bluuebunny/crossref_metadata_2025 huggingface dataset:
Interleukin‐1β stimulates macrophage inflammatory protein‐1α and ‐1β expression in human neuronal cells (NT2‐N)
Authors: Guo Chang‐Jiang , Douglas Steven D. , Lai Jian‐Ping , Pleasure David E. , Li Yuan , Williams Marge , Bannerman Peter , Song Li , Ho Wen‐Zhe
Published: March 2003

Chemokines are important mediators in immune responses and inflammatory processes of neuroimmunologic and infectious diseases. Although chemokines are expressed predominantly by cells of the immune system, neurons also express chemokines and chemokine receptors. We report herein that human neuronal cells (NT2‐N) produce macrophage inflammatory protein‐1α and ‐1β (MIP‐1α and MIP‐1β), which could be enhanced by interleukin (IL)‐1β at both mRNA and protein levels. The addition of supernatants from human peripheral blood monocyte‐derived macrophage (MDM) cultures induced MIP‐1β mRNA expression in NT2‐N cells. Anti‐IL‐1β antibody removed most, but not all, of the MDM culture supernatant‐induced MIP‐1β mRNA expression in NT2‐N cells, suggesting that IL‐1β in the MDM culture supernatants is a major factor in the induction of MIP‐1β expression. Investigation of the mechanism(s) responsible for IL‐1β‐induced MIP‐1α and ‐1β expression demonstrated that IL‐1β activated nuclear factor kappa B (NF‐κB) promoter‐directed luciferase activity in NT2‐N cells. Caffeic acid phenethyl ester, a potent and specific inhibitor of activation of NF‐κB, not only blocked IL‐1β‐induced activation of the NF‐κB promoter but also decreased IL‐1β‐induced MIP‐1α and ‐1β expression in NT2‐N cells. These data suggest that NF‐κB is at least partially involved in the IL‐1β‐mediated action on MIP‐1α and ‐1β in NT2‐N cells. IL‐1β‐mediated up‐regulation of β‐chemokine expression may have important implications in the immunopathogenesis of inflammatory diseases in the CNS.
The vocational situation in cases of memory deficits or younger‐onset dementia
Authors: Öhman Annika , Nygård Louise , Borell Lena
Published: March 2003

The aim of the study was to describe how persons with memory deficits or younger‐onset dementia perceived and met their problems, with particular reference to their present and future vocational situation, including describing the parallel observations and attitudes of other significant persons in the vocational context. Data were collected through 16 interviews, one with each of nine subjects and seven respondents from the subjects' work‐places. All interviews were analysed with a comparative method, searching for themes. In addition, subjects' IADL (instrumental activities of daily living) ability and cognition were assessed. The findings demonstrated that the subjects themselves experienced extensive difficulties before their predicament was defined. The difficulties were met by spontaneous and planned strategies on the part of the subjects and by adaptations at the workplaces, but with limited success. The situation also profoundly affected fellow workers. Overall, both the subjects and the work‐place respondents had a positive attitude to continued work as long as the subjects made useful contributions. The possibility of remaining in work seemed to depend on multiple factors, including the individual's apprehension of his or her own ability and the length of the sick‐leave.
Medicate, restrain or seclude? Strategies for dealing with violent and threatening behaviour in a Norwegian university psychiatric hospital
Authors: Wynn Rolf
Published: March 2003

Medicate, restrain or seclude? Strategies for dealing with violent and threatening behaviour in a Norwegian university psychiatric hospital

The aim of this study was to examine staff strategies for applying emergency procedures with patients who were violent or threatening. The study addressed those situations where verbal intervention, voluntary medication, and other interventions had been tried unsuccessfully and where staff found it necessary to apply emergency procedures in the shape of forced emergency medication (pharmacological restraint), physical restraint, and seclusion. The study was conducted in a 100‐bed Norwegian university psychiatric hospital. By retrospectively examining hospital records we found there were 797 episodes of physical restraint, 384 episodes of pharmacological restraint, and 88 episodes of seclusion during a five and a half year period. The preferred emergency procedures varied significantly with patients' sex, age, and diagnoses. Physical restraint was preferred more often with male, younger, and nonpsychotic patients. Pharmacological restraint was preferred more often with female patients and older patients with a nonorganic psychotic disorder. Seclusion was preferred more often with older male patients with an organic psychotic disorder.
Intersimple sequence repeat (ISSR) polymorphisms as a genetic marker system in cotton
Authors: Liu Bao , Wendel Jonathan F.
Published: March 2003

We studied the applicability of intersimple sequence repeat (ISSR) polymorphism in cotton. We found that: (i) the resolving power of agarose gels is poor relative to that provided by sequencing gels; (ii) fluorescent labelling of ISSR amplification primers produced numerous scorable bands; (iii) primer mixing (double priming) generated more bands than the sum of fragments resulting from two single primers, although an unexplained disappearance of several larger fragments also reproducibly occurred; (iv) ISSR fingerprinting patterns are highly heritable; and (v) double priming ISSR is an easy and informative genetic marker system in cotton for revealing both inter‐ and intraspecific variations.
Isolation and characterization of polymorphic microsatellite markers in the blowflies Lucilia illustris and Lucilia sericata
Authors: Florin A.‐B. , Gyllenstrand N.
Published: March 2003

