Basile Geneviève , Le Deist Francoise , Rodriguez‐Perez Carmen , Forino Concetta , Mazzolari Evelina , Dupuis Sophie , Elhasid Ronit , Kessel Aharon , Galambrun Claire , Gil Juana , Fischer Alain , Etzioni Amos , Notarangelo Luigi D.
Published: January 2005

Summary:  Analysis of gene‐targeted mice and patients with severe combined immunodeficiency due to mutations of the α chain of the interleukin‐7 receptor (IL‐7Rα) has shown important differences between mice and humans in the role played by IL‐7 in lymphoid development. More recently, it has been shown that IL‐7Rα is also shared by the receptor for another cytokine, thymic stromal lymphopoietin (TSLP). In this review, we discuss recent advances in IL‐7‐ and TSLP‐mediated signaling. We also report on the clinical and immunological features of 16 novel patients with IL‐7Rα deficiency and discuss the results of hematopoietic stem cell transplantation.
BERNINI'S LIGHT
Authors: Fehrenbach Frank
Published: January 2005

Art historians have traditionally offered dualistic interpretations of Bernini's stagings of light, treating the light as the ‘metaphor’, ‘symbol’ or ‘sign’ of an invisible, ‘heavenly’ light. Such interpretations, however, oversimplify a more complex set of relationships. As the biography by Bernini's son Domenico makes clear, the spirit, the bearer of artistic talent or ingenium, is a light‐like imponderable; using this ‘inner’ light, Bernini enlivened his media. Thus, visible light appears as an actor in Bernini's work. His sculptures react to it; they are emphatically and ecstatically moved by it. Contemporaries characterized physical light, paradoxically, as both corporeal and incorporeal, and under Bernini's direction, the two levels of interpretation were combined. Bernini's light manifests his ingenium and, at the same time, the divine that penetrates the perceptible world. Better to comprehend the participation of the physical in the spiritual, this article suggests, one should speak of Bernini's light not in terms of ‘metaphor’, but of ‘analogy’.
Time tells: long‐term patterns in the population dynamics of the yew gall midge,Taxomyia taxi(Cecidomyiidae), over 35 years
Authors: Redfern Margaret , Hunter Mark D.
Published: January 2005

Abstract. 1. Three populations of the yew gall midge,Taxomyia taxi, and its two chalcid parasitoids,Mesopolobus diffinisandTorymus nigritarsus, from Kingley Vale, West Sussex, U.K., were monitored each year between 1966 and 2002.Taxomyia taxi's life cycles are either hemivoltine or univoltine and the parasitoids' are univoltine (T. nigritarsus) or multivoltine (M. diffinis).

2. Growth of the host trees, yewTaxus baccata, each year between 1950 and 1997 was assessed by width of annual rings in cores taken in 1998. Temperature and rainfall data were provided by the weather station in Southampton, representative of the climate of central southern England.

3. Time‐series analyses were conducted to explore the effects of climate, tree growth, and parasitoids on eachT. taxipopulation and on the three populations combined. Four time‐delayed models were developed: for hemivoltine and univoltineT. taxiand for each of the parasitoids.

4. HemivoltineT. taxipopulations showed troughs in density in the periods 1972–1975 and 1991–1992, which were more pronounced in even than odd years; these were closely tracked byM. diffinisand followed, after a delay, byT. nigritarsus. Time‐series models showed that, forT. taxi, its current density depended on the density of its hemivoltine parents moderated by the negative effects ofT. nigritarsuson these parents. In reciprocal fashion,T. nigritarsusdensity was influenced by that of its hemivoltine host as well as by its own previous density. Host and parasitoid appear to participate in a coupled interaction that regulates the populations of both via delayed density dependence, resulting in cycles of at least 14 years.Mesopolobus diffinishad no effect on the dynamics of hemivoltineT. taxi, most of which escaped its parasitism. Although univoltineT. taxican give rise to hemivoltine offspring, there was no evidence that they affected hemivoltine dynamics.

5. The density of univoltineT. taxiwas usually much lower than hemivoltine density and varied irregularly over time. It was not attacked byT. nigritarsusbut parasitism byM. diffiniswas always high. Despite this, its density depended only on the density of its parents, especially the hemivoltine parents. Parasitism had no effect on the dynamics of univoltineT. taxi. Conversely,M. diffinisdensity depended solely on the densities of both univoltine and hemivoltine hosts.

6. Tree growth appeared to have little effect on the dynamics of the host or parasitoids and there was no evidence that gall density had any effect on growth of the tree. The primary effect of climate appeared to be in maintaining the cyclic nature of theT. taxi–T. nigritarsusinteraction. If stochastic weather variables are removed from the models, the oscillations are of equivalent periodicity but dampen to equilibrium.
The relationship between the abundances of bumblebees and honeybees in a native habitat
Authors: Forup Mikael Lytzau , Memmott Jane
Published: January 2005

Abstract.  1. The western honeybee, Apis mellifera, has been introduced to many parts of the world and is sometimes purported to be detrimental to native bees because it reduces their food base. It is seldom viewed in this light in Europe; however, when beekeepers maintain very high bee densities, the species could also be displacing insects in its native European range by reducing the resource base.

2. In England, populations of bumblebees (Bombus Latr. Hym.) have been decreasing both in terms of diversity and abundance, mainly because of a loss of habitat resulting from agricultural intensification. The impact of competition from other flower feeders is largely unknown.

