bluuebunny/crossref_metadata_2025 huggingface dataset:
Role of nesting resources in organising diverse bee communities in a Mediterranean landscape
Authors: Potts Simon G. , Vulliamy Betsy , Roberts Stuart , O'Toole Chris , Dafni Amots , Ne'eman Gidi , Willmer Pat
Published: January 2005

Abstract.  1. The habitat components determining the structure of bee communities are well known when considering foraging resources; however, there is little data with respect to the role of nesting resources.

2. As a model system this study uses 21 diverse bee communities in a Mediterranean landscape comprising a variety of habitats regenerating after fire. The findings clearly demonstrate that a variety of nesting substrates and nest building materials have key roles in organising the composition of bee communities.

3. The availability of bare ground and potential nesting cavities were the two primary factors influencing the structure of the entire bee community, the composition of guilds, and also the relative abundance of the dominant species. Other nesting resources shown to be important include availability of steep and sloping ground, abundance of plant species providing pithy stems, and the occurrence of pre‐existing burrows.

4. Nesting resource availability and guild structure varied markedly across habitats in different stages of post‐fire regeneration; however, in all cases, nest sites and nesting resources were important determinants of bee community structure.
Density, dispersal, and feeding impact of western flower thrips (Thysanoptera: Thripidae) on flowering chrysanthemum at different spatial scales
Authors: Rhainds Marc , Shipp Les , Woodrow Lorna , Anderson Dale
Published: January 2005

Abstract.  1. This study evaluated the effect of dispersal on the density and feeding impact of a phytophagous insect in relation to the spatial distribution of its host plants.

2. The interaction between density, dispersal, and feeding impact of western flower thrips on flowering chrysanthemum was quantified at three spatial scales, with infested and uninfested plants either isolated in 0.25 m2 individual cages, or enclosed together in 2.25 m2 communal cages or 75 m2 greenhouses.

3. In individual cages, the rate of dispersal from chrysanthemum plants to blue sticky traps increased with the density of thrips for females but not males. Uninfested plants consistently had fewer thrips when they were individually caged rather than enclosed with plants infested with adults, indicating that dispersal mediates inter‐plant distribution of thrips.

4. The feeding impact of thrips on inflorescences was evaluated using the absorbance of ethanol extracts at wavelengths characteristic of yellow carotenoid pigments associated with chrysanthemum inflorescences (415, 445, and 472 nm). Increasing absorbance of extracts with increasing density of thrips per inflorescence suggests that feeding by thrips results in ruptured cells leaching carotenoid pigments.

5. In communal cages, the distribution of thrips was uniform for infested and uninfested plants, whereas the density and feeding impact of thrips in greenhouses were higher for infested than uninfested plants. These results suggest that short‐range dispersal by adults homogenises the density and feeding impact of thrips among host plants only on a small spatial scale.
Flower‐heads, herbivores, and their parasitoids: food web structure along a fertility gradient
Authors: Fonseca Carlos Roberto , Prado Paulo I. , Almeida‐Neto Mário , Kubota Umberto , Lewinsohn Thomas M.
Published: January 2005

Abstract.  1. The ways in which a soil fertility gradient affects three trophic level food webs defined by plants of the family Asteraceae, flower‐head herbivores, and their parasitoids was investigated. It was tested how the fertility gradient alters: (i) the abundance and richness of plants, herbivores, and their parasitoids, (ii) the herbivore–plant ratio, and (iii) the connectance of the plant–herbivore community matrix.

2. From April to May 2000, plants and insects were sampled in 16 Brazilian Cerrado (sensu stricto) sites along a physiognomic gradient varying from open shrublands (cerrado) to closed woodlands (cerradão). Sites were objectively positioned along the physiognomic gradient by a single index, tree density. Sixty‐seven per cent of the variation in tree density among sites was correlated to two principal components of a PCA, representing gradients of soil fertility.

3. Asteraceae abundance, richness, and flower‐head availability were negatively related to tree density due to their preference for sunny environments, despite the surplus of soil nutrients. The abundance and richness of Diptera and Lepidoptera, the most important flower‐head herbivores, were also negatively related to tree density. Parasitoid abundance decreased with tree density; however, the number of parasitoids per hosts was lower in cerrado, suggesting that top‐down forces are not getting stronger in more productive sites, as could be expected.

4. Community allometry analyses showed that the herbivore to plant ratio was independent of community richness and did not respond to tree density.

5. Connectance of the plant–herbivore matrix was dependent on the community matrix size. Proportionally, species‐rich cerrado sites had fewer interactions than their species‐poor counterparts. Nevertheless, after removing the effect of the matrix size, connectance was not related to tree density.

6. Soil fertility, as the primary cause of the cerrado–cerradão physiognomic gradient, strongly affected the abundance and richness of plants, herbivores and their parasitoids; however, it had little effect on important community attributes, such as the herbivore–plant ratio and the connectance of the plant–herbivore matrix.
Pollution by conspecifics as a component of intraspecific competition among Aedes aegypti larvae
Authors: Bédhomme Stéphanie , Agnew Philip , Sidobre Christine , Michalakis Yannis
Published: January 2005

Abstract.  1. The role of pollution by conspecifics in the costs associated with larval intraspecific competition was investigated for Aedes aegypti (Diptera: Culicidae).

2. The growth of larval A. aegypti mosquitoes reared in clean water and water in which another larva had previously grown was compared; this procedure eliminates interactions through food consumption between competitors and allows the effects of other processes to be expressed.