Thirteen polymorphic microsatellite loci were developed for blowflies for use in studies of genetic differentiation in wild populations of Lucilia illustris, to detect the possible occurrence of bottlenecks and to study changes in genetic variation in laboratory populations of Lucilia sericata following artificial bottlenecks. In this preliminary study it was revealed that heterozygosity was lower than expected in wild populations and genetic variation had been lost in the laboratory population despite being kept at a large size.
St Lucia Whiptail Lizard Cnemidophorus vanzoi (Sauria: Teiidae) microsatellite primers
Authors: Rowe Graham , Dickinson Hannah C. , Gibson Richard , Funk Stephan M. , Fa John E.
Published: March 2003

Eight polymorphic microsatellite loci have been characterized for the endangered St Lucia Whiptail Lizard Cnemidophorus vanzoi. Endemic to two small islets, Maria Major and Maria Minor, off the coast of St Lucia, Lesser Antilles, the world population is estimated at < 1000 individuals. However, representatives of the systematically complex genus Cnemidophorus, containing sexual species and parthenogenetic allopolyploids, are distributed widely in North, Central and South America, and on several islands in the Caribbean Sea. These microsatellite markers are being used to monitor the genetic structure of a population of St Lucia Whiptails, recently founded through translocation, on the nearby Praslin Island.
Estimation of gene frequency and heterozygosity from pooled samples
Authors: Ritland K.
Published: March 2003

Pooling of DNA samples can significantly reduce the effort of population studies with DNA markers. I present a statistical model and numerical method for estimating gene frequency when pooled DNA is assayed for the presence/absence of alleles. Analytical and Monte‐Carlo methods examined estimation variance and bias, and hence optimal pool size, under a triangular allele frequency distribution. For gene frequency of rarer alleles, the optimal number of pooled individuals is approximately the inverse of the gene frequency. For heterozygosity, the optimal pool is approximately half the allele number; this results in pools containing, on average, 60% of possible alleles.
Isolation and characterization of microsatellite loci in wild and domestic turkeys (Meleagris gallopavo)
Authors: Latch Emily K. , Smith Edward J. , Rhodes Olin E.
Published: March 2003

We describe the isolation, development and application of seven microsatellite loci in the eastern wild turkey, Meleagris gallopavo silvestris, as well as their amplification and levels of polymorphism in the domestic turkey. The number of alleles per locus ranged from 5 to 15 and average heterozygosity was high for almost all loci. Domestic turkeys showed significantly reduced numbers of alleles per locus and overall heterozygosities when compared to eastern wild turkeys. The high variability in these markers should provide the level of resolution required to continue studies of wild turkey population genetics.
Characterization of 14 microsatellite marker loci in the grey bamboo lemur (Hapalemur griseus)
Authors: Sommer J. A. , Barber R. C. , Huebinger R. M. , Grassi C. , Williamson J. E. , Louis E. E.
Published: March 2003

Fourteen nuclear microsatellite loci were isolated from a genomic DNA library derived from a free‐ranging bamboo lemur (Hapalemur griseus griseus) from Ranomafana National Park, Madagascar. Expected and observed heterozygosity levels were determined for populations of bamboo lemurs from Ranomafana National Park and a nonprotected area near Tsinjoarivo, Madagascar. Although genotype frequencies for all but one marker were within Hardy–Weinberg expectations for the Tsinjoarivo population, this was not the case for the Ranomafana samples. It is probable that the deviation of genotype frequencies from Hardy–Weinberg expectations in Ranomafana was precipitated by the presence of two taxa within the sample population.
Isolation and characterization of highly polymorphic microsatellite loci in the 2‐Spot Ladybird, Adalia bipunctata
Authors: Haddrill P. R. , Majerus M. E. N. , Mayes S.
Published: March 2003

Contrary to theoretical predictions, female 2‐spot ladybirds (Adalia bipunctata) mate many more times than necessary to maintain high fertilisation success and may gain through the acquisition of material or genetic benefits. In order to investigate this mating system in detail, microsatellite markers have been isolated using a modified enrichment technique. Thirty‐nine loci were successfully amplified by polymerase chain reaction (PCR), of which only two were monomorphic. Detailed characterization of ten loci revealed very high levels of polymorphism. These markers are likely to be invaluable tools with which to study population genetics and patterns of paternity in this species.
Novel microsatellite markers for Atlantic sturgeon (Acipenser oxyrinchus) population delineation and broodstock management
Authors: Henderson‐Arzapalo A. , King T. L.
Published: March 2003

Twenty‐one new disomic, polymorphic microsatellite DNA markers were isolated in Atlantic sturgeon, Acipenser oxyrinchus oxyrinchus. These markers yielded a total of 220 alleles in a survey of 16 fish; two to 21 alleles/locus were observed. Each locus segregated in a Mendelian fashion when tested in a family, and a set of 14 of the loci distinguished between siblings and half‐siblings. Average observed heterozygosity ranged from 18.8 to 100.0%, and no linkage disequilibrium was detected. These loci should detect sufficient genetic diversity to allow kinship analysis for broodstock management, gene marking for stocking assessment and life history studies, and delineation of fine‐scale population structure.
A set of 35 consensus primer pairs amplifying genes and introns of plant mitochondrial DNA
Authors: Duminil J. , Pemonge M.‐H. , Petit R. J.
Published: March 2003

