bluuebunny/crossref_metadata_2025 huggingface dataset:
Third Ways in Europe?
Authors: Bonoli Giuliano , Powell Martin
Published: July 2002

It has been claimed that there is a global Third Way (TW) debate. Giddens (2001: 1) writes that, ‘Across the world left of centre governments are attempting to institute third way programmes – whether or not they favour the term itself. ‘ He claims that there are self-declared third way parties in power in the UK, New Zealand, Korea, Taiwan, Brazil, Argentina and Chile, among many other countries. Similarly, according to Blair (2001), the ideas associated with the TW are still the wave of the future for progressive politics. From Latin America to Europe to parts of Asia, TW politics or ‘progressive government’ is exerting a huge influence on global politics. The TW is seen as a trailblazer for a new global social policy, a new model for a new millennium (e.g. McGuire, 1998/9). One of the main blueprints for the new politics (Giddens, 1998) has been translated into many languages. A number of international meetings in Paris and Florence have discussed the TW. British Prime Minister Tony Blair and German Chancellor Gerhard Schröder issued a joint paper, ‘The Third Way/Die Neue Mitte’ (Blair and Schröder, 1999) that was drafted by Peter Mandelson and Bodo Hombach. Hombach's book has been translated into English as ‘The New Centre’ (Hombach, 2000), with a preface by Tony Giddens and an introduction by Mark Leonard.
Local Youth Projects: Political Rhetoric and Strategies. The Swedish Case
Authors: Lundahl Lisbeth
Published: July 2002

In Sweden, youth matters increasingly form a political field of their own. Youth politics may be related to governance and policy research on the one hand, and research on young people's lives on the other. Both have been influenced by similar questions, concerning the freedom of actors and the power of structures. Research specifically on youth politics is however scanty, in Sweden and internationally. The article analyses recent Swedish youth politics, based on interviews with local politicians and officials. Questions posed are: Which youth matters are prioritised at the local political agenda, and why? How is the division of responsibilities in youth matters between different actors perceived? Which importance and functions are attributed to local youth projects? To what extent do recent youth politics mirror late-modern assumptions of self-governance, individual choice and risk-taking? To what extent do they reflect assumptions of an earlier dominant social welfare paradigm? It is concluded that local youth politics in general contain elements of collectivist welfare thinking, and a neo-liberal or post-modern ideology is barely visible. Nevertheless, flexibility, autonomy and decentralisation assumptions about local authorities and young people become more prominent around youth projects. The municipality is also more likely to be identified as a strategic, risk-taking actor here.
Positive patch tests with a dermatophagoides mix relate to an increased responsiveness to standard patch test allergens
Authors: Brasch Jochen , Uter Wolfgang , Dibo Maher , Stockfleth Eggert , Swensson Ole , Christophers Enno
Published: July 2002

The diagnostic meaningfulness of patch tests with house dust mite allergens is still questionable. Our own impression has been that positive results with a dermatophagoides mix may occur preferentially in patients with a generally enhanced responsiveness to contact allergens. To check this, all of our patients allocated to patch testing with the standard series were additionally patch tested with a dermatophagoides mix by the same technique that was used for standard contact allergens. Out of 571 patients tested, 188 showed delayed responses to this mix that were indistinguishable from typical allergic patch test reactions but of no apparent clinical relevance. No relationship was found between positive dermatophagoides patch tests and an atopic disposition of the patients or characteristics of their eczema. However, 64.4% of the patients with a positive dermatophagoides patch test showed a response to at least 1 contact allergen of the standard series, compared to only 56.4% of the patients without a positive dermatophagoides reaction (p < 0.05). The reactivity to the mite mix was not related to the responsiveness towards any particular contact allergens. We suppose that some unidentified factors may contribute to positive reactions to the dermatophagoides mix that may also favour an enhanced general responsiveness to contact allergens.
Lyral® has been included in the patch test standard series in Germany
Authors: Geier Johannes , Brasch Jochen , Schnuch Axel , Lessmann Holger , Pirker Claudia , Frosch Peter J. ,   for the Information Network of Departments of Dermatology (IVDK)and the German Contact Dermatitis Research Group (DKG)
Published: July 2002