3. Nineteen dry lowland heaths in southern England were sampled once for honeybees and bumblebees. Honeybee abundance varied from 4 to 81 bees per 100 m2 (mean = 30.89, median = 23), whereas bumblebees varied from 2 to 17 individuals per 100 m2 (mean = 8.26, median = 7), belonging to between one and five species. There was a negative association between honeybee and bumblebee abundance but there was no apparent relationship between honeybee abundance and bumblebee diversity.

4. The Bray–Curtis coefficient was used to compare the similarity in honeybee and bumblebee floral host breadth at these 19 sites. The coefficient was negatively associated with honeybee abundance: thus where honeybees were most abundant, bumblebees were fewer and/or foraged on different flower species.

5. Foraging host breadth was also examined at four heathlands over a field season (April to September). No association between honeybee abundance and foraging host breadth was found for short‐tongued bees, although there was some evidence for a change in floral host breadth for long‐tongued bees.

6. It is concluded that the impact of honeybees on bumblebees is complex. Although competition between the two species cannot be ruled out, it is perhaps equally likely that bumblebees decline in response to other factors, and that honeybees move independently of this decline.
Asynchronous temporal variation among sites in condition of two carabid species
Authors: Östman Örjan
Published: January 2005

Abstract.  1. It is generally assumed that generalist predatory carabids are food limited, and the degree of food limitation can vary in space and time, but there are only a few studies that investigate whether or not temporal variation in insect condition is synchronised among sites.

2. In this paper temporal and spatial variation in four fitness‐related traits – body mass corrected for size, body size, fat content, and fecundity – of two carabid species, Pterostichus cupreus (L.) and Pterostichus melanarius (Ill.), differing in overwintering strategy, lifespan, diet, and habitat breadth were studied.

3. Interactions between temporal and spatial variation for the fitness‐related traits were found for both species. Temporal changes in food limitation were not synchronised among sites, and fitness‐related traits were not generally higher on some sites than on others.

4. There was an effect of study year on body mass, fat content, and fecundity for both species, suggesting that temporal environmental variation over large areas is more important than the spatial environmental variation within a year.

5. Within a time period, most different fitness‐related traits, except body size, were positively correlated to each other among individuals of the same species. The fitness‐related traits of females and males of the same species were generally positively correlated. However, despite the fact that the food resources of the two species overlap, there was no correlation in fitness‐related traits between the two species.
Is the matrix a sea? Habitat specificity in a naturally fragmented landscape
Authors: Driscoll Don A.
Published: January 2005

Abstract.  1. Metapopulation and island biogeography theory assume that landscapes consist of habitat patches set in a matrix of non‐habitat. If only a small proportion of species conform to the patch–matrix assumptions then metapopulation theory may only describe special cases rather than being of more general ecological importance.

2. As an initial step towards understanding the prevalence of metapopulation dynamics in a naturally fragmented landscape, the distribution of beetle species in three replicates of three habitat types was examined, including rainforest and eucalypt forest (the habitat patches), and buttongrass sedgeland (the matrix), in south‐west Tasmania, Australia.

3. Ordination methods indicated that the buttongrass fauna was extremely divergent from the fauna of forested habitats. Permutation tests showed that the abundance of 13 of 17 commonly captured species varied significantly among habitats, with eight species confined to eucalypts or rainforest, and three species found only in buttongrass. Approximately 60% of species were confined to forested habitat implying that metapopulation theory has the potential to be very important in the forest–buttongrass landscape.

4. Although floristically the rainforest and eucalypts were extremely distinct, the beetle faunas from eucalypts and rainforests overlapped substantially. Therefore rainforest patches connected by eucalypt forest represent continuous habitat for most species.

5. Other studies report a wide range of values for the proportion of patch‐specific species in fragmented landscapes. Understanding the environmental or historical conditions under which a high proportion of species become patch specialists would help to identify where spatial dynamic theory may be especially applicable, and where habitat loss and fragmentation poses the greatest threat to biodiversity.
Bottom‐up cascade in a tri‐trophic system: different impacts of host‐plant regeneration on performance of a willow leaf beetle and its natural enemy
Authors: Kagata Hideki , Nakamura Masahiro , Ohgushi Takayuki
Published: January 2005

Abstract.  1. Plant quality can directly and indirectly affect the third trophic level. However, little attention has been paid to how changes in plant quality affect the performance of predators through trophic levels, and which herbivores or predators are affected more strongly by host‐plant quality. The present study examined the effects of artificial cutting of willows on the performance of a willow leaf beetle (Plagiodera versicolora Laicharting) and its predatory ladybird beetle (Aiolocaria hexaspilota Hope).

2. Laboratory experiments showed that performance (survival rate, developmental time, and adult mass) of the willow leaf beetle was higher when fed with leaves of cut willows than when fed with leaves of uncut willows. Performance (developmental time and adult mass) of the predatory ladybird was also improved when it was fed on the leaf beetle larvae that had been fed on leaves of cut willows, compared with those that had been fed on leaves of uncut willows. This indicates that a bottom‐up cascade occurs in the tri‐trophic system.