3. A cost of growing in polluted water was found: this cost was expressed as an increase in developmental time and a reduction of adult longevity when starved, starved adult dry weight, and wing length.

4. Contrary to previously reported results of an experiment allowing for competition for food, these costs were not expressed in a sex‐specific manner and were independent of the sex of the polluter.

5. It was thus demonstrated that competition arises from both resource depletion and other effects that result in deterioration of the environment, with chemical pollution of the environment being the most likely cause.
Direct and indirect interactions involving ants, insect herbivores, parasitoids, and the host plantBaccharis dracunculifolia(Asteraceae)
Authors: Fagundes Marcílio , Neves Frederico S. , Fernandes G. Wilson
Published: January 2005

Abstract. 1. The relative importance of direct and indirect interactions in controlling organism abundance is still an unresolved question. This study investigated the role of the direct and indirect interactions involving ants, aphids, parasitoids, and the host plantBaccharis dracunculifolia(Asteraceae) on a galling herbivoreBaccharopelma dracunculifoliae(Homoptera: Psyllidae).

2. The effects of these interactions on the galling herbivore's performance were evaluated by an exclusion experiment during two consecutive generations of the galling insect.

3. Ants had a direct negative effect on the performance of the galling herbivore by reducing the number of nymphs per gall. In contrast, ants had no indirect effects on gall mortality through the associated parasitoids.

4. Aphids negatively affected gall development, suggesting that galls and aphids might be partitioning photoassimilates and nutrients moving throughout host‐plant tissues.

5. In addition, galls that developed during the rainy season were heavier, indicating that variation in the host plant, due to weather changes, can affect the development ofB. dracunculifoliaegalls. However, variation in the development ofB. dracunculifoliaegalls due to presence of aphids or the weather changes did not affect parasitoid attack.

6. These results suggest that direct interactions between ants and galls influenced galling insect abundance, whereas numerical indirect effects involving galling insects, ants, aphids, and host plants were less conspicuous.
Interactions between granivorous and omnivorous ants in a desert grassland: results from a long‐term experiment
Authors: Valone Thomas J. , Kaspari Michael
Published: January 2005

Abstract.  1. There is growing evidence that granivorous ants compete for seeds in desert ecosystems. But ants that diverge in diet may still interact strongly if existing colonies deter foundress queens. Granivores (seed eaters) and omnivores (ants that scavenge and feed on plant exudates) are common components of desert ecosystems.

2. The results of a 15‐year granivore reduction experiment to explore the responses of two common omnivorous ants, Forelius cf pruinosus and Dorymyrmex insana, in a desert grassland were analysed. The food competition hypothesis predicts that granivore removal will have no net effect on omnivores. The spatial competition hypothesis predicts compensatory increases in omnivores on granivore removal plots.

3. A grain‐based gut poison significantly reduced the densities of three of the four common granivorous ants. Densities of the large‐colony (105‐worker) omnivore Forelius did not increase with granivore removal. In contrast, and consistent with the hypothesis of competition for space, densities of the small‐colony omnivore (102−103 worker) Dorymyrmex increased on granivore removal plots; however, this effect was only observed in years of high Dorymyrmex abundance.
Does differential predation permit invasive and native mosquito larvae to coexist in Florida?
Authors: Griswold Marcus W. , Lounibos L. Philip
Published: January 2005

Abstract. 1. The hypothesis that selective predation on larvae of the invasiveAedes albopictus(Skuse) could account for its stable coexistence with the native mosquito species and inferior competitorOchlerotatus triseriatus(Say) in Florida treeholes and container systems was tested experimentally.

2. Functional responses of the two dipteran predatorsToxorhynchites rutilus(Coquillett) andCorethrella appendiculata(Grabham) were evaluated separately forA. albopictusandO. triseriatusprey. Both predators exhibited type II functional responses and consistently consumed more of the invasive species. Handling time ofT. rutilusfeeding uponO. triseriatuswas significantly longer than when preying upon the invasive species.

3. When either predator species was offered varying ratios of the two prey species,A. albopictuswas consumed preferentially. The absence of a prey ratio effect on preference indicated that switching probably does not occur.

4. The higher maximum feeding rate upon, and preference for,A. albopictussuggests that differential predation may foster coexistence of the invasive and native mosquito prey species in Florida.
Age‐dependent clutch size in a koinobiont parasitoid
Authors: Elzinga Jelmer A. , Harvey Jeffrey A. , Biere Arjen
Published: January 2005

Abstract.  1. The Lack clutch size theory predicts how many eggs a female should lay to maximise her fitness gain per clutch. However, for parasitoids that lay multiple clutches it can overestimate optimal clutch size because it does not take into account the future reproductive success of the parasitoid.

2. From egg‐limitation and time‐limitation models, it is theoretically expected that (i) clutch size decreases with age if host encounter rate is constant, and (ii) clutch size should increase with host deprivation and hence with age in host‐deprived individuals.

3. Clutch sizes produced by ageing females of the koinobiont gregarious parasitoid Microplitis tristis Nees (Hymenoptera: Braconidae) that were provided daily with hosts, and of females ageing with different periods of host deprivation were measured.

4. Contrary to expectations, during the first 2 weeks, clutch size did not change with the age of the female parasitoid, neither with nor without increasing host‐deprivation time.

5. After the age of 2 weeks, clutch size decreased for parasitoids that parasitised hosts daily. The decrease was accompanied by a strong decrease in available eggs. However, a similar decrease occurred in host‐deprived parasitoids that did not experience egg depletion, suggesting that egg limitation was not the only factor causing the decrease in clutch size.