Lyral® 5% pet. was tested in 3245 consecutive patch test patients in 20 departments of dermatology in order (i) to check the diagnostic quality of this patch test preparation, (ii) to examine concomitant reactions to Lyral and fragrance mix (FM), and (iii) to assess the frequency of contact allergy to Lyral in an unselected patch test population of German dermatological clinics. 62 patients reacted to Lyral, i.e. 1.9%. One third of the positive reactions were + + and + + + . The reaction index was 0.27. Thus, the test preparation can be regarded a good diagnostic tool. Lyral and fragrance mix (FM) were tested in parallel in 3185 patients. Of these, 300 (9.4%) reacted to FM, and 59 (1.9%) to Lyral. In 40 patients, positive reactions to both occurred, which is 13.3% of those reacting to FM, and 67.8% of those reacting to Lyral. So the concordance of positive test reactions to Lyral and FM was only slight. Based on these results, the German Contact Dermatitis Research Group (DKG) decided to add Lyral 5% pet. to the standard series.
Potent analgesic and anti‐inflammatory actions of a novel thymulin‐related peptide in the rat
Authors: Safieh‐Garabedian Bared , Dardenne Mireille , Pléau Jean Marie , Saadé Nayef E
Published: July 2002

The present study examines the effect of PAT (peptide analogue of thymulin) in two rat models of inflammatory hyperalgesia induced by either i.pl. (1.25 μg in 50 μl saline) or i.p. (50 μg in 100 μl) injections of endotoxin ET.

Pretreatment with PAT (1, 5 or 25 μg in 100 μl saline, i.p.) decreased, in a dose dependent manner, both mechanical hyperalgesia, determined by the paw pressure (PP) test and thermal hyperalgesia determined by the hot plate (HP), the paw immersion (PI) and the tail flick (TF) tests.

Compared to the tripeptides K(D)PT and K(D)PV, known to antagonize interleukin (IL)‐1β or IL‐1β and PGE2 mechanisms, PAT, at lower dosages, exerted stronger anti‐hyperalgesic effects.

When compared with the effect of a steroidal (dexamethasone) and a non‐steroidal (indomethacin) anti‐inflammatory drugs (NSAID), PAT demonstrated equal analgesic actions.

Pretreatment with PAT, reduced significantly the increased concentration of IL‐1β, IL‐6, TNF‐α and NGF due to i.pl. injection of ET.

Injection of i.p. ET produced sickness behaviour characterized by hyperalgesia and fever. Pretreatment with PAT prevented the hyperalgesia and maintained the body temperature within the normal range and was accompanied by a down‐regulation of the levels of pro‐inflammatory cytokines and PGE2 in the liver.

PAT, in all doses used, did not result in any evident changes in the physiological parameters or in the normal behaviour of the rats.

The anti‐hyperalgesic and anti‐inflammatory effects of PAT can be attributed, at least partially, to the down‐regulation of pro‐inflammatory mediators.

British Journal of Pharmacology (2002) 136, 947–955. doi:10.1038/sj.bjp.0704793
Effect of inositol 1,4,5-trisphosphate receptor stimulation on mitochondrial [Ca2+] and secretion in chromaffin cells
Authors: MONTERO Mayte , ALONSO Maria Teresa , ALBILLOS Almudena , CUCHILLO-IBÁÑEZ Inmaculada , OLIVARES Román , VILLALOBOS Carlos , ALVAREZ Javier
Published: July 2002