3. In a comparison of improved performance parameters between the leaf beetle and the ladybird, regenerated willows shortened the developmental time of the willow leaf beetle more than that of the ladybird. This indicates that the impacts of willow cutting on insect performance differ between the second and third trophic levels.
Transcriptional events in a clinical model of oral mucosal tissue injury and repair
Authors: Warburton Gary , Nares Salvador , Angelov Nikola , Brahim Jaime S. , Dionne Raymond A. , Wahl Sharon M.
Published: January 2005

Tissue injury in the oral mucosa activates a cascade of transcriptional events important during the healing process that are not yet clearly defined. To characterize these events and identify potential gene targets for future studies, we used cDNA expression arrays in a clinical model of tissue injury. Mucosal biopsies were taken before third molar extraction, 2–4 hours postoperatively, or at 48 hours. Hybridization patterns were analyzed and validated using real‐time polymerase chain reaction. Prior to extraction, the biopsied mucosal tissues were characterized by a panoply of genes that were constitutively expressed. After injury, analysis revealed differential expression of genes involved in transcription, inflammation, and remodeling. At 2–4 hours after injury, genes such as Fos, Jun, and early growth response protein were up‐regulated, while genes responsible for intercellular adhesion were down‐regulated. At 48 hours after injury, the gene profile had shifted toward tissue remodeling. Here we identify genes constitutively expressed in normal oral mucosa and transcriptional events following mucosal tissue injury, which may be useful in identifying new therapeutic targets.
Original Research Articles – Basic Science: Variable impairment of wound healing in the heterozygous collagenase‐resistant mouse
Authors: Beare Alice H.M. , Krane Stephen M. , Ferguson Mark W.J.
Published: January 2005

Collagen undergoes dramatic reorganization during wound repair. Matrix metalloproteinases degrade and remodel collagen in a tightly controlled process. The collagenase‐resistant mouse, Col1a1tm1Jae, produces type I collagen, which is resistant to degradation by human matrix metalloproteinase 1. These mice grow normally but develop thickened skin with age. We have previously reported that the early wound repair response in homozygous mutant (Col1a1r/r) mice is delayed compared to wild type (Col1a1+/+). However, the late‐stage scar of Col1a1r/r wounds was not significantly altered compared to Col1a1+/+. Here we have investigated the response of heterozygous mice (Col1a1+/r) to wounding, not previously reported. Wound reepithelialization was delayed to a similar degree to wounds in the Col1a1r/r mice. However, the recovery of impaired wound contraction was faster in Col1a1+/r than in Col1a1r/r mice, but still slower than in wild‐type animals. Analysis of wound protein extracts showed expression of some matrix metalloproteinases was prolonged in both the Col1a1r/r and Col1a1+/r wounds compared to wild type. We suggest the partial resistance of collagen to collagenase‐mediated degradation in the heterozygous animals causes equivalent impairment of keratinocyte migration compared to homozygous collagenase‐resistant mice, but that wound contraction during late‐stage healing is only partially retarded.
Transforming growth factor‐β and its effect on reepithelialization of partial‐thickness ear wounds in transgenic mice
Authors: Tredget Eric B. , Demare Jack , Chandran Geethan , Tredget Edward E. , Yang Liju , Ghahary Aziz
Published: January 2005

Transforming growth factor‐β (TGF‐β) is known to affect nearly every aspect of wound repair. Many of the effects have been extensively investigated; however, the primary effect of endogenously derived TGF‐β on wound reepithelialization is still not completely understood. To examine this, two types of wounds were made on a transgenic mouse over‐expressing TGF‐β1. Full‐thickness back wounds were made to compare the wound healing process in the presence of compensatory healing mechanisms. Superficial partial‐thickness ear wounds involving only the epidermis were made to determine the effect of TGF‐β on reepithelialization. In the partial‐thickness ear wounds, at later time points, the transgenic group had smaller epithelial gaps than the wild‐type mice. A greater number of actively proliferating cells, as determined by bromodeoxyuridine incorporation, was also found in the transgenic mice at post‐injury day 8. These results show that TGF‐β1 stimulates the rate of reepithelialization at later time points in partial‐thickness wounds. However, in the full‐thickness back wounds, the transgenic animals exhibited a slower reepithelialization rate at all time points and the number of bromodeoxyuridine‐positive cells was fewer. Our findings would suggest that the overexpression of TGF‐β1 speeds the rate of wound closure in partial‐thickness wounds by promoting keratinocyte migration. In full‐thickness wounds, however, the overexpression of TGF‐β1 slows the rate of wound reepithelialization.
Wound matrix attachment regulates actin content and organization in cells of the granulation tissue
Authors: Carlson Mark A. , Thompson Jon S.
Published: January 2005

Actin cytoskeletal polymerization is associated with a pro‐proliferative, pro‐survival state. We hypothesized that the actin polymerization of wound cells is increased in the presence of wound matrix attachment and is decreased after disruption of this attachment. Musculocutaneous flap and wound splinting models were used to investigate the effect of wound matrix attachment on the actin cytoskeleton. Disruption of wound matrix attachment was accomplished by incision of the wound matrix/dermis interface (wound matrix release) and/or desplinting. Polymerized actin was assayed with phalloidin labeling of wound specimens 24 hours after disruption of attachment and a method to quantify the content and organization of polymerized actin in granulation tissue was used. Disruption of wound matrix attachment decreased the content of polymerized actin, the actin staining intensity, and the actin fiber organization in the granulation tissue of both the flap and splint models. Disruption of wound matrix attachment decreased actin polymerization and fiber organization in the granulation tissue. Our data support the concept that the state of wound matrix attachment regulates the actin cytoskeleton of wound cells.
Migration of keratinocytes is impaired on glycated collagen I
Authors: Morita Keisuke , Urabe Kazunori , Moroi Yoichi , Koga Tetsuya , Nagai Ryuji , Horiuchi Seiko , Furue Masutaka
Published: January 2005