6. For koinobiont parasitoids like M. tristis that have low natural host encounter rates and short oviposition times, the costs of reproduction due to egg limitation, time limitation, or other factors are relatively small, if the natural lifespan is relatively short.

7. Koinobiont parasitoid species that in natural situations experience little variation in host density and host quality might not have strongly evolved the ability to adjust clutch size.
Perspective Article: Tissue repair, contraction, and the myofibroblast
Authors: Desmoulière Alexis , Chaponnier Christine , Gabbiani Giulio
Published: January 2005

After the first description of the myofibroblast in granulation tissue of an open wound by means of electron microscopy, as an intermediate cell between the fibroblast and the smooth muscle cell, the myofibroblast has been identified both in normal tissues, particularly in locations where there is a necessity of mechanical force development, and in pathological tissues, in relation with hypertrophic scarring, fibromatoses and fibrocontractive diseases as well as in the stroma reaction to epithelial tumors. It is now accepted that fibroblast/myofibroblast transition begins with the appearance of the protomyofibroblast, whose stress fibers contain only β‐ and γ‐cytoplasmic actins and evolves, but not necessarily always, into the appearance of the differentiated myofibroblast, the most common variant of this cell, with stress fibers containing α‐smooth muscle actin. Myofibroblast differentiation is a complex process, regulated by at least a cytokine (the transforming growth factor‐β1), an extracellular matrix component (the ED‐A splice variant of cellular fibronectin), as well as the presence of mechanical tension. The myofibroblast is a key cell for the connective tissue remodeling that takes place during wound healing and fibrosis development. On this basis, the myofibroblast may represent a new important target for improving the evolution of such diseases as hypertrophic scars, and liver, kidney or pulmonary fibrosis.
Original Research Articles – Clinical Science: Annual costs of treatment for venous leg ulcers in Sweden and the United Kingdom
Authors: Ragnarson Tennvall Gunnel , Hjelmgren Jonas
Published: January 2005

The aim of this study was to estimate costs of treating venous leg ulcers in Sweden and the United Kingdom during 1 year and to quantify costs in different health states. The costs of treating four different types of venous leg ulcers were estimated for 52 weeks by a stochastic health economic model, which simulated resource use data obtained from prospectively collected patient data, expert panels in the two countries, and published scientific literature. The average cost of treating an ulcer varied between €1332 and €2585 in Sweden and from €814 to €1994 in the United Kingdom. Cost of treating large ulcers (≥ 10 cm2) of long duration (≥ 6 months) was highest in both countries. Frequency of dressing changes and duration of time for each dressing change were higher in Sweden than in the United Kingdom, resulting in higher total cost per patient in Sweden. An important factor for the total costs was time to heal. Other important variables influencing treatment costs were frequency and duration of dressing changes. Actions to reduce time used for dressing changes and the total time to healing are thus very important in reducing costs spent on treatment of venous leg ulcers in both countries.
Expression of different p63 variants in healing skin wounds suggests a role of p63 in reepithelialization and muscle repair
Authors: Bamberger Casimir , Hafner Annina , Schmale Hartwig , Werner Sabine
Published: January 2005

Healing of skin wounds in mammals involves partial reconstruction of the dermis and coverage of the injured site by keratinocytes. The latter process is achieved by extensive migration and hyperproliferation of keratinocytes at the wound rim. Because the p53 protein family member p63 is expressed in human hyperproliferative epidermis, this study determined whether enhanced keratinocyte proliferation correlates with the expression of p63. Therefore, we investigated the temporal and spatial distribution of four major variants of the p63 transcription factor—TAp63α, TAp63γ, ΔNp63α and ΔNp63γ—during normal skin wound healing in mice. Transcripts encoding amino‐terminally truncated ΔNp63 variants were found at high levels in basal and suprabasal keratinocytes of the hyperproliferative wound epithelium. Interestingly, TAp63 variants, which include the conserved transactivation domain TA at their amino‐terminus, were also expressed in wound keratinocytes as well as at the edge of the injured subcutaneous muscle panniculus carnosus. These findings suggest splice‐variant specific functions of p63 in reepithelialization and muscle repair.
In vivo gene delivery of Ad‐VEGF121 to full‐thickness wounds in aged pigs results in high levels of VEGF expression but not in accelerated healing
Authors: Vranckx Jan J. , Yao Feng , Petrie Nicola , Augustinova Hanka , Hoeller Daniela , Visovatti Scott , Slama Jarek , Eriksson Elof
Published: January 2005