Being generally uniparentally inherited, the plant mitochondrial genome is a source of original markers potentially useful for studies of phylogeny and population genetics. We designed 24 new pairs of plant mitochondrial DNA primers that allow amplification of either introns, intergenic regions or genes. They have been defined for consensus over angiosperms and were tested along with 11 previously described mitochondrial primer pairs on 28 plant species representing 19 families of higher plants. The total set allows amplification of 40 kb (∼11%) of the mtDNA genome of Arabidopsis thaliana. The amplification rate ranged between 76% and 100% depending on the species.
Species‐specific TaqMan probes for simultaneous identification of (Gadus morhua L.), haddock (Melanogrammus aeglefinus L.) and whiting (Merlangius merlangus L.).
Authors: Taylor Martin I. , Fox Clive , Rico Ines , Rico Ciro
Published: March 2003

We report the development of a multiplex, single tube (TaqMan) PCR assay for the identification of three commercially important gadoid species, the cod (Gadus morhua L.), the haddock (Melanogrammus aeglefinus L.) and the whiting (Merlangius merlangus L.). The assay unambiguously identifies known adult tissue samples, and ethanol‐preserved eggs from all three species with greater than 98% accuracy, providing a powerful tool for the development of catch independent stock assessment methods, mapping of spawning sites and the detection of commercial fraud in the fishing and food production industries.
Characterization of microsatellite loci in the Jamaican fruit‐eating bat Artibeus jamaicensis and cross‐species amplification
Authors: Ortega J. , Maldonado J. E. , Arita H. T. , Wilkinson G. S. , Fleischer R. C.
Published: March 2003

Artibeus jamaicensis is one of the most common bat species in the neotropics, with a well‐defined polygynous social structure in caves. In order to study behaviour and to examine patterns of paternity and relatedness between different harem groups, we developed 14 microsatellite loci from two different enriched genomic libraries. We screened 125 individuals from two different bat colonies and found that polymorphism ranged from five to 13 alleles. Heterozygosity ranged from 63 to 95%. The primers amplified across 14 bat species, indicating their potential utility for population‐level studies in several closely related bat species.
Tetranucleotide microsatellite DNA loci from the dollar sunfish (Lepomis marginatus)
Authors: Schable Nancy A. , Fischer Robert U. , Glenn Travis C.
Published: March 2003

We describe polymerase chain reaction (PCR) primers and conditions to amplify one dinucleotide and eight tetranucleotide microsatellite DNA loci isolated from the dollar sunfish (Lepomis marginatus). The PCR primers were tested on 20 or more individuals from five populations. The dollar sunfish microsatellite primers developed yielded a high number of alleles (4 to 14 per locus), and high observed heterozygosities (0.500–0.857).
Isolation and characterization of polymorphic microsatellite loci in Acanthoscelides obvelatus Bridwell (Coleoptera: Bruchidae)
Authors: Alvarez N. , Aebi A. , Risterucci A.‐M. , Hossaert‐Mckey M. , Benrey B.
Published: March 2003

Five microsatellite loci were isolated from the bruchid Acanthoscelides obvelatus Bridwell (Coleoptera: Bruchidae). Each locus was polymorphic, with the number of alleles ranging from two to 15. We found high levels of within‐population variation at most loci, with heterozygosity ranging from 0.182 to 0.900. Cross‐species amplification of these loci was tested in two other species of the genus Acanthoscelides, A. obtectus Say and A. argillaceus Sharp.
Polymorphic microsatellites in two different species of the genus Zoogoneticus Meek, 1902 (Goodeidae, Actynopterygii)
Authors: Boto L. , Doadrio I.
Published: March 2003

This report describes the characterization of 13 microsatellites in Zoogoneticus tequila, a goodeid fish that is extinct in nature and exists only in captivity. We also present a preliminary survey of the variability of nine of these microsatellites in both Z. tequila and the congeneric wild species Z. quitzeoensis. Variability was found to be low in Z. tequila (three polymorphic loci with two alleles each), and highest in Z. quitzeoensis (five polymorphic loci with a mean of 4.6 alleles). These genetic markers have applications in the study and management of both species.
Characterization of microsatellite markers in Fagus sylvatica L. and Fagus orientalis Lipsky
Authors: Pastorelli R. , Smulders M. J. M. , VAN’T Westende W. P. C. , Vosman B. , Giannini R. , Vettori C. , Vendramin G. G.
Published: March 2003

Using an enrichment procedure, we cloned microsatellite repeats from European beech (Fagus sylvatica L.) and developed primers for the amplification of microsatellite markers. Six polymorphic loci were characterized which produced 3–21 alleles in 70 individuals from one Italian population, with an observed heterozygosity between 0.58 and 0.85. All six loci amplified fragments which were polymorphic in the closely related species, Fagus orientalis, also. Owing to their very high degree of variation, these markers should be very useful in gene flow studies of these species.
Variation in the growth and survival of Golden Plover Pluvialis apricaria chicks
Authors: Pearce‐Higgins J. W. , Yalden D. W.
Published: March 2003

The growth of Golden Plover Pluvialis apricaria chicks is modelled in detail for the first time. The pattern of growth is typical of postnatal development in waders, although the mean fledging time of 37 days is slower than would be expected from adult body weight. Bill length and weight at hatching had significant effects on the rate of weight gain shown by 2‐day‐old chicks, although this effect was not noticeable at 4 days of age. Chick survival was significantly affected by bill length, as a result of the variation in weight gain. The growth of older chicks was positively correlated with mean minimum temperature. The slow rate of growth exhibited by Golden Plover chicks is discussed in relation to breeding habitat and the effects of weather and hatchling biometrics.
European CuckooCuculus canorusparasitism and host's rejection behaviour in a heavily parasitized Great Reed WarblerAcrocephalus arundinaceuspopulation
Authors: Moskát Csaba , Honza Marcel
Published: March 2003