Ca2+ uptake by mitochondria is a potentially important buffering system able to control cytosolic [Ca2+]. In chromaffin cells, we have shown previously that stimulation of either Ca2+ entry or Ca2+ release via ryanodine receptors triggers large increases in mitochondrial [Ca2+] ([Ca2+]M) approaching the millimolar range, whose blockade dramatically enhances catecholamine secretion [Montero, Alonso, Carnicero, Cuchillo-Ibañez, Albillos, Garcia, Carcia-Sancho and Alvarez (2000) Nat. Cell Biol. 2, 57–61]. In the present study, we have studied the effect of stimulation of inositol 1,4,5-trisphosphate (InsP3) receptors using histamine. We find that histamine produces a heterogeneous increase in [Ca2+]M, reaching peak levels at approx. 1μM in 70% of the mitochondrial space to several hundred micromolar in 2–3% of mitochondria. Intermediate levels were found in the rest of the mitochondrial space. Single-cell imaging experiments with aequorin showed that the heterogeneity had both an intercellular and a subcellular origin. Those mitochondria responding to histamine with increases in [Ca2+]M much greater than 1μM (30%) were the same as those that also responded with large increases in [Ca2+]M following stimulation with either high-K+ medium or caffeine. Blocking mitochondrial Ca2+ uptake with protonophores or mitochondrial inhibitors also enhanced catecholamine secretion induced by histamine. These results suggest that some InsP3 receptors tightly co-localize with ryanodine receptors and voltage-dependent Ca2+ channels in defined subplasmalemmal functional units designed to control secretion induced by different stimuli.
Assembly of the ribonucleoprotein complex containing the mRNA of the β-subunit of the mitochondrial H+-ATP synthase requires the participation of two distal cis-acting elements and a complex set of cellular trans-acting proteins
Authors: RICART Javier , IZQUIERDO José M. , Di LIEGRO Carlo M. , CUEZVA José M.
Published: July 2002

The mRNA encoding the β-subunit of the mitochondrial H+-ATP synthase (β-F1-ATPase) is localized in an approx. 150nm structure of the hepatocyte of mammals. In the present study, we have investigated the cis- and trans-acting factors involved in the generation of the ribonucleoprotein complex containing β-F1-ATPase mRNA. Two cis-acting elements (β1.2 and 3′β) have been identified. The β1.2 element is placed in the open reading frame, downstream of the region encoding the mitochondrial pre-sequence of the protein. The 3′β element is the 3′ non-translated region of the mRNA. Complex sets of proteins from the soluble and non-soluble fractions of the liver interact with the β1.2 and 3′β elements. A soluble p88, present also in reticulocyte lysate, displays binding specificity for both the cis-acting elements. Sedimentation and high-resolution in situ hybridization experiments showed that the structure containing the rat liver β-F1-ATPase mRNA is found in fractions of high sucrose concentration, where large polysomes sediment. Treatment of liver extracts with EDTA promoted the mobilization of β-F1-ATPase mRNA to fractions of lower sucrose concentration, suggesting that the structure containing β-F1-ATPase mRNA is a large polysome. Finally, in vitro reconstitution experiments with reticulocyte lysate, using either the full-length, mutant or chimaeric versions of β-F1-ATPase mRNA, reveal that the assembly of the β-F1-ATPase mRNA polysome requires the co-operation of both the cis-acting mRNA determinants. The present study illustrates the existence of an intramolecular RNA cross-talking required for the association of the mRNA with the translational machinery.
Genomic organization of the mouse peroxisome proliferator-activated receptor β/δ gene: alternative promoter usage and splicing yield transcripts exhibiting differential translational efficiency
Authors: LARSEN Leif K. , AMRI Ez-Zoubir , MANDRUP Susanne , PACOT Corinne , KRISTIANSEN Karsten
Published: July 2002

Peroxisome proliferator-activated receptor (PPAR) β/Δ is ubiquitously expressed, but the level of expression differs markedly between different cell types. In order to determine the molecular mechanisms governing PPARβ/Δ gene expression, we have isolated and characterized the mouse gene encoding PPARβ/Δ. The gene spans approx. 41 kb and comprises 11 exons of which the six exons located in the 3′-end of the gene are included in all transcripts. Primer-extension and 5′-rapid amplification of cDNA ends experiments revealed the presence of multiple transcription start points and splice variants, originating from the use of at least four different promoters. One of these transcription start points was found to be used predominantly in all tissues examined. Initiation from this major transcription start point gives rise to a transcript with a 548nt 5′-untranslated leader containing eight upstream AUG codons. We show that the presence of the 548nt leader resulted in a low translational efficiency of the corresponding PPARβ/Δ mRNA and propose, based on structural features of the 5′-untranslated region, that translational initiation may be mediated via an internal ribosome entry site-dependent mechanism.
Localization and function of a calmodulin–apocalmodulin-binding domain in the N-terminal part of the type 1 inositol 1,4,5-trisphosphate receptor
Authors: SIENAERT Ilse , KASRI Nael NADIF , VANLINGEN Sara , PARYS Jan B. , CALLEWAERT Geert , MISSIAEN Ludwig , de SMEDT Humbert
Published: July 2002