Advanced glycation end products are the chemical modification of proteins induced by sugars in a hyperglycemic condition. Extracellular matrix proteins are prominent targets of nonenzymatic glycation because of their slow turnover rates. The aim of this study was to investigate the influence of nonenzymatic glycation of type I collagen on the migration of keratinocytes. The migration of keratinocytes was dramatically promoted on native type I collagen‐coated dishes compared with that on uncoated dishes. When type I collagen was glycated with glycolaldehyde, large amounts of advanced glycation end products were produced; the glycated collagen I‐coated dishes did not promote the migration of keratinocytes. Glycated collagen I did not affect the proliferative capacity of keratinocytes. However, the adhesion of keratinocytes to glycated collagen I was profoundly diminished in a glycation intensity‐dependent manner. α2β1 integrin is responsible for the migration and adhesion of keratinocytes to type I collagen. Pretreatment with glycated collagen I did not affect the expression level or functional activity of α2β1 integrin on keratinocytes. These findings suggest that in the presence of glycated collagen I, keratinocytes lose their adhesive and migratory abilities. As the glycation did not modify the α2β1 integrin on keratinocytes, it is suggested that glycation may diminish the binding capacity of type I collagen.
LPA‐and PDGF‐Stimulated Contraction of Collagen Matrices Involves Neither by Myosin Light Chain Phosphorylation Nor by MLC Kinase
Authors: Abe M , Ishikawa O , Ho C‐C , Grinnell F
Published: January 2005

Fibroblast‐collagen matrix contraction has been used as a model system to study how cells organize connective tissue. Previous work showed that lysophosphatidic acid (LPA)‐stimulated collagen matrix contraction is independent of Rho kinase whereas platelet‐derived growth factor (PDGF)‐stimulated contraction is Rho kinase dependent. The current studies were carried out to learn more about the molecular motors responsible for LPA‐ and PDGF‐stimulated fibroblast‐collagen matrix contraction. Fibroblasts whose MLC kinase was knocked down using siRNA were used to measure matrix contraction in the presence of LPA or PDGF with or without the Rho kinase inhibitor added. The extent of contraction of MLC kinase‐silenced cells was not detectably different from control cells. Other experiments were carried out to test the effects of LPA and PDGF on MLC phosphorylation with and without Rho kinase inhibitor. After 15 min of growth factor stimulation, levels of diphosphorylated MLC were highest in cells in LPA‐containing medium and lowest in PDGF containing medium. Prior addition of Rho kinase inhibitor markedly reduced phosphorylation in every case. These observations suggested that stimulation of collagen matrix contraction required neither growth factor stimulation of MLC phosphorylation nor MLC kinase.
Recombinant G‐CSF Induces Downregulation of IL‐12 Transcription After Partial Hepatectomy
Authors: Okiyama J , Oishi K , Hayamizu K , Aihaiti X , Ohmori I , Yoshimitsu M , Itamoto T , Asahara T
Published: January 2005

Aims:  Recent studies have demonstrated that administration of recombinant IL‐12 induces cytotoxicity against regenerating hepatocytes. We measured transcription levels of IL‐12 in the remnant liver in a model of partial hepatectomy (PHx) to elucidate the mechanism of already reported facilitation of liver regeneration by recombinant human G‐CSF (rhG‐CSF).

Methods:  After PHx of F344 rats, total RNA of the remnant liver was extracted and mRNA levels of IL‐12p35 and G‐CSF were quantified by RT‐real time PCR.

Results:  G‐CSF mRNA levels increased and IL12p35 mRNA levels decreased at 6 and 20 hours after PHx (P < 0.01). Pre‐PHx rhG‐CSF treatment significantly further down‐regulated IL12p35 mRNA levels at 20 hours (P < 0.01).

Conclusions:  This result suggests that rhG‐CSF pretreatment accelerates tissue repair of the liver at least partially through regulation of cellular immunity.
Dispersion of Pressure to the Head in Brain/NeuroSurgery
Authors: Anzai Chika , Kosidawa Kaoru , Sanpei Rumi , Kouda Masayuki
Published: January 2005

Introduction:  In this field, there are limitations to the positioning of the head in accordance to the postsurgery drain control and respective disorders. Dispersion of pressure to the head is attempted.

Methodology:  Items used: air mat, pressure reducing mat, water‐filled pillow, bath towel folded into quarters, pressure gauge

Experiment subject: 14 patients with no self bodily movements, stable in bed, 14 points or more on the Braden scale at 30 degrees

Procedure: After application of each pressure reducing device on the experiment subject (laying horizontally in the supine position), measure the pressure to the head three times and compute the average value

Results:  Pressure to the head averaged 53.9 mmHg for the towel, mat(s) only. The water‐filled pillow was lower than the others, at 48 mmHg.