An unusually high frequency (64%) of European CuckooCuculus canorusparasitism was found in Great Reed WarblerAcrocephalus arundinaceusclutches in central Hungary. Sixty‐four per cent of the parasitized clutches contained one Cuckoo egg, 23% contained two, 10% had three and 3% had four. This means that 58% of the Cuckoo eggs were found in multiply parasitized clutches. In multiple parasitism the laying second Cuckoo removed an egg from the clutch randomly, so preferred neither the host eggs, nor the concurrent Cuckoo egg. Host response towards the parasitic eggs showed 66% acceptance, 12% ejection, 20% desertion and 2% egg burial. We found great variation in both the host and the parasitic egg colour and pattern. This reduces the chance that the parasitic egg's appearance matched that of the hosts’ but, in spite of this, almost perfect mimesis was found in 28% of the Cuckoo eggs. Poorly mimetic Cuckoo eggs were more frequently rejected by Great Reed Warblers than parasite eggs that were very similar to the host eggs. This high level of mimicry sometimes makes it difficult for the observer to identify the parasitic egg, especially when it is similar in size to the host eggs. It is also difficult for the host, as shown by the relatively high recognition error and ejection cost.
Guidelines for assigning species rank†
Authors: Helbig Andreas J. , Knox Alan G. , Parkin David T. , Sangster George , Collinson Martin
Published: March 2003

Developments in several fields of study (including bio‐acoustics and the analysis of DNA) together with reappraisals of the nature of species have impacted significantly on avian taxonomy. The BOU's Taxonomic Subcommittee has developed guidelines for the application of species limits to sympatric, parapatric, allopatric and hybridizing taxa. These are published here to assist researchers understand the rationale behind the committee's taxonomic recommendations relating to the British List.
Mitochondrial DNA diversification among the subspecies of the Silver and Kalij Pheasants, Lophura nycthemera and L. leucomelanos, Phasianidae
Authors: Moulin Sibyle , Randi Ettore , Tabarroni Cristiano , Hennache Alain
Published: March 2003

The taxonomic status of the pheasant superspecies Lophura leucomelanos and Lophura nycthemera has been unclear since 1948. Molecular techniques provided the opportunity to clarify the situation. Using sequences of mitochondrial DNA (800 nucleotides from the D‐loop, plus 400 from the cyt b) from 49 specimens belonging to 10 subspecies (plus two outgroups), we constructed a phylogeny of the subspecies of L. nycthemera and L. leucomelanos. Our data support the monophyly of both species. L. l. lineata and L. l. crawfurdi belong to L. leucomelanos and not to L. nycthemera (suggested by other authors). Our data also confirm a northern locality of origin (Central Buthan) for L. l. moffitti, and have clarified the relationships between subspecies within each species: there are three groups within L. leucomelanos and two within L. nycthemera.
The repeatability and accuracy of axial length and anterior chamber depth measurements from the IOLMaster™*
Authors: Lam Andrew K. C. , Chan Rufina , Pang Peter C. K.
Published: March 2003

Background: Axial length and anterior chamber depth have been measured clinically using conventional ultrasound biometry. Recently, a non‐contact device has become available to measure these parameters. This study evaluated the repeatability and accuracy of this device.

Methods: The axial length and anterior chamber depth were measured by two practitioners on a group of young subjects using the IOLMaster™ followed by a conventional ultrasound biometer operated by a third practitioner. The accommodation was controlled in ultrasound biometry through a full correction on the non‐measured eye and a distant fixation target.

Results: There was good repeatability and accuracy of axial length assessment. The mean difference between the IOLMaster™ and ultrasound biometry was −0.099 , with 95% limits of agreement between 0.66 and −0.85 . The axial length was slightly shorter from the IOLMaster™ and the difference was not significant. The anterior chamber depth was repeatable but was shown to be deeper than the ultrasound results. The mean difference in anterior chamber depth between the IOLMaster™ and ultrasound biometry was 0.15 , with 95% limits of agreement between 0.34 and −0.03 . It is suggested that the former device is not measuring the axial anterior chamber depth.

Conclusions: The IOLMaster™ is a non‐contact ‘optical’ A‐scan which is simple to use and good for axial length assessment. The anterior chamber depth assessment should be further evaluated.
Cooling strategies for patients with severe cerebral insult in ICU (Part 2)
Authors: Price Theresa , McGloin Sarah , Izzard Julie , Gilchrist Mollie
Published: March 2003

Critically ill patients who have sustained a severe cerebral insult will be actively cooled should they develop an elevated body core temperature

Patients who require therapeutic hypothermia for neuroprotection may require the same cooling strategies

A literature review suggested limited evidence to support cooling strategies currently used within one intensive care unit

An experimental approach was used to examine the effects of paracetamol and four external cooling strategies on patients with severe cerebral insult

It is suggested that paracetamol is effective in reducing body core temperature and that fans may not. However, data obtained from the study of the four external cooling strategies were inconclusive
Effect of Alcohol on Body Weight
Authors: Suter Paolo M.
Published: March 2003