Calmodulin (CaM) is a ubiquitous protein that plays a critical role in regulating cellular functions by altering the activity of a large number of proteins, including the d-myo-inositol 1,4,5-trisphosphate (IP3) receptor (IP3R). CaM inhibits IP3 binding in both the presence and absence of Ca2+ and IP3-induced Ca2+ release in the presence of Ca2+. We have now mapped and characterized a Ca2+-independent CaM-binding site in the N-terminal part of the type 1 IP3R (IP3R1). This site could be responsible for the inhibitory effects of CaM on IP3 binding. We therefore expressed the N-terminal 581 amino acids of IP3R1 as a His-tagged recombinant protein, containing the functional IP3-binding pocket. We showed that CaM, both in the presence and absence of Ca2+, inhibited IP3 binding to this recombinant protein with an IC50 of approx. 2μM. Deletion of the N-terminal 225 amino acids completely abolished the effects of both Ca2+ and CaM on IP3 binding. We mapped the Ca2+-independent CaM-binding site to a recombinant glutathione S-transferase fusion protein containing the first 159 amino acids of IP3R1 and then made different synthetic peptides overlapping this region. We demonstrated that two synthetic peptides matching amino acids 49–81 and 106–128 bound CaM independently of Ca2+ and could reverse the inhibition of IP3 binding caused by CaM. This suggests that these sequences are components of a discontinuous Ca2+-independent CaM-binding domain, which is probably involved in the inhibition of IP3 binding by CaM.
Decreased activity and enhanced nuclear export of CCAAT-enhancer-binding protein β during inhibition of adipogenesis by ceramide
Authors: SPROTT Kam M. , CHUMLEY Michael J. , HANSON Janean M. , DOBROWSKY Rick T.
Published: July 2002

To identify novel molecular mechanisms by which ceramide regulates cell differentiation, we examined its effect on adipogenesis of 3T3-L1 preadipocytes. Hormonal stimulation of 3T3-L1 preadipocytes induced formation of triacylglycerol-laden adipocytes over 7days; in part, via the co-ordinated action of CCAAT-enhancer-binding proteins α, β and δ (C/EBP-α, -β and -δ) and peroxisome-proliferator-activated receptor γ (PPARγ). The addition of exogenous N-acetylsphingosine (C2-ceramide) or increasing endogenous ceramide levels inhibited the expression of C/EBPα and PPARγ, and blocked adipocyte development. C2-ceramide did not decrease the cellular expression of C/EBPβ, which is required for expression of C/EBPα and PPARγ, but significantly blocked its transcriptional activity from a promoter construct after 24h. The ceramide-induced decrease in the transcriptional activity of C/EBPβ correlated with a strong decrease in its phosphorylation, DNA-binding ability and nuclear localization at 24h. However, ceramide did not change the nuclear level of C/EBPβ after a period of 4 or 16h, suggesting that it was not affecting nuclear import. CRM1 (more recently named ‘exportin-1') is a nuclear membrane protein that regulates protein export from the nucleus by binding to a specific nuclear export sequence. Leptomycin B is an inhibitor of CRM1/exportin-1, and reversed the ceramide-induced decrease in nuclear C/EBPβ at 24h. Taken together, these data support the hypothesis that ceramide may inhibit adipogenesis, at least in part, by enhancing dephosphorylation and premature nuclear export of C/EBPβ at a time when its maximal transcriptional activity is required to drive adipogenesis.
Transcriptional regulation of the human carboxyl ester lipase gene in THP-1 monocytes: an E-box required for activation binds upstream stimulatory factors 1 and 2
Authors: BENGTSSON Sara H.M. , MADEYSKI-BENGTSON Katja , NILSSON Jeanette , BJURSELL Gunnar
Published: July 2002