We have previously reported that endogenous vascular endothelial growth factor (VEGF) concentration in older pig wounds peaked later and at one‐fourth the level of young pigs. These data suggested that VEGF might play a major role in the healing of full‐thickness wounds in the aged pig. By in vivo gene transfer using the microseeding technique, we treated full‐thickness wounds with different doses of VEGF‐expressing adenoviral vector (Ad‐VEGF) varying from 1 × 107 to 2.7 × 1011 particles per wound (ppw). We found that the VEGF expression in wound fluid followed a dose–response pattern. However, when wounds were microseeded with the highest concentration of Ad‐VEGF (2.7 × 1011 ppw), diminished healing rates were found. We then determined the minimal functional concentrations of Ad‐VEGF. We used five aged Yucatan minipigs, all retired breeders, to analyze the role of over‐expression of 1 × 108 and 1 × 109 ppw of Ad‐VEGF (n= 78) in terms of healing of full‐thickness wounds, all 2.5 × 2.5 × 1 cm in size (n= 158). The Ad‐VEGF solutions were delivered to the wound floor and borders by in vivo microseeding. Control wounds (n= 80) were microseeded with Ad‐Lac‐Z (n= 25), treated with saline (n= 49) or treated dry (n= 6). All wounds except for the dry‐treated ones were covered with a wound chamber and a wet environment was created by injecting 2.5 ml saline into the chamber. Peak VEGF expression (2300–4000 pg/ml) was detected on days 2 or 3 post gene delivery. This level of VEGF expression was not seen in the saline (n= 49) or Ad‐null (n= 25) control groups. The VEGF expression in wounds treated with 1 × 108 and 3 × 108 ppw (n= 39) exhibited a slower onset with a peak concentration of 400–920 pg/ml on days 5–7. Although high levels of VEGF expression were achieved in the local wound environment, we could not show a significant increase in neovascularization as compared to saline‐treated wounds. No significant differences were observed in the rate of reepithelialization and wound contraction among groups of full‐thickness wounds treated with Ad‐VEGF, Ad‐null mutant, or saline in the aged “wet wound healing” pig model. These results indicate that increased levels of VEGF in wounds produced by in vivo gene transfer have little effect on the healing of full‐thickness wounds in the aged pig model. Moreover, significantly higher levels of VEGF expression by Ad‐VEGF could lead to impaired wound healing.
Ontogeny of expression of transforming growth factor‐β and its receptors and their possible relationship with scarless healing in human fetal skin
Authors: Chen Wei , Fu Xiaobing , Ge Shili , Sun Tongzhu , Zhou Gang , Jiang Duyin , Sheng Zhiyong
Published: January 2005

Fetal cutaneous wounds that occur in early gestation heal without scar formation. Although much work has been done to characterize the role of transforming growth factor‐β (TGF‐β) isoforms and their receptors in the wound healing process, their roles in scarless wound repair observed in early gestation and their functions in human fetal skin development, and structural and functional maintenance are still not well understood. In this study, we explore the expression and distribution characteristics of three TGF‐β isoforms and their receptors, TGF‐βRI (TBRI) and TGF‐βRII (TBRII), in fetal and postnatal skins to understand the relevance of these five proteins to skin development and elucidate the mechanism(s) underlying the phenotypic transition from scarless to scar‐forming healing observed during fetal gestation. Fetal skin biopsies of human embryo were obtained from spontaneous abortions at different gestational ages from 13 to 32 weeks and postnatal skin specimens were collected from patients undergoing plastic surgery. Gene expression and positive immunohistochemical signals of TGF‐β1, TGF‐β2, TGF‐β3, TBRI, and TBRII could all be detected in fetal and postnatal skins. In early gestation, gene expression of TGF‐β1, TBRI, and TBRII was weaker and protein contents were less compared with postnatal skins (p < 0.05). In contrast, more TGF‐β2 mRNA transcript was found in early gestation than in late gestation and in postnatal skins, whereas protein content of this growth factor increased during gestation. Lastly, mRNA transcript and protein contents of TGF‐β3 were apparently higher in early gestation compared to postnatal skin (p < 0.05). In postnatal skin, granules containing the three TGF‐β isoforms were mainly distributed in the cytoplasm and extracellular matrix of epidermal cells, interfollicular keratinocytes, and some fibroblasts. TBRI and TBRII were chiefly located in the cellular membrane of epidermal keratinocytes and some fibroblasts. The endogenous three TGF‐β isoforms and their receptors may be involved in the development of embryonic skin and in the maintenance of cutaneous structure and function, and also in postnatal wound healing. The differential levels of TGF‐β isoforms may provide either a predominantly antiscarring or profibrotic signal upon wounding depending on the gestational period. Lower expression of their receptors in early gestational skins may be a reason for the reduced ability to perceive ligands, ultimately leading to scar‐free healing.
Cultured pressure ulcer fibroblasts show replicative senescence with elevated production of plasmin, plasminogen activator inhibitor‐1, and transforming growth factor‐β1
Authors: Vande Berg Jerry S. , Rose Michael A. , Haywood‐Reid Patricia L. , Rudolph Ross , Payne Wyatt G. , Robson Martin C.
Published: January 2005

In a 16‐patient study, cultured fibroblast populations from normal skin were able to replicate an average of 14.8 ± 2.2 times before becoming senescent, while fibroblast populations from the ulcer bed reached the end of their replicative life span after 7.2 ± 1.9 population doublings (p= 0.001). Fibroblast populations from 10 of 16 pressure ulcers became senescent after fewer than five population doublings, whereas when populations of fibroblasts from adjacent normal skin were studied, only 2 of 16 became senescent within this same time period. In addition, only an occasional fibroblast from normal skin stained positively for senescence‐associated β‐galactosidase compared to approximately 50% of equally aged ulcer bed fibroblasts (p = 0.0060). Senescent ulcer bed fibroblasts secreted significantly more plasmin than early passage ulcer bed fibroblasts (p= 0.0237), nearly six times as much plasmin as early passage normal skin fibroblasts (p < 0.0001), three and a half times the level of normal skin fibroblasts of the same age (11.52 ± 4.58 µg/mg protein; p= 0.0003), and more than one and a half times the level of senescent normal skin fibroblasts (p= 0.0525). Senescent pressure ulcer fibroblasts generated significantly more plasminogen activator inhibitor‐1 (1179.27 ± 25.37 ng/mg protein) than normal skin fibroblasts of the same age (132.16 ± 16.20 ng/mg protein; p = 0.0357). Also, senescent ulcer bed fibroblasts produced higher levels of transforming growth factor‐β1, but these were not significantly different from senescent normal skin fibroblasts. Although senescent ulcer fibroblasts produce elevated levels of plasminogen activator inhibitor‐1 and transforming growth factor‐β1, the ratio of these factors to plasmin levels suggests that this may have little influence on extracellular matrix synthesis or maintenance in the chronic wound. These data show that cultured fibroblasts from most patient pressure ulcers profile a wound environment that is associated with an increasing population of senescent fibroblasts; however, factors within the chronic wound environment that promote cellular senescence remain unclear. We have proposed that a prolonged inflammatory response may be a contributing factor to the chronic wound condition.
Original Research Articles – Regeneration Science: Epidermal stem cells are the source of sweat glands in human fetal skin: Evidence of synergetic development of stem cells, sweat glands, growth factors, and matrix metalloproteinases
Authors: Fu Xiaobing , Li Jianfu , Sun Xiaoqing , Sun Tongzhou , Sheng Zhiyong
Published: January 2005