Moderate consumption of alcohol has been lauded as a possible protector of the heart. However, alcohol's effects as a promoter of weight gain and obesity, and subsequently cardiovascular disease risk, have not received much attention. This is likely due to the inconsistencies and confounders associated with both observational and experimental research in this area. Some research has shown, however, that alcohol, even in moderate amounts, can suppress fat oxidation, leading to a positive energy balance. In addition, moderate consumers tend to add alcohol to their typical energy intake. Alcohol provides 7.1 kcal/g and therefore contributes a siginificant amount of calories. Unless this greater energy intake and decreased fat oxidation is compensated for through diet and/or exercise, weight gain and possibly obesity can result. In addition, alcohol enhances the deposition of un‐oxidized fat in the abdominal area, which can also raise cardiovascular risk. Patients for whom weight is a concern should be encouraged to decrease their alcohol intake even below the moderate level, which is generally considered as healthy. Fat intake should also be reduced by one gram for every gram of alcohol consumed.
Exercise and Depression in the Older Adult
Authors: Singh Nalin A. , Fiatarone Singh Maria A.
Published: March 2003

Mounting evidence suggests that habitual physical activity is associated with psychological wellbeing and that exercise is an effective alternative treatment for major depressive illness. The elderly population represents a high‐risk group for depressive symptoms and their associated morbidity and mortality due to a variety of biological, pathological, and psychosocial concomitants of aging. Current pharmacological treatment regimens for depression, although effective, are often under‐prescribed or poorly tolerated in this population, particularly in individuals who are frail, very old, or have comorbid diseases. This article presents a rationale for the prescription of exercise as an alternative treatment in the depressed elderly patient, as well as specific recommendations necessary for the practical implementation of this regimen in clinical care and community settings. The paper also discusses areas where additional clinical trial data are needed to target appropriate patients, refine treatment approaches, and determine long‐term efficacy.
Effect of hyperbaric oxygen on human skin cells in culture and in human dermal and skin equivalents
Authors: Dan Dimitrijevich S. , Paranjape Sulabha , Wilson Judy R. , Gracy Robert W. , Mills John G.
Published: March 2003

A critical stage of cutaneous wound healing is the development and maturation of the epidermis. In the aged, and in certain pathologies, this repair process is compromised due to a variety of deficiencies, one of which is tissue oxygenation. Several phases of wound healing are dependent on adequate tissue oxygen levels, and hyperbaric oxygenation has been shown to transiently elevate these levels. The use of human cell monolayers, dermal equivalents and human skin equivalents provide excellent opportunities for studying wound healing using in vivo relevant models. The goal of this study was to examine the effect of hyperbaric oxygen on cell proliferation, differentiation, and matrix biosynthesis in monolayer cultures and epidermopoiesis in the developing skin equivalent. Normal human dermal fibroblasts, keratinocytes and melanocytes, dermal equivalents and skin equivalents were exposed to hyperbaric oxygen at pressures up to three atmospheres, for up to 10 consecutive daily treatments lasting 90 minutes each. Increase in fibroblast proliferation (cf., 30% at 1 atmosphere after 10 days treatment), was observed without a significant effect on proliferation of normal human melanocytes and glycosaminoglycan synthesis. Stimulation of collagen synthesis after two days of treatment was only significant at 1 atmosphere (about 20% increase) but this differential was not observed after 5 days of treatment. Hyperbaric oxygenation above 2 atmospheres, inhibited proliferation of fibroblasts and keratinocytes in cell monolayer cultures (e.g., a 10 day treatment at 3 atmospheres appeared cytostatic to keratinocytes). In contrast, hyperbaric treatment up to 3 atmospheres dramatically enhanced keratinocyte differentiation, and epidermopoiesis in the complete human skin equivalent. These results support the importance of hyperbaric oxygen therapy in wound healing, and should provide an insight into oxygen utilization during repair of peripheral human tissue. The results also show the utility of the human skin equivalent as a model for evaluation of parameters involved in wound healing. (WOUND REP REG 1999;7:53–64)
Collagen fiber orientation as quantified by small angle light scattering in wounds treated with transforming growth factor‐β2 and its neutalizing antibody
Authors: Bowes Leyda E , Jimenez Maria C , Hiester Erik D , Sacks Michael S , Brahmatewari Just , Mertz Patricia , Eaglstein William H
Published: March 2003

The purpose of this study was determine quantitative differences in collagen fiber orientation in a wound healing model in the presence of transforming growth factor‐β2 and anti‐transforming growth factor‐β2,3 antibody. Full‐thickness wounds were made in the paravertebral area of two young pigs. Wounds were treated once, topically, with either transforming growth factor‐β2 or anti‐transforming growth factor‐β2 antibody, or with methylcellulose gel. Control wounds were left untreated. Tissue biopsies were obtained from each wound on days 7, 14 and 46 post wounding. Tissue sections were stained with hematoxylin and eosin, and collagen fiber preferred orientation was quantified using small angle light scattering. Our results indicated that wounds treated with transforming growth factor‐β2 and anti‐transforming growth factor‐β2,3 antibody had a significantly higher degree of orientation of collagen fibers than normal unwounded skin on days 7, 14 and 46 (p < 0.001). Transforming growth factor‐β2– treated wounds had a higher degree of orientation of collagen fibers than control wounds on days 7 and 14 (p < 0.001), and control wounds displayed a higher degree of orientation than wounds treated with anti‐transforming growth factor‐β2,3 and normal unwounded skin at all time points (p < 0.001). These results suggest that differences in the dermal collagen degree of orientation correlate with scarring, and show that small angle light scattering can be used quantitatively to assess differences in the collagen fiber architecture of dermal wounds.
Evaluation of clinically applicable exsanguination treatments to alleviate venous congestion in an animal skin flap model
Authors: Cottler Patrick S , Gampper Thomas J , Rodeheaver George T , Skalak Thomas C
Published: March 2003