The bile salt-stimulated carboxyl ester lipase (CEL) is important for the digestion and absorption of dietary lipids, and is expressed at high levels by the exocrine pancreas and the lactating mammary gland. However, the presence of CEL in human plasma suggests that the role of CEL in lipid metabolism may stretch beyond its function in the intestinal lumen, and possibly include interactions with cholesterol and oxidized lipoproteins to modulate the progression of atherosclerosis. We have used the CEL-expressing human monocytic cell line THP-1 to investigate the transcriptional regulation of the human CEL in monocytes. Analyses of the promoter region revealed that an E-box located at −47/−52 is necessary for CEL expression. Point mutations in the E-box almost completely abolish the transcriptional activity. Electrophoretic mobility-shift assay analyses reveal that the E-box binds the upstream stimulatory factors 1 and 2, and the binding of an upstream stimulatory factor-containing complex in THP-1 cells also requires the presence of a putative nuclear receptor-binding site at −60/−66. Furthermore, we demonstrate that the E-box is also necessary for CEL expression in the pancreas and the mammary gland, although there are tissue-specific requirements for additional activating elements.
Uromodulin promoter directs high-level expression of biologically active human α1-antitrypsin into mouse urine
Authors: ZBIKOWSKA Halina M. , SOUKHAREVA Nadia , BEHNAM Reza , LUBON Henryk , HAMMOND David , SOUKHAREV Serguei
Published: July 2002

We have recently shown that the regulatory sequence of the uromodulin gene, containing the 3.7kb promoter, exon 1 and a part of exon 2, provided for kidney-specific expression of the reporter lacZ gene in transgenic mice [Zbikowska, Soukhareva, Behnam, Chang, Drews, Lubon, Hammond and Soukharev (2002) Transgenic Res., in the press]. In the present study, we generated transgenic mice harbouring the regulatory sequence of the uromodulin gene to direct the expression of human α1-antitrypsin (α1AT) into urine. Of the 13 founder mice that tested positive by PCR, seven showed the presence of the human protein in their urine. The concentration of the recombinant human (rh) α1AT in the urine, estimated by using ELISA, ranged from 0.5 to 14μg/ml in the F0-generation mice, and reached up to 65μg/ml in the F1 generation. The transgenically produced rh α1AT was found to be N-glycosylated and biologically active. The N-terminal sequence analysis confirmed the identity of the human protein and revealed that the recombinant α1AT was correctly processed with the signal peptide cleaved off. Our results demonstrate for the first time that the uromodulin regulatory sequence provides a very attractive option for the potential large-scale production of functional therapeutic proteins in livestock.
A mammalian Rho-specific guanine-nucleotide exchange factor (p164-RhoGEF) without a pleckstrin homology domain
Authors: RÜMENAPP Ulrich , FREICHEL-BLOMQUIST Andrea , WITTINGHOFER Burkhard , JAKOBS Karl H. , WIELAND Thomas
Published: July 2002

Rho GTPases, which are activated by specific guanine-nucleotide exchange factors (GEFs), play pivotal roles in several cellular functions. We identified a recently cloned human cDNA, namely KIAA0337, encoding a protein containing 1510 amino acids (p164). It contains a RhoGEF-specific Dbl homology (DH) domain but lacks their typical pleckstrin homology domain. The expression of the mRNA encoding p164 was found to be at least 4-fold higher in the heart than in other tissues. Recombinant p164 interacted with and induced GDP/GTP exchange at RhoA but not at Rac1 or Cdc42. p164-ΔC and p164-ΔN are p164 mutants that are truncated at the C- and N-termini respectively but contain the DH domain. In contrast with the full-length p164, expression of p164-ΔC and p164-ΔN strongly induced actin stress fibre formation and activated serum response factor-mediated and Rho-dependent gene transcription. Interestingly, p164-ΔN2, a mutant containing the C-terminus but having a defective DH domain, bound to p164-ΔC and suppressed the p164-ΔC-induced gene transcription. Overexpression of the full-length p164 inhibited M3 muscarinic receptor-induced gene transcription, whereas co-expression with Gβ1γ2 dimers induced transcriptional activity. It is concluded that p164-RhoGEF is a Rho-specific GEF with novel structural and regulatory properties and predominant expression in the heart. Apparently, its N- and C-termini interact with each other, thereby inhibiting its GEF activity.
Long‐term cannabis use: characteristics of users in an Australian rural area
Authors: Reilly David , Didcott Peter , Swift Wendy , Hall Wayne
Published: July 2002