Examination:  According to Sanada “in the attempt to lower the pressure to the body, either the weight is lightened, or the area subject to pressure is expanded.” The back of the head, which is an area highly susceptible to pressure sores for the head, the area subject to pressure is confined. It could be said that the conditions for the subjects in this experiment were highly susceptible to pressure sores. Therefore, the frequent transferring of pressure, the use of air mats and water‐filled pillows had been considered as prevention measures; however, with this experiment, the effectiveness of water‐filled pillows was confirmed.

It is viewed that since pressure to the head depends on individual body weight and overall body conditions, at the prevention stage, it is critical to select the pressure reducing device in accordance to the evaluation and assessment.
Basic Fibroblast Growth Factor Accelerates Apoptosis in Acute Incisional Wound Healing and Reduces Scar Formation
Authors: Akasaka Yoshikiyo , Ono Ichiro , Yamashita Toshiharu , Jimbow Kowichi , Ishii Toshiharu
Published: January 2005

Apoptosis has been shown to play an important role in the regulation of wound healing, and growth factors can mediate this process. In this study, we examined the relationship between the degree of healing and the level of apoptosis in full‐thickness‐incisional skin wounds, which were treated by conventional suturing with or without intradermal injection of bFGF (0.1 μg and 1 μg/cm of wound). The width of wound tissue showed that the degree of granulation formation in the 1 μg‐bFGF‐treated group significantly increased on day 7, whereas the degree of scar formation significantly decreased on days 14 and 28. Similarly, apoptotic cells significantly increased in the number on day 4 in the 1 μg‐bFGF‐treated group compared with that of the control group (p = 0.024), and decreased on days 14 and 28. These findings therefore, suggest that the accelerated apoptosis in the bFGF‐treated wounds contributes to the decreased cellularity in inflammatory change through elimination of cells with apoptosis, which resulted also in the reduction of scar formation. It therefore hypothesized that apoptosis is involved in the maturation of an acute wound into scar formation, and that bFGF can accelerate this process.
Interaction Between Human Mesenchymal Stem Cells and Basic Fibroblast Growth Factor or Other Cell Types
Authors: Akino K , Mineta T , Nakagawa H , Akita S
Published: January 2005

The basic fibroblast growth factor (bFGF) is known to proliferate and maintain the adult stem cells. The human mesenchymal stem cells (hMSCs) are very homogenous and constitute of euchromatin nuclei in 10% fetal bovine serum (FBS) and induce the heterochomatin nuclei by bone morphogenetic protein.

In order to further investigate other growth factor, other cell types like bFGF, keratinocytes, dermal fibroblasts and endothelial cells, the cellular and molecular analyses are performed in vitro assay systems.

The various doses of bFGF (0.25–25 μg/ml) significantly proliferate the hMSCs in time‐dependent manner in s serum‐free medium. The ultra‐structure by electron microscopy revealed the very small, round and immature both cytoplasmic and nuclear structure resembling the 10% FBS‐cultured cells. The hMSCs and bFGF and other cells are co‐cultured in 8 μm‐pore chamber systems for 16‐hour incubation. The hMSCs successfully migrate through the pore when keratinocytes, dermal fibroblasts, endothelial cells or bFGF (345.0 ± 68.86, 63.8 ± 16.50, 36.2 ± 9.60, 12.0 ± 4.89, p < 0.01) are placed lower chamber. The clear cellular attachment between hMSCs and keratinocytes is observed in the monolayer co‐culture by electron microscopy.

The hMSCs are stimulated by bFGF and interact with other cell types such as keratinocytes, dermal fibroblasts and endothelial cells in vitro.
Accerelation of Wound Healing with PDWHF and Cultured Cells
Authors: Soejima Kazutaka , Fujisawa Daisuke , Honda Takashi , Isago Tsukasa , Nozaki Motohiro
Published: January 2005

Objective:  Effects of platelet derived wound healing factor (PDWHF)1) with or without cultured cells on angiogenesis in treatment of defected wounds using artificial dermis (AD) were investigated in rat experimental model.

Methods:  Wistar strain rats were used for this study. The PDWHF was prepared from platelets. The endothelial cells and fibroblasts were prepared from thoracic aorta and back skin respectively. Two sites of 2.5 × 2.5 cm full‐thickness wounds were created on back of the animals. Artificial dermis (TERUDERMIS®, TERUMO Co, Japan) were grafted to the wounds. Prior to the AD grafts, following 4 groups were established, control group: AD alone (n = 6), PDWHF group: AD treated with PDWHF (0.1 ml/cm2)(n = 6), cultured cells group: disperse of 1 × 105 cultured endothelial cells and fibroblasts between AD and wound (n = 6) and combination group: combination of PDWHF and cultured cells (n = 6). Five days after surgery, degree of angiogenesis was evaluated by gross inspection and histological study. Evans blue perfusions test was performed to evaluate the degree of new capillary formation in the generated dermis quantitatively.

Results:  The combination group showed vascular invasion into AD 5 days after surgery. In this group, the absorbance of the Evans blue extracted from the grafted dermis was highest among the group.

Conclusion:  The result of the present study revealed that treatment with PDWHF combined with cultured endothelial cells and fibroblasts accelerate the wound angiogenesis. This method may be beneficial to shorten the period between the AD grafts and the secondary skin grafting.
Gene Expression of Liver Regeneration After Partial Hepatectomy in Mice Lacking Type Tumor Necrosis Factor Receptor
Authors: Shimizu Tetsuya , Togo Shinji , Makino Hirochika , Tanaka Kuniya , Nagasima Yoji , Shimada Hiroshi
Published: January 2005

Aim:  To investigate whether TNF‐ is necessary for hepatocyte proliferation, we study liver regeneration after partial hepatectomy in mice lacking TNF receptor‐1.