This study compares the effectiveness of alleviating venous congestion with mechanically‐made outlets or leech therapy in promoting skin flap survival. Free flaps of abdominal skin (3 × 6 cm) were raised on Sprague‐Dawley rats and subjected to ischemic events, simulating venous congestion. Animals received 1) no treatment; 2) two treatments involving two 18‐gauge needle‐puncture outlets; or 3) two sessions of leech therapy. Flap perfusion was monitored with a scanning laser Doppler flowmeter. Photographic images of flaps at 7 days were assessed for areas of normal tissue (n = 15), and laser Doppler flowmeter data consisted of control (n = 6), outlet (n = 6), and leech (n = 7). Both the needle‐puncture outlet (40.0%± 9.24%) and leech treated (34.6%± 7.34%) groups had a significantly greater surviving skin area than untreated control flaps (8.0%± 5.0%), with 2 of 15 flaps receiving mechanical outlets exhibiting > 90% surviving area. After 7 days, laser Doppler flowmeter data showed greater mean perfusion in the outlet (71.7%± 16.8%) and leech (92.6%± 17.2%) treated groups, compared to controls (15.2%± 10.2%). There was a significant increase in perfusion in the outlet (13.3%± 6.2%) and leech (9.1%± 1.1%) treated groups from the end of secondary ischemia to day 7 (p < 0.05) compared to controls. The results suggest that two spatially separated outlets are as effective as one leech in increasing the area of surviving skin in venous congested flaps.
Comparative trial of octyl‐cyanoacrylate and silver sulfadiazine for the treatment of full‐thickness skin wounds
Authors: Singer Adam J , Berrutti Luis , Mcclain Steve A
Published: March 2003

A prospective, randomized, blinded, controlled experimental trial was performed in pigs to compare the rates of reepithelialization of 126 full‐thickness cutaneous 4‐mm punches treated with an octyl‐cyanoacrylate spray, silver sulfadiazine, or a dry gauze (controls). Full thickness biopsies were taken 7, 14, or 30 days later for histopathological evaluation of hematoxylin and eosin stained tissue sections by a dermatopathologist. The primary outcome measure was the proportion of wounds completely re‐epithelialized at days 7 and 14. Secondary outcomes were the rates of infection, foreign body reactions, and the depth of any resulting cutaneous dells measured with a micrometer. Between‐group comparisons were performed with ANOVA or Chi‐square tests. Octyl‐cyanoacrylate treated wounds re‐epithelialized more slowly, as fewer wounds treated with octyl‐cyanoacrylate were re‐epithelialized at day 7 in comparison with silver sulfadiazine or control wounds (50% vs. 90% vs. 100%, p < 0.001). There were no infections or foreign body type reactions. Amounts of granulation tissue were similar among groups. Octyl‐cyanoacrylate wounds were more depressed than silver sulfadiazine wounds at days 7 and 14 yet had similar histopathological characteristics at day 30. We conclude that treatment of small, full thickness cutaneous wounds with octyl‐cyanoacrylate results in delayed re‐epithelialization and dermal repair in comparison with silver sulfadiazine, yet it does not result in any foreign body–type reaction. However, by 30 days, histopathological wound characteristics were similar in all groups.
Denervation retards but does not prevent toetip regeneration
Authors: Mohammad Khalid S , Neufeld Daniel A
Published: March 2003

Toetips of mammals regrow after amputation by a process similar, but not identical, to that which occurs during regeneration of a newt limb. Nerve is needed as a mitotic stimulant for newt limb regeneration but the requirement for nerve during rodent digit‐tip regeneration is not known. Nerve dependence in rats was tested by severing the sciatic nerve in one hindlimb, amputating digit‐tips from the central digits of both hind feet, and comparing the amount of regrowth in innervated and denervated digits. Denervation delayed soft‐tissue wound healing. However, denervation did not significantly affect bone regrowth when animals were examined at one month. Because we suspected delayed bone regrowth, we used a new method that we developed to follow bone growth at several time points in each animal. Termed visible bone fluorescence through nail, this technique used serial injections of fluorescent calcium‐deposition markers and observation through the toenails to observe bone growth in living animals. Using this method it was possible to detect retarded bone regrowth in denervated digits. Thus, although denervation of rodent tips delayed both soft tissue healing and bone regrowth, it did not prevent ultimate restitution of the amputated part. This suggests that neurotrophic stimulation in the mammalian digit‐tip is not identical to that documented during newt limb regeneration, and that growth stimulation may be provided by tissues other than nerve.
Gene therapy in wound repair and regeneration
Authors: Yao Feng , Eriksson Elof
Published: March 2003

The potential use of gene therapy to treat human disease increases with the development of various physical, chemical, and biological methods to deliver genes to mammalian cells, and with our rapidly expanding knowledge of the human genome. One area of therapeutic interest for gene therapy is the treatment of wound healing disorders. Most recently, recombinant human growth factor therapy has been examined as a means to treat problem wounds. However, this approach suffers from the difficulty in providing an accurate dose of growth factor and the expense of the recombinant proteins. Delivery of a gene that could be expressed within the wound is an attractive alternative to application of the protein. This review discusses several methods that have been used to deliver genes encoding growth factor proteins into wounds and the advantages/disadvantages of each approach. Novel methods to regulate the expression of the transgene are also presented, highlighting the ability of these unique vector systems to adjust gene dose as the wound heals. We expect that gene therapy will become a significant treatment modality for those wound healing pathologies refractory to other wound management approaches in the years ahead.
DEVELOPMENT OF NONINVASIVE AND QUANTITATIVE METHODOLOGIES FOR THE ASSESSMENT OF CHRONIC ULCERS AND SCARS IN HUMANS
Authors: Timar‐Banu Olimpia , Beauregard Hughes , Tousignant Jacqueline , Lassonde Michel , Harris Phillip , Viau Gilles , Vachon Luc , Levy Emile , Abribat Thierry
Published: March 2003