Aim. To investigate the characteristics and patterns of cannabis and other drug use among long‐term cannabis users in an Australian rural area . Design. Cross‐sectional survey of a "snowball" sample of long‐term cannabis users . Setting. The North Coast of New South Wales is an area with high levels of cannabis cultivation and use, and many long‐term users . Participants. The study involved 268 long‐term cannabis users who had regularly used cannabis for at least 10 years . Measurements. A structured interview schedule obtained information on: demographics, social circumstances, patterns of cannabis and other drug use, contexts of use, perceptions about cannabis and legal involvement. Findings. The mean age of the sample was 36 years and 59% were male. The median length of regular cannabis use was 19 years. Most (94%) used two or more times a week and 60% used daily, with a median of two joints per day. Two‐thirds (67%) used cannabis in social settings and two‐thirds grew cannabis for their own use. The most common reasons for using cannabis were for relaxation or relief of tension (61%) and enjoyment or to feel good (27%). The most commonly reported negative effects were feelings of anxiety, paranoia, or depression (21%), tiredness, lack of motivation and low energy (21%) and effects of smoke on the respiratory system (18%). The majority drank alcohol (79%) and over one‐third were drinking at hazardous levels. Most were current (64%) or ex‐tobacco smokers (24%). One‐quarter (25%) had been charged with possession of cannabis, 11% for cultivation and 6% for supply, with non‐drug offences low (8% or less). Overall, three‐quarters (72%) believed that the benefits of cannabis use outweighed the risks, 21% felt there was an even balance, and 7% said cannabis had done them more harm than good. Conclusions. Among long‐term cannabis users in this Australian rural area, cannabis use was an integral part of everyday life and it was primarily used in social situations for the same reasons that alcohol use is used in the wider community.
Abrogation of the fibrotic effect of transforming growth factor‐β in dermal wound healing
Authors: Parrelli Jo M. , Meisler Natalie , Cutroneo Kenneth R.
Published: July 2002

The growth factor, transforming growth factor‐β1, which under normal circumstances promotes wound healing by stimulating local fibroblasts to produce collagen and other extracellular matrix proteins, has also been implicated as the primary causative agent of fibrosis. Because transforming growth factor‐β1 is capable of stimulating its own production by fibroblasts, its normally beneficial effects may become amplified to the point where excess extracellular matrix accumulation occurs, thereby causing abnormal scarring. Therefore, strategies that block or counter the effects of transforming growth factor‐β1 may be useful in preventing or decreasing fibrosis. One such strategy is the use of glucocorticoid steroids such as dexamethasone, which normally have the opposite effect of transforming growth factor‐β1, namely the impairment of wound healing. When used in conjunction with transforming growth factor‐β1, glucocorticoid steroids may normalize the effect of transforming growth factor‐β1 on collagen synthesis, thereby reducing excessive collagen deposition and fibrosis.
In vitro analysis of fetal fibroblast collagenolytic activity
Authors: Gould Lisa J. , Yager Dorne R. , Cohen I. Kelman , Diegelmann Robert F.
Published: July 2002

Tissue repair in the rabbit fetus is remarkably rapid and occurs without significant inflammation or excessive collagen deposition. The objective of this study was to compare the ability of rabbit fetal and adult fibroblasts to express the matrix metalloproteinases which are thought to be critical to scar tissue remodeling. In vitro, both fetal and adult rabbit fibroblasts express procollagenase messenger RNA in a constitutive manner. Mechanical disruption of fetal fibroblast monolayers caused a twofold increase in procollagenase mRNA. In contrast, the adult rabbit fibroblast procollagenase mRNA remained unchanged. The mRNA data correlated well with enzyme protein levels. Quantitation by immunoprecipitation showed a 2.3‐fold increase in fetal fibroblast procollagenase protein after mechanical injury, whereas the level in adult rabbit fibroblasts remained unchanged. However, it was noted that the constitutive levels of procollagenase mRNA and protein were higher in adult fibroblasts. Analysis of enzyme activity, by means of a fluorogenic substrate, showed that adult fibroblasts had 2.2 times more collagenase activity compared with fetal cells. After mechanical injury, the fetal fibroblast collagenase activity increased 1.3‐fold compared with 1.7‐fold in the adult fibroblasts. In contrast, fetal fibroblast gelatinase activity was 1.25 times greater than in adult cells and increased 1.4‐fold after mechanical injury, whereas the adult profile remained unchanged. Immunolocalization studies indicated that 1 hour after mechanical injury, procollagenase was produced primarily by fibroblasts along the mechanical injury ridge. By 4 hours after injury, the ridge cells began to migrate out into the open area and procollagenase was noted in adjacent cells of both adult and fetal origin. By 12 and 18 hours, all cells throughout the monolayer were expressing procollagenase. These findings show that in vitro fetal fibroblasts actually had lower levels of procollagenase, but higher levels of gelatinase compared with adult fibroblasts. The increased gelatinase expression may explain why fetal wounds do not have excessive collagen accumulation and heal without a visible scar.
Acceleration of dermal tissue repair by angiotensin II
Authors: Rodgers Kathleen , Abiko Masami , Girgis Wefki , St. Amand Karen , Campeau Joseph , Dizerega Gere
Published: July 2002