Methods:  TNF receptor type‐1 knockout mice and wild‐type mice were subjected to two‐thirds partial hepatectomy (PHx). Liver regeneration was evaluated by assessing liver weights and Ki67 immunohistochemistry. Riken cDNA microarray analysis was performed on liver samples from mice undergoing PHx to compare clearly differentiated mouse PHx models (TNFR‐1 knockout mice‐K group, and wild type mice‐W group).

Results:  The cumulative survival after PHx in K group was lower than in W group. The mortality rate in K group during the first 3 days after PHx was higher (33%) than in W group. The time to regain the liver weight in K group was 14 days and 7 days in W group. The plasma IL‐6 levels in K type at 3 hr was significantly higher than in W group. The Ki67 expression in K group at 4 days was lower than in W group. LPS, Toll like receptor 4 precursor and MAPK 8 interacting protein in K group was higher than in W group. For cell cycle‐regulated genes, cyclin D1, NFB light chain and TNF receptor super family membrane 1a in K group was lower than in W group.

Conclusions:  Lack of TNF‐ signaling through TNF receptor type 1 suppresses liver regeneration after partial hepatectomy in spite of enhancement of LPS–JNK pathway, no TNF‐a and IL‐6 pathway.
General session II Moderated by Yashihiro Takami: Down‐Regulation of TFF Expression by TNF‐α in Gastric Epithelial Cells
Authors: Fujii Y , T Shimada , Koitabashi A , Hashimoto T , Hosaka K , Tabei K , Namatame T , Yoneda M , Hiraishi H , Terano A
Published: January 2005

Aim:  Trefoil factor family (TFF) peptides are known to facilitate wound healing in gastric mucosa. However, the regulatory mechanisms of gastric TFF expression are not fully understood yet. In this study, we examined the effect of TNF‐α on TFF1 and TFF2 expression in gastric epithelial cells.

Methods:  MKN45 cells were used. TFF mRNA expression was analyzed by real‐time quantitative RT‐PCR. Promoter sequences of TFF1 gene (−956 to +36) and TFF2 gene (−912 to +24) were inserted into pGL3 vector and reporter gene assays were performed. NF‐κB activity was monitored by using a NF‐κB responsive element‐driven reporter vector.

Results:  (1) TNF‐α(0.1–30 ng/ml) down‐regulated TFF1 and TFF2 mRNA expression in a dose‐dependent manner. (2) Reporter gene assays also confirmed the down‐regulation of TFF1 and TFF2 gene transcription by TNF‐α. (3) TNF‐α activated NF‐κB. (4) Overexpression of dominant negative IκBα prevented both TNF‐α‐induced NF‐κB activation and TNF‐α‐induced down‐regulation of TFF expression.

Conclusions:  TNF‐α down‐regulates gastric TFF expression through NF‐κB pathway, suggesting that TFF expression is sensitive to inflammatory stimuli.
The Expression of Human HtrA1 in Keloid Tissue
Authors: Ikeda Mika , Naito Motoko , Suzuki Shigehiko
Published: January 2005

Aim:  To investigate the expression of human HtrA1 in keloid lesions, and to clarify a possible role of human HtrA1 in keloid pathogenesis.

Methods:  Total RNA was isolated from six keloids and two normal skins by single‐step method. The expression level of human HtrA1 was examined by using Northern blot analysis. Keloid and normal skin tissue samples were fixed in paraformaldehyde, and paraffin sections were obtained. Immunohistochemical analysis was performed with anti‐human HtrA1 polyclonal antibody.

Results:  The mRNA level of human HtrA1 was markedly elevated in keloid samples, compared with normal skin. Using immunohistochemical analysis, fibroblast‐like cells abundantly found in the margin of keloid lesions, were stained with anti‐human HtrA1 antibody. No human HtrA1 staining of fibroblasts in normal skin was observed. Interestingly, no significant staining was detected in hypertrophyic scar lesions, which is a similar dermal disease to keloid.

Conclusion:  Human HtrA1 expression was found to be up‐regulated in keloid lesions, especially in their margin, compared to normal skins and hypertrophic scars. Our data suggest that human HtrA1 could play a critical role in an expression of keloid specific phenotype and in the development of keloid lesions.
Problems of Lower Extremity Amputation Levels
Authors: Tamai Satoru , Inoue Yoshinori , Sugano Norihide , Hirokawa Masayuki , Jibiki Masatoshi , Nakamura Hiroshi , Nakashima Reiko , Iwai Takehisa
Published: January 2005

Objective:  There is no consensus of objective information to determine the indication for lower extremity amputation and the lowest amputation level that will heal. In this report, we studied amputation for the lower extremity ischemia.

Objects and Methods:  We measured transcutaneous oxygen tension (tcPO2) in seven patients who had have ischemic disease of the lower extremity, two patients were in Buerger's disease and five patients were in arteriosclerosis obliterans (ASO). Of these patients, in regard to complications, four patients had diabetes mellitus (DM) and two patients underwent hemodialysis because of chronic renal failure. Six patients of them were men and only one patient was woman, mean age was 59 years old (25∼76 years old).