Chronic ulcers are a significant and common cause of morbidity and mortality worldwide. They disrupt the epidermis and dermis, resulting in a loss of barrier function. Keloids and hypertrophic scars (benign cutaneous tumors) represent an abnormal healing response. These fibroproliferative disorders are characterized by an overabundance of collagen and accumulation of extracellular matrix due to an imbalance between synthesis and degradation, culminating in excessive scarring. The objectives of this study were to evaluate and compare noninvasive biophysical methods for the measurement of outstanding quantitative parameters of scars and chronic ulcers, and to establish correlations between the parameters measured and the results of conventional subjective clinical evaluations. The development of new technologies, based on ultrasonography and laser Doppler, makes possible new dermatological evaluation methods. Fifteen patients (6 females and 9 males) with 15 chronic ulcers (4 diabetic ulcers, 10 venous ulcers and 1 pressure ulcer) and 30 patients (19 females and 11 males) with 30 scars (25 hypertrophic and 5 keloids) were included in this study. Clinical evaluation was performed by a dermatologist, an aesthetic surgeon and an endocrinologist. Biophysical measurements were used to assess local blood flow by laser Doppler flowmetry (Moor DRT4), thickness and echogenicity by high frequency ultrasonography (20 MHz, Dermascan C) and ulcer linear dimensions by image analysis. Our results show that blood flow within the ulcers and scars was higher than within normal skin. Also, skin thickness of chronic ulcers was decreased when compared to normal skin; the thickness of hypertrophic scars, but not of keloids, was increased in comparison to normal skin, and presented the possibility of measuring wound and scar surfaces with precision. In summary, this pilot study established the feasibility of measuring various biophysical parameters and adapted their potential utility to research on wounds.
Dermatitis during radiation for vulvar carcinoma: prevention and treatment with granulocyte‐macrophage colony‐stimulating factor impregnated gauze
Authors: Kouvaris John R. , Kouloulias Vassilios E. , Plataniotis George A. , Balafouta Ersi J. , Vlahos Lampros J.
Published: March 2003

The aim of this study was to determine the effectiveness of granulocyte‐macrophage colony‐stimulating factor (GM‐CSF) impregnated gauze in preventing or healing radiation‐induced dermatitis. Sixty‐one patients were irradiated for vulvar carcinoma. Thirty‐seven applied steroid cream at irradiated areas throughout radiotherapy (Group A) and 24 patients applied additionally GM‐CSF impregnated gauze (40μg/cm2 of skin‐irradiated area, twice per day) in addition to the steroid cream, after 20Gy of irradiation (Group B). The score of skin reactions (P=0.008, chi2 test) and the time interval of radiotherapy interruption (P=0.037, Mann‐Whitney U test) were statistically significantly reduced in Group B patients. Multivariate analysis of variance showed for this group not only a significant reduction in the Sum of Gross Dermatitis Scoring (P<0.001, adjusted for Duration of Dermatitis) but also a significant reduction of the healing time (P=0.02, adjusted for Sum of Gross Dermatitis Scoring). The pain grading was less (P=0.014, chi2 test) and pain reduction was noticed sooner after the application of GM‐CSF impregnated gauze (P=0.0017, Mann‐Whitney U test). Multivariate logistic regression analysis showed that the only significant effect on dermatitis score is due to Body Mass Index (P=0.034) and the application of GM‐CSF (P=0.008). GM‐CSF impregnated gauze can be effective in preventing and healing radiation‐induced dermatitis and in reducing the interruption intervals of radiotherapy for vulvar carcinomas.
Effects of an aqueous extract from the leaves of Chromolaena odorata (Eupolin) on the proliferation of human keratinocytes and on their migration in an in vitro model of reepithelialization
Authors: Phan Thang T. , Hughes Margaret A. , Cherry George W.
Published: March 2003

Eupolin ointment, prepared from the leaves of Chromolaena odorata, has been shown to promote the healing of soft tissue wounds and burns in Vietnam. However, the mechanism by which this agent affects cells involved in the wound healing process is unknown. Cultured human keratinocytes were used in this study to investigate the effects of the Eupolin extract in vitro on processes involved in wound reepithelialization. Keratinocyte proliferation was monitored by a colorimetric assay and migration by the closure of a denuded area scratched in a confluent monolayer. Human keratinocyte proliferation was stimulated by low concentrations of the extract (from 0.1 to 5 μg/ml), cell differentiation by higher concentrations (50 to 300 μg/ml), and migration by intermediate concentrations (5 to 60 μg/ml). The increased proliferation and migration of human keratinocytes observed in vitro might explain, in part, the beneficial effects that have been observed in the clinic.
Retroviral delivery of dominant‐negative vascular endothelial growth factor receptor type 2 to murine wounds inhibits wound angiogenesis
Authors: Tsou Raymond , Fathke Carrie , Wilson Lynne , Wallace Ken , Gibran Nicole , Isik Frank
Published: March 2003