The development of sweat glands is a complex biological process, and the extent of cellular trafficking between epidermal stem cells and the development of sweat glands is uncertain. Therefore, we studied the synergetic development effects of stem cells, sweat glands, growth factors, and matrix metalloproteinases (MMPs) in human skin. Human fetal skin was obtained from spontaneously aborted fetuses at 11–31 weeks of gestation. Paraffin sections were cut and stained with hematoxylin and eosin or immunostained with antibodies against β1 integrin, keratin (K)‐19 and K7, MMP‐2 and ‐7, and epidermal growth factor. In situ hybridization was used along with semiquantitative analysis of the positive expression of these proteins to analyze for mRNA expression of MMP‐2 and ‐7. Histological studies revealed the fetal epidermis began to form a primary epidermal ridge at gestational age 13–14 weeks and these primordial basal cells became tightly packed to take the form of multiple hillocks between 14 and 16 weeks. Furthermore, these cells gave rise to chord‐like columnar buds in the embryonic epidermis, and these buds gradually migrated downward into the dermis to form juvenile sweat glands at 18–20 weeks. Mature sweat glands were found in the fetal epidermis at the end of 24 weeks. β1 integrin and K19 immunoreactivities were first detected in those cells that gathered together to form primary epidermal ridges, including sweat gland cords, buds, and immature sweat gland cells. The positive immunostaining for K7 appeared in early sweat gland buds at 14–16 weeks, and from then on K7 was concentrated in developing sweat gland cords or cells. At 14–16 weeks, positive epidermal growth factor, MMP‐2, and MMP‐7 expression was first observed weakly in developing sweat gland buds. The immunoreactivity of these proteins was then gradually increased in the developing sweat gland buds and extracellular stroma from 14 to 20 weeks. The intensity of the positive signal peaked at 20–22 weeks of gestational age. After that, the intensity of immunostaining for MMP‐2 and MMP‐7 proteins was gradually weakened. However, the expression of epidermal growth factor did not show an apparent decrease. These results suggest that epidermal stem cells are the source of sweat glands. Epidermal growth factor is one of the main inducers in the development and maturity of sweat gland buds or cells and the local activated MMPs may play an important role in cleaving the major matrix components in the basement membrane.
Involvement of bone marrow‐derived cells in healing of experimental colitis in rats
Authors: Komori Masato , Tsuji Shingo , Tsujii Masahiko , Murata Hiroaki , Iijima Hideki , Yasumaru Masakazu , Nishida Tsutomu , Irie Takanobu , Kawano Sunao , Hori Masatsugu
Published: January 2005

Bone marrow is reported to contain hematopoietic stem cells and other adult somatic stem cells that have phenotypes of cells composing tissues other than bone marrow. To explore the implication of bone marrow‐derived cells in the treatment of inflammatory bowel diseases, experimental colitis was induced in wild‐type rats after transplantation of bone marrow from transgenic rats expressing green fluorescence protein (GFP). Chronic colitis was induced 21 days later using 30 mg 2,4,6‐trinitrobenzenesulfonic acid (TNBS). Control rats received saline. At 28, 56, and 224 days after TNBS administration, rats were euthanized, and tissues were removed and processed for paraffin‐embedded sections. Cells derived from bone marrow were identified by immunohistochemistry using anti‐GFP antibody. To identify the phenotypes of the cells expressing GFP, we conducted serial‐section analysis and double‐staining analysis using antibodies against cytokeratin (epithelial cells) or vimentin (interstitial cells). In the present study, GFP‐positive, bone marrow‐derived cells occupied 37.6% and 4.25% of the colonic epithelium at 28 days and 56 days after the induction of TNBS‐colitis, respectively. Also, significant amounts of mucosal and submucosal interstitial cells were derived from the bone marrow. These findings showed that a large amount of bone marrow‐derived cells were involved in regeneration of the colon after experimental colitis in rats.
General session I Moderated by Yashihiro Otani: A Case of Lower Leg Osteomyelitis Successfully Treated with Trafermin Spray
Authors: ENDO Takashi
Published: January 2005

Aim:  Chronic osteomyelitis with pus discharge is very difficult to treat. This time we experienced the case of lower leg osteomyelitis, which we have treated with Trafermin spray. The result was excellent and beyond our expectation, so we will report this case.