Angiotensin II is a naturally occurring peptide which has been shown to possess angiogenic properties. In the studies reported here, angiotensin II was shown to increase the proliferation of cultured bovine aortic arch endothelial cells in a concentration‐dependent manner. Acute administration of angiotensin II in Hydron accelerated the repair of dermal injuries in a full‐thickness excisional rat model. Additional studies were done to determine the best vehicle for delivery of angiotensin II to a dermal injury. Several vehicles, including 10% low‐viscosity carboxymethyl cellulose, 4% medium‐viscosity carboxymethyl cellulose, and 3% high‐viscosity carboxymethyl cellulose, were found to be effective in this regard. Daily administration of angiotensin II for days 0 to 4 after injury (day 0 being the time of surgery) was determined to provide the optimal dosage for acceleration of wound repair by angiotensin II. In addition, dose‐response studies indicated that angiotensin II accelerated wound repair in a dose‐dependent fashion with 0.03 and 0.01 µg/rat/day of angiotensin II administered on days 0 to 4 being the minimally effective and no‐effect doses, respectively. Administration of 100 µg/day of angiotensin II in 10% carboxymethyl cellulose for 5 days after injury to animals with impaired healing (steroid‐ and adriamycin‐treated rats and diabetic mice) was also found to accelerate the rate of repair. In conclusion, angiotensin II accelerated the closure of full‐thickness skin injuries in a dose‐dependent manner in normal and impaired animal models.
Single local instillation of nonviable Staphylococcus aureus or its peptidoglycan ameliorates glucocorticoid‐induced impaired wound healing
Authors: Chang Tsueng H. , Patel Mayank , Watford Alvin , Freundlich Larry , Steinberg J. J. , Levenson Stanley M.
Published: July 2002

An excess in glucocorticoid steroids, either from endogenous or exogenous sources, has been shown to inhibit wound repair. Key to this impairment is a diminution of the inflammatory response to wounding, fibroplasia, capillary formation, reparative tissue collagen accumulation, and wound breaking strength. Because a single local application at operation of nonviable Staphylococcus aureus or its peptidoglycan increases all of these processes in normal rats, we hypothesized that nonviable S. aureus and S. aureus peptidoglycan would each ameliorate glucocorticoid‐induced impaired healing. Sprague‐Dawley male rats aseptically received two 7 cm paravertebral skin incisions and underwent subcutaneous implantation of polyvinyl alcohol sponges. Two glucocorticoids were used: hydrocortisone, 8 mg intramuscularly, daily beginning 1 day before operation and continuing during the postoperative period; or a single dose of a long‐acting preparation of methylprednisolone, 6 or 8 mg intramuscularly, on the day before operation. Controls received intramuscular injections of saline solution at the same respective times. At the time of the operation, one incision and the polyvinyl alcohol sponges on one side of the animal were instilled with saline solution while the incision and sponges on the opposite side were instilled with nonviable S. aureus (hydrocortisone study) or S. aureus peptidoglycan (two methylprednisolone studies). The data showed that, at postoperative day 7, the single local application at wounding of nonviable S. aureus or S. aureus peptidoglycan increased wound breaking strength in the control rats by factors of 1.6 in the hydrocortisone experiment and 1.4 and 1.6 in the methylprednisolone studies. These treatments prevented (in hydrocortisone‐treated rats) or mitigated (in methylprednisolone‐treated rats) the glucocorticoid‐induced decrease in wound breaking strength. In addition, these treatments prevented the glucocorticoid‐induced decreases in the inflammatory (largely mononuclear cells) response to wounding and in the accumulation within the polyvinyl alcohol sponge of reparative tissue fibroblasts, capillaries, and collagen.
Cellular distribution of 5‐lipoxygenase and leukotriene receptors in postsurgical peritoneal wound repair
Authors: Chegini Nasser , Rossi Michael J. , Holmdahl Lena
Published: July 2002