Results:  Two patients who had Buerger's disease underwent primary amputation. All another five patients who had ASO underwent secondary amputation. Selections of lower extremity amputation levels were above‐knee amputations in three patients, below‐knee amputations in two patients and toe amputation in two patients. There were incurable infections in two patients.

Conclusions:  It is generally said that if patients have had critical limb ischemia without providing hemodynamic improvement, it is necessary for them to be performed lower extremity amputations within 12 months, In concrete terms, segmental Doppler systolic blood pressures are below 50 mmHg and tcPO2 are below 30 mmHg. However in this our study, primary healing rate was 75% when the tcPO2 was above 20 mmHg.
Fetal Dermal Mesenchymal Cells Posses an Ability to Regenerate Dermis in the Reconstitution Model of Scid Mouse
Authors: Kishi Kazuo , Nakajima Hideo , Nakajima Tatsuo
Published: January 2005

Aim:  We have shown that the dermal mesenchymal cells play key roles in the mouse fetal regenerative wound healing. However, the evaluation whether fetal dermal mesenchymal cells posses regenerative capacity in the adult circumstances has not been clarified yet. In the present study, we evaluated the dermal regenerative capacity of fetal dermal mesenchymal cells using reconstitution models on the back of scid mice.

Method:  Fetal dermal and fascial tissues were obtained from C57bl fetal mice at 17 days gestation. Tissues were dissected under microscope. Epidermis and dermis were separated with dispase. The dermis and fascia were digested with trypsin and cells from each tissue were suspended. Either dermal cells or fascial cells were mixed with epidermal cells, and transplanted in the skin defect 7 mm in diameter made on the back of scid mice. The number of cells transplanted was 1 × 107 for each type of cells. Four weeks later, the transplants were histologically examined.

Results:  All of the transplants constituted with dermal cells and epidermal cells regenerate almost normal dermal structures, while transplants mixed with fascial cells and epidermal cells did not regenerate dermis, and left scar like tissues.

Conclusion:  The regenerative capacity of fetal dermal mesenchymal cells was shown even in the adult circumstances.
General session V Moderated by Yashihiko Maehara: Reduction of Ischemia‐Reperfusion Injury to Skin Flaps by Monoclonal Antibody to Leukocyte Endothelial Adhesion Molecule‐1
Authors: Tosa Yasuyoshi , Muramatsu H. , Hukushiyama H. , Otsuka T. , Satoh K. , Hosaka Y.
Published: January 2005

Aim:  Ischemia‐reperfusion (I‐R) injury continues to be a problem for successful free tissue transfer and replantation after prolonged periods of ischemia. It has been shown that leukocytes and vascular endothelial cells release a variety of inflammatory mediators during reperfusion after ischemia. The leukocyte endothelial adhesion molecule‐1 (LECAM‐1) is a ligand on the endothelium for some of the adhesion receptors on leukocytes. The purpose of this study was to evaluate the blockage of leukocyte‐endothelial adhesion by a monoclonal antibody (MAb) to LECAM‐1 in skin flaps to prevent I‐R injury in rats.

Methods:  Male SD rats (225–250 g) were used. A skin flap (45 × 30 mm) supplied by the superficial epigastric A&V including the femoral vessels was isolated unilaterally. The femoral vessels were cross‐clamped the epigastric vessels for 9 hours of ischemia. Animals in treated group received MAb to LECAM −1 i.v. 15 minutes prior to reperfusion; those in the control group received normal saline. Skin flap viability was assessed by tracing the outline of viable and nonviable areas. Data were collected for the following 7 days. These data were corroborated with histological evidence on comparable areas of the flap.

Results:  Tracing analysis revealed average flap survival area of about 90% in treated group and about 20% in control group (p < 0.05). Histopathologically, few inflammatory changes could be observed in treated group, while marked damage was observed in control group.

Conclusions:  From this study, we concluded that treating skin flaps with Mab to LECAM ‐1 was effective for I‐R injury after 9 hours of warm ischemia.
Comparative Study of NonInvasive Vascular Imaging Techniques in the Evaluation of Chronic Venous Leg Ulcers
Authors: Yamaki Takashi , Nozaki Motohiro
Published: January 2005

Aim:  To compare noninvasive vascular imaging techniques in the evaluation of chronic venous leg ulcers, and to detect the discriminating parameters between primary valvular incompetence (PVI) and postthrombotic syndrome (PTS).

Methods:  A total of 61 limbs with chronic venous ulcers were included in this study. Of 61 limbs, 31 had PVI and remaining 30 had documented PTS. The distribution of venous reflux including superficial, deep, and perforating vein was determined by duplex ultrasound. The venous functions including venous volume (VV: ml), venous filling index (VFI: ml/sec), ejection fraction (EF: %), and residual volume fraction (RVF: %) were evaluated by air plethysmography (APG). The near–infrared spectroscopy (NIRS) was used to measure calf muscle total hemoglobin (tHb), oxygenated hemoglobin (O2Hb), and deoxygenated hemoglobin (HHb) levels using APG exercise protocol, and the changes in these parameters were divided into three patterns; Type I: The HHb does not increase beyond the O2Hb during the total procedure. Type II: The HHb increases beyond the O2Hb during the excise. Type III: The HHb increases beyond the O2Hb during the total procedure. Patients with Type III were considered to have most significant calf muscle dysfunction.