Case:  63 years, male. He sustained the open fracture injury of his right foot by a fallen rocks and resulted in the osteomyelitis. Then he was introduced to our hospital. Operation was done under spinal anesthesia, the infected bone and covering skin were excised fully and the defect (4 × 3 × 3 cm) was left open. After 2 weeks of dressing change, we began to use the Trafermin spray.

Result:  At the first 3 weeks, evident change was not recognized, but good granulation formation started 4 weeks later. This tendency has continued for a while and the defect completely closed 5 months later.

Discussion:  We have experienced the case of lower leg osteomyelitis, which showed the dramatic improvement. Through this case, we have reconfirmed that rich blood supply around the defect is essential for the healing process when we use the Trafermin spray.
NonInvasive Approach for Refractory Enterovaginal Fistula
Authors: Hidefumi Nishimori , Hata Fumitake , Furuhata Tomohisa , Ishiyama Gentaro , Hirata Koichi
Published: January 2005

Enterovaginal fistula is generally difficult to treat, and a surgical treatment is often chosen. There are many cases that require extended bowel resection for severe adhesion in the pelvic cavity and just as many cases of postoperative complications. The present case involves a 61‐year‐old female. After radical operation for ovarian cancer, a recurrence was recognized in the pelvic cavity despite chemotherapy. After that, recurrence was found in the pelvic cavity, and low anterior resection combined with the resection of recurrent tumor was performed.

Enterovaginal fistula and pelvovaginal fistula were found from the 10th postoperative day; therefore urethral catheter was placed transvaginally. Those fistulas were simplified afterwards, and the tube for percutaneous transhepatic cholangio drainage (PTCD) was placed and then removed after several exchanges. After that, enteric juice was identified for a few days, but the fistula was completely closed, and no recurrence was found.

It is easy for discharge from enterovaginal fistula to increase because of the anatomical position the fistula; since there is no appropriate device for this fistula, treatment is difficult. It seems that, in selected cases in which the patient heals conservatively without losing quality of life for patients, it is possible to place a tube in an appropriate position under the x‐ray and change it step by step.
Attempt to Improve Wound Healing Considering the Timing of Suture Removal and the Dressing
Authors: Kitte Toshihiro , Arinaga Shinya , Ishikawa Koichi , Shiraishi Norio , Kitano Seigo
Published: January 2005

Introduction:  In the surgical field, “suture removal on 7 days after surgery” and “daily disinfection and coverage with gauze” have been customs. The timing of suture removal and the postoperative wound treatment were devised, and their effect on wound healing was compared with the conventional method.

Method:  The abdominal incision was divided into two parts, one half of the suture was removed on 3 days after surgery, the other half of the suture was removed on 7 days after surgery, and the conditions of the wound were compared between 7 and 14 days after surgery. In addition, the skin was disinfected only immediately before incision, and the closed dressing was given on the wound.

Results:  There were no differences in the degree of skin approximation, the degree of redness, subcutaneous swelling, pain and postoperative scar between both days. However, the suture removal on 3 days gave no scar of suture, which was better cosmetically. The frequency of treatments also decreased, which led to cost reduction.

Discussion:  Since the wound is covered with epithelial cells in 48 hours, the suture removal on 3 days and the closed dressing are desirable in the abdominal incision to which no especially strong stress is added from the viewpoint of wound healing. In addition, this method is better than the conventional method from the aspect of cosmetics and cost reduction. We will investigate this method further in the future.
symposium i‐2. pathophysiology and treatment of pressure ulcer Moderated by Toshimasa Asahara: Sterilized Acellular Allogeneic Dermal Matrix for Scaffold of Cultured Composite Skin
Authors: Guo Jingping , Takami Yoshihiro , Yamaguchi Ryo , Shiota Noriko , Ogo Ken , Harii Kiyonori
Published: January 2005

Aim:  Acellular Allogeneic Dermal Matrix ADM has been used for scaffold of cultured composite skin. However, possibility of disease transmission by ADM remains as its potential risk. So it is important to reduce the risk by ADM transplantation the purpose of this study was to evaluate whether ADM could be used as a scaffold of cultured composite skin safely.

Method:  Four different kinds of methods were used for sterilization, such as 98% glycerol, Pasteurization, Gamma irradiation, and ethylene oxide gas sterilization (EOG). Histological, immunohistochemical and electrophoretic (SDS‐PAGE) properties were evaluated. The property as a scaffold of cultured composites skin was evaluated using normal human cultured keratinosytes.

Results:  Four methods did not induce apparent collagen degradation of ADM on SDS‐PAGE. However, damage of basement membrane structure was observed by ADM treated with Pasteurization, Gamma irradiation, and EOG histological. These three methods also resulted in the poor regeneration of the epidermal layer in cultured composite skin.

Conclusion:  The results suggested that glycerol treatment was found to be the best method of ADM sterilization as far as its scaffold was concerned.
Regulation of VEGF‐Induced Angiogenesis by MMPs
Authors: Shiomi Takayuki , Hashimoto Gakuji , Inoki Isao , Fujii Yutaka , Ikeda Eiji , Okada Yasunori
Published: January 2005

Aim:  We have recently reported that connective tissue growth factor (CTGF) inhibits the angiogenic activity of VEGF165, one of the vasucular endothelial growth factor (VEGF) isoforms, through the complex formation with VEGF165. In the present study, we examined the susceptibility of the VEGF165/CTGF complex to matrix metalloproteinases (MMPs), ADAMTS4 and serine proteinases, and evaluated the recovery of the angiogenic activity of VEGF165 after the treatment.