Vascular endothelial growth factor (VEGF) is a potent paracrine signal for initiating angiogenesis. Although VEGF can bind to several cell surface receptors, VEGF receptor type 2 (VEGFR2, a.k.a. KDR or flk‐1) is the primary receptor responsible for VEGF‐induced endothelial cell proliferation. To determine whether the VEGF‐VEGFR2 signaling axis has an important role in wound healing angiogenesis, we used a retrovirus to deliver a signaling‐defective truncated VEGFR2 (tm VEGFR2) to block VEGF‐VEGFR2–induced endothelial cell proliferation in murine wounds. We show that the retroviral construct effectively blocked phosphorylation of VEGFR2 in vitro and we were able to express the truncated receptor in murine wounds. We achieved significant reduction of angiogenesis and granulation tissue formation in murine wounds, but this did not lead to delayed wound closure. In contrast, there was a corresponding increase in wound contraction, showing that functional VEGFR2 intracellular signaling is not critical for normal closure of excisional dermal wounds. Our results show a novel relationship between wound bed vascularity and wound contraction. (WOUND REP REG 2002;10:222–229)
Glucan stimulates human dermal fibroblast collagen biosynthesis through a nuclear factor‐1 dependent mechanism
Authors: Wei Duo , Zhang Leiying , Williams David L. , Browder I. William
Published: March 2003

Glucan, an immunomodulator, has been reported to increase collagen deposition and tensile strength in experimental models of wound repair. Previous data suggest that glucan modulates wound healing via an indirect mechanism in which macrophages are stimulated to release growth factors and cytokines. However, recent data have shown the presence of glucan receptors on normal human dermal fibroblasts, suggesting that glucans may be able to directly stimulate fibroblast collagen biosynthesis. To test this hypothesis, we examined the effect of glucan on collagen biosynthesis in normal human dermal fibroblasts. We assessed nuclear factor‐1 (NF‐1) activation, procollagen mRNA expression, collagen biosynthesis, and whether there was a causal link between glucan treatment, NF‐1 activation, and collagen expression. Glucan (1 µg/ml) increased NF‐1 binding activity by 46% (8 hours), 64% (24 hours), 215% (36 hours), and 119% (48 hours) in cultured normal human dermal fibroblasts. Alpha 1(I) and α1 (III) procollagen mRNA were increased in glucan‐treated normal human dermal fibroblasts when compared with the untreated fibroblasts. Collagen synthesis was increased at 24 hours and 48 hours following glucan treatment of normal human dermal fibroblasts. Down‐regulation of NF‐1 by pentifylline inhibited glucan‐induced procollagen mRNA expression. These data indicate that glucan can directly stimulate human fibroblast collagen biosynthesis through an NF‐1–dependent mechanism. ( WOUND REP REG 2002;10:168)
Potential role of anaerobic cocci in impaired human wound healing
Authors: Wall Ivan B. , Davies Charlotte E. , Hill Katja E. , Wilson Melanie J. , Stephens Phil , Harding Keith G. , Thomas David W.
Published: March 2003

Although more than 80% of infected and 70% of noninfected leg ulcers have been shown to harbor anaerobic organisms, their role in mediating impaired wound healing in the skin is frequently overlooked. There is now increasing evidence that the gram‐positive anaerobic cocci play a role (both directly and indirectly) in mediating impaired wound healing in vivo. This article discusses the mechanisms by which these microorganisms may interfere with the inflammation, repair, and remodeling phases of the wound healing process. (WOUND REP REG 2002;10:–353)
Transformation of Fe–Ti gabbro to coronite, eclogite and amphibolite in the Baie du Nord segment, Manicouagan Imbricate Zone, eastern Grenville Province
Authors: Cox  , Indares
Published: March 2003

Fe–Ti gabbros from the Baie du Nord Segment of the Manicouagan Imbricate Zone, metamorphosed under high P–T  conditions during the Grenvillian orogeny, have been the focus of a detailed micropetrological study. Textures and mineral chemistry suggest that the mineral assemblages represent progressive stages of metamorphic transformation resulting in the formation of coronas, pseudomorphs after igneous phases (transitional) and true, granoblastic eclogites. The transitional and eclogitic samples also have coronas which are developed locally around igneous xenocrysts of plagioclase and olivine. The deformed margins of coronitic Fe–Ti gabbros are transformed to amphibolite and contain clinopyroxene‐bearing leucosomes with garnet poikiloblasts that are indicative of high‐P–T  dehydration melting. Interpretation of garnet zoning and thermobarometry suggest that the highest P–T conditions are recorded by coronas around xenocrysts (c. 720–800 °C at 14–17 kbar) and garnet–clinopyroxene cores in granoblastic assemblages (c. 740–820 °C at 13–17 kbar) in the eclogitic samples. Re‐equilibration during the early stages of exhumation at high‐T  conditions (>700 °C) affected all samples, and is evidenced by the widespread development of pargasite‐bearing plagioclase collars in the coronitic and transitional metagabbros and by widespread re‐equilibration of the eclogites giving lower P–T  estimates at grain boundaries. However, the difference in calculated pressure conditions between coronite and eclogite samples is consistent with increasing pressure (depth) from the coronites (11–13 kbar) to the eclogites (13–17 kbar). The P–T  conditions recorded by these rocks define a metamorphic field gradient which suggests high heat flow through the lower crust during the Grenvillian orogeny.