Results:  Superficial venous incompetence was significantly predominant in the PVI (p = 0.031). The proportion of deep vein incompetence was significantly higher in the PTS (p = 0.023). Of these, patients with PTS had significantly higher proportion of femoral vein (FV) and popliteal vein (POPV) incompetence compared to these with PVI (p = 0.0006, 0.0015, respectively). There was no statistically significant difference in the proportion of perforator incompetence between the groups (p = 0.611). There were no significant differences in VV (94.8 ± 43.7, 84.7 ± 48.8, p > 0.999), VFI (5.04 ± 2.49, 5.19 ± 3.15, p = 0.236), EF (51.9 ± 19.2, 47.2 ± 25.3, p = 0.903), and RVF (54.8 ± 22.3, 66.1 ± 29.8, p = 0.378). In the NIRS examinations, Type I was found in 20 limbs (64.5%) in the PVI and 3 (10%) in the PTS, and this was a statistically significant (p < 0.0001). On the contrary, Type III was seen in 1 limb (3.2%) in the PVI and 19 (63.3%) in the PTS. The proportion of Type III was significantly higher in the PTS (p < 0.0001).

Conclusions:  These data suggest that reflux in the FV and POPV might play more important role in the development of venous leg ulcers in patients with PTS. The APG–derived parameters did not improve the discrimination power between the two groups. The increased NIRS–derived HHb in the exercising calf muscle is profound in patients with PTS, suggestive of a promising parameter both in the discrimination of the patients with chronic venous leg ulcers and in the follow–up of the patients with deep vein thrombosis.
Presence of Activated Mobile Fibroblasts in Bronchial Alveolar Lavage from Mild Asthmatic Patients
Authors: Larsen K , Nilsson K , Tufvesson E , Malmström J , Wildt M , Andersson A , Malmström A , Löfdahl CG , Marko‐Varga G , Bjermer L , Westergren‐Thorsson G
Published: January 2005

The remodeling process in the lung of asthmatic patients results in a deposition of connective tissue and extracellular matrix components (ECM), which has been described as a subepithelial fibrosis. Fibroblasts, with the phenotypic appearance of myofibroblasts, are considered as the key source of the ECM in the fibrotic tissue. Furthermore, has the ECM molecule biglycan shown to correlate to the hyperactivity of the lung of asthmatic patients. In this study, we report the novel finding of a stretched fibroblast phenotype from bronchoalveolar lavage fluid (BALF) in 30% of the asthmatic subjects (n = 13). No fibroblasts were obtained in BALF from any of the nonasthmatic control subjects (n = 17). These BALF fibroblasts with several characteristics of a myofibroblast displayed increased migratory capacity, accompanied by an induced expression of the small GTPases RhoA and Rac1, when compared to myofibroblasts from corresponding lung biopsies. Data also shows that patients with BALF myofibroblast also have more cells in their lung tissue near to the basement membrane that stain for markers for mesechymal progenitor cells. Moreover, the BALF‐myofibroblasts showed an increase in production of several types of proteoglycans like biglycan. By adding biglycan to normal fibrobalsts it is further possible to mimic the migratory type of myofibroblast found in BALF. To study differences in protein expression pattern between the two phenotypes, we used a gel‐based proteomic technique in combination with MALDI‐TOF mass spectrometry. BALF‐myofibroblast displayed an increase in heat shock protein 20 (HSP20) and myofibroblasts from biopsies displayed an increase in chloride intracellular channel protein (CLIC) and cofilin. These proteins are all known to regulate cell migration by interacting with the actin cytoskeleton. In summery, data demonstrates that biglycan and mesenchymal progenitor cells might have an important role in the early ‐remodeling process in asthmatic patients and are potential future biomarkers for sub‐epithelial fibrosis formation.
Dressing Effects on Cellular Proliferation and Assessment of Fibroblast's Adherence in Vitro
Authors: Bernard FX , Barrault C
Published: January 2005

Introduction:  The aim of this study was to compare the effects of Urgotul® and other greasy dressings and interfaces on normal human dermal fibroblasts (NHDF) monolayers in vitro. The selected end points were the cell proliferation, the morphology of the extracellular matrix (ECM) upon dressing removal, and the structure of the underlying fibroblasts.

Materials and methods:  Equivalent pieces of each dressing were applied on NHDF monolayers for different times. Cell proliferation was measured via [3H] thymidine incorporation. Identical cultures were used to assess the dressing impact on the morphology of cells in culture after MTT staining and micro‐photographing. ECM morphology was shown by immunoflorescence, using an anticollagen I antibody. Effects on cell ultrastructure were documented by confocal laser microscopy after tubulin/actin double labelling.

Results:  Among the five tested greasy dressings and interfaces, only Urgotul® showed a stimulating effect on the proliferation of NHDF. Two dressings did not modify proliferation and two other had cytostatic effects.

In addition, the lesions of the ECM upon dressing removal were clearly the lowest with Urgotul®(low adherence to cellular surface and/or to extracellular matrix). The ultrastructure of the NHDF in direct contact with Urgotul® was not significantly modified.

Conclusion:  Fibroblasts are the key cells of wound healing. The use of nonaggressive greasy gauzes and/or interfaces promoting cell proliferation should be of interest. Clinical evaluations are required to confirm these in vitro results.