Methods:  VEGF165/CTGF complex was digested with six different MMPs, ADAMTS4, elastase or plasmin. The reaction products were analyzed on silver‐stained gels and by immunoblotting. Angiogenic activity of the complex treated with MMPs was assayed by in vitro tube formation assay and in vivo angiogenesis assay using a Matrigel injection model in mice.

Results:  Among the MMPs, MMP‐1, ‐3, ‐7, and ‐13 processed CTGF of the complex into the major NH2‐ and COOH‐terminal fragments, whereas VEGF165 was completely resistant to the MMPs. The in vitro angiogenic activity of VEGF165 blocked in the VEGF165/CTGF complex was reactivated to original levels after CTGF digestion of the complex with MMPs. Recovery of activity was further confirmed by in vivo angiogenesis assay.

Conclusions:  These results demonstrate that CTGF is a substrate of MMPs and that the angiogenic activity of VEGF165 suppressed by the complex formation with CTGF is recovered through the selective degradation of CTGF by MMPs. MMPs may play a novel role through CTGF degradation in VEGF‐induced angiogenesis during formation of granulation and scar tissue in wound healing process.
Skin Regeneration by Human Mesenchymal Stem Cells and Basic Fibroblast Growth Factor
Authors: Nakagawa H , Akino K , Akita S
Published: January 2005

Skin and soft tissue defects are sometimes problematic especially when the defects large, contaminated, irradiated, or poor blood supplied. The human mesenchymal stem cells (hMSCs) are proliferated upon basic fibroblast growth factor (bFGF) stimuli in vitro and in vivo. In this experiment, the skin and soft tissue defects are investigated if the wounds are able to be reepithelialized or accelerated by hMSCs, bFGF and porcine‐derived bilayered skin template.

1.5 × 1.5 cm2 nude rat skin and soft tissue defects including panniculus carnosus are excised and 1 × 106 hMSCs and various doses of bFGF (1–100 μg) applied. Before and after normal reepithelialization, the tissues are tested for protein expressions by immunohistochemistry and Western blotting.

The wound sizes are significantly decreased at day 7 with hMSCs with 1, 10, or 100 μg bFGF compared to hMSCs‐alone or medium‐only. All the wounds healed by day 42. 42 Kda and 38 Kda human‐derived pancytokeratin expressions, which do not cross‐react with murine antigens, by Western blotting significantly augmented by 10 μg bFGF compared to hMSCs‐alone. The epidermal immunolocalizations such as integrin α3 and SKALP (Skin‐derived Anti Leukoproteinase) are greatly elevated in time and dose‐dependent manner. Human pan‐cytokeratin expressions are immunoreactive even at day 42.

These data suggest the skin and soft tissue wound healing is accelerated by hMSCs together with bFGF, partly by means of differentiation of hMSCs toward epidermal components.
Symposium i‐2. pathophysiology and treatment of pressure ulcer Moderated by Hiroyuki Kuwano: Administration of Basic Fibroblast Growth Factor (bFGF) for Intractable Rectovesical Fistula — a Case Report
Authors: Onozato Ryoichi , Yamaguchi Satoru , Asao Takayuki , Kobayashi Tsutomu , Tsutsumi Soichi , Kuwano Hiroyuki
Published: January 2005

Case:  62‐year‐old female. Drainage of abscess and hysterectomy were performed for pelvic peritonitis. But stool‐like exudates was observed at postoperative 2 days. Transverse colostomy and drainage of the abscess were performed. Transvaginal closure of the vaginal stump was performed for rectovaginal fistula. After a while, intractable rectovesical fistula was newly observed.

Clinical course:  Because patient was contracted articular rheumatism, she wanted to avoid operation. Under sufficient informed consent, we started the treatment of injection with bFGF via the drainage tube to rectovesical fistula (50 μg/day). Narrowness of the fistula was checked by fistulography, and decrease of eduction of stool like exudates was observed. The fistula was closed after 2.5 months from beginning of the treatment.

Summary:  We reported a case that bFGF was available for intractable nontumorous enterogenic fistula.
Open Wound Integrated Regenerative Therapy with Topical bFGF Trafermin for Deep Soft Tissue Defects with Bone Exposure
Authors: Ohura N , Ichioka S , Asano T , Nakatsuka T
Published: January 2005

Aim:  Recently advocated concept of wound bed preparation (WBP) has systematically concreted the critical components of wound care, including debridement, bacterial balance, and management of exudate. We succeed in experiences by topical pharmaceutical therapy combined with WBP. We aimed to demonstrate availability of the wound management.

Methods:  Since 1999, 18 exteriorized‐bone patients with deep soft tissue defects were treated with this treatment. Our procedure was composed of four steps; 1) Resection or debridement, 2) Wound cleansing (soaking foot and hand in the hot bath with antibacterial carbonated agent), 3) Topical medication: basic fibroblast growth factor (Trafermin; KAKEN, Tokyo) and bucladecine sodium (Actocin ointment; Daiichi Pharmaceu. Co. Ltd, Tokyo), and 4) Dressing.

Results:  The average time for whole coverage of the exposed‐bone area with healthy granulation was 23 ±  13( median;19, range; 7–47) days after surgical treatment (the 1st step). The average time for complete epithelization was 72 ± 42 (55, 26–150) days in the patients who had not undergone skin graft. For minor amputations of diabetic feet the healing time until complete epithelization was 65 ± 46 (44, 26–150) days on an average.

Conclusions:  We concluded that the integrated approach mobilizing commercial available products for wound healing under optimally prepared environment offers advantage of facilitating the efficacy of exogenous therapeutic measures on condition that endogenous regeneration has been accelerated.
Modification of Wound Healing Process Using Basic Fibroblast Growth Factor
Authors: Ono Ichiro , Yamashita Toshiharu , Tominaga Akihiro , Akasaka Yoshiharu , Ishii Toshiharu , Jimbow Kowichi
Published: January 2005

In order to identify a means to reduce the scar formation of the skin after incision, this study examined the wound healing effect of bFGF in humans. BFGF was administered at dose of 0.1 and 1 μg per cm of sutured immediately after an operation. The drug was injected once into the dermis of the margins of wounds using a 27 G needle attached to a 1‐ml syringe to the patients. The lengths of the treated wounds varied from 1.5 cm to 23 cm, and the subjects were 2 to 76 years old. Sutured wounds after excision of skin tumors from the face, trunk and limbs and the sutured wounds such as those at the donor sites of full thickness skin grafts were treated with low dose bFGF injections. Postoperative administration of bFGF, inhibited scaring and accelerated healing without any serious side effects. Although double–blind studies are needed, we expect that so‐called scar‐less surgery may be possible by establishing of more sophisticated methods for administering bFGF and its combination with other drugs and/or gene therapy in the future.
Symposium i‐2. pathophysiology and treatment of pressure ulcer Moderated by Takashi Tajiri: Wound Bed Regeneration by Autologous Unfractionated Bone Marrow Transplantation
Authors: Kouraba Sachio , Yasuda Masato , Kumakiri Masanobu , Sakamoto Taisuke , Sugihara Tsuneki
Published: January 2005

Although the pathogenesis of chronic ulcer still remains undefined, one of the characteristics is poor granulation of wound bed, in other words, disruption of wound angiogenesis. Experimental studies have shown that bone marrow (BM) derived endothelial progenitor cells take part in postnatal neovascularivation in cutaneous wound healing. BM cells also contain mesenchymal stem/progenitor cells with pluripotency differentiating into myofibroblasts and fibroblasts. We treated nonhealing leg and foot ulcers sustained over one year by topical transplantation of autologous fresh unfractionated BM‐impregnated collagen sponge (BMiCS). In all patients, the treatments led to rapid generation of well‐vascularized granulation tissue. All of the wounds were healed up completely with conservative treatment or skin grafting. We suggest that direct transplantation of autologous bone marrow cells on the wound may represent a novel procedure for regeneration of disrupted wound healing on recalcitrant cutaneous ulcers.
Viability and Function of Autologous and Allogeneic Fibroblasts Seeded in Dermal Substitutes After Implantation
Authors: Morimoto Naoki , Suzuki Shigehiko , Saso Yasumi , Tomihata Kenji , Taira Tsuguyoshi , Takahashi Yoshitake , Morikawa Noriyuki
Published: January 2005

Aim:  The Aim of this study is to investigate the viability and distribution in vivo of autologous and allogeneic fibroblasts seeded in the collagen sponges after implantation and to clalify which is more effective for wound healing.

Methods:  Three full‐thickness wounds were created on the backs of guinea pigs and acellular substitute, substitute seeded with autologous fibroblasts at the density of 1,0 × 106 cells/cm2, and substitute seeded with allogeneic fibroblasts at the same density were transplanted. Before implantation, fibroblasts were labeled with PKH26.1, 2, 3 weeks after implantation, the epithelized areas were measured and specimens were taken.

Results:  PKH26 labeled autologous and allogeneic fibroblasts remained viabl. In the wound covered with autologous fibroblasts seeded substitute, the epithelization was fastest, and wound contraction was minimal. In contrast, in the wound covered with allogeneic fibroblasts, the epithelization was latest and the contraction was largest

Conclusions:  Allogeneic fibroblasts seeded in the collagen sponge took and remained viable on the grafted area, but adversely affected the wound healing.
Comparison Between Omental Implantation and Omental Patch for Perforated Acetic Acid‐Induced Gastric Ulcer in Rats
Authors: Matoba Yasunori , Tokunaga Akira , Moriyama Yukichi , Iida Shinya , Matsukura Norio , Tajiri Takashi , Katayama Hironori
Published: January 2005

Aim:  Omental implantation, a surgical procedure in which a perforated gastric or duodenal ulcer is repaired by drawing and implanting a portion of the omentum into the digestive tract, accelerates ulcer healing and inhibits ulcer recurrence compared with omental patch from clinical results. To clarify these mechanism and differences, we investigated ulcer healing in two groups.

Methods:  In two groups of rats in which acetic acid‐induced gastric ulcers were perforated. Omental implantation was used for repaired in one group and omental patch was employed in the other group. Basic fibroblast growth factor (bFGF) mRNA‐positive cells were identified and localized by in situ hybridization. Fibroblast growth factor receptor (FGFR)‐positive cells were identified and localized by immunohistochemical analysis.

Results:  Antiinflammatory and angiogenic activity and accelerated collagen synthesis were seen in the omental implantation group. BFGF mRNA‐positive cells (macrophages, fibroblasts, and endothelial cells) and FGFR‐positive cells were seen within the omentum, resulting in abundant collagen production and rapid epithelial regeneration. In the omental patch group, extensive neutrophilic infiltration and ulcer recurrence were seen. Few bFGF mRNA‐positive cells and FGFR‐positive cells were seen within the omentum, resulting to inhibit omentum becoming to be granulation tissue and ulcer healing.

Conclusions:  These results indicated that omental implantation accelerated ulcer healing, and the presence of bFGF mRNA and FGFR played a significant role in this phenomenon.