Products of the eicosanoid pathways, namely prostaglandins and leukotrienes, are known to play a key role in inflammatory and immune responses, as well as other wound cellular activities of wounded tissue. The objective of this study was to determine whether surgically induced intraperitoneal and incisional wounds in the rat express 5‐lipoxygenase and contain binding sites for leukotrienes and whether their levels change during the course of healing. With the use of a specific monoclonal antibody generated against 5‐lipoxygenase, the enzyme was immunohistochemically localized in various wound cells during postsurgical days 2 to 35. The inflammatory cells within the wound were the major cell types containing 5‐lipoxygenase immunoreactive protein, followed by fibroblasts in the incisional and peritoneal fibrous adhesions, striated muscle, and the vasculature. The greatest level of immunostaining was observed during the first 2 weeks after surgery, which decreased to near unwounded levels by day 35. Light microscope autoradiographic binding studies using 3H‐leukotrienes indicated that the peritoneal/incisional wounds and unwounded tissues contain specific 3H‐leukotriene C4 and 3H‐leukotriene D4 but not 3H‐leukotriene B4 binding sites. Quantitative grain analysis (net grain density/100 µm2), representing specific 3H‐leukotriene C4 and 3H‐leukotriene D4 binding sites calculated for different cell types in the wound and unwounded regions showed that 3H‐leukotriene C4 binding was highest over the striated muscle proximal to the injury and incisional and peritoneal granulation tissue fibroblasts. The net grain density over these cells increased by 3‐, 2.5‐, and 2‐fold by day 14, respectively, and declined to the control values by day 21 after injury (p < 0.05). The pattern of 3H‐leukotriene D4 binding was similar to that observed for 3H‐leukotriene C4, but with a lower density. The grain density for 3H‐leukotriene C4 and 3H‐leukotriene D4 in arteriolar endothelial and smooth muscle cells remained unchanged. These data suggest that the products of the lipoxygenase pathway through the presence of their specific receptors may play an important role in peritoneal wound repair and adhesion formation.
Early tracheostomal healing in rabbits with use of various tracheal incisions
Authors: Whitley Rhonda , Castillo Nieva , Hassett James M. , Banyas Jeffrey , Luchette Fred A.
Published: July 2002

Various tracheal incisions (vertical, horizontal, or window) are used by surgeons for creation of a tracheostomy. The inflammatory response and healing varies with each incision and may contribute to complications such as tracheal stenosis. This study evaluates the effect of these tracheotomies on early stomal wound healing in a rabbit model. Male juvenile New Zealand rabbits underwent tracheotomy, with each animal randomized to the type of tracheal incision used (vertical, horizontal, or window). After recovery, they were killed on postoperative days 2, 4, 6, and 8, with tissue removed for histologic examination. Paraffin‐embedded stomal sections were analyzed quantitatively for amounts of granulation tissue, fibrosis, and epithelization. Groups were compared statistically using chi‐square, ANOVA, Spearman's rho, and Mann‐Whitney U tests with p less than 0.05 considered significant. Fibrosis was significantly increased in the vertical and horizontal groups when compared with the window group. This increase was statistically significant between postoperative days 2 and 4 (p < 0.05). The amount of granulation tissue was only significantly increased in the window group, whereas no difference was seen in the rate at which epithelization occurred with the various incisions. Vertical and horizontal tracheal wounds have less granulation tissue formation and more fibrosis compared with window tracheotomies during initial wound healing. This could lead to a “safer” tracheostomy tract in the early postoperative period.
Nonviable Staphylococcus aureus and its peptidoglycan stimulate macrophage recruitment, angiogenesis, fibroplasia, and collagen accumulation in wounded rats
Authors: Kilcullen Jack K. , Ly Quan P. , Chang Tsueng H. , Levenson Stanley M. , Steinberg Jacob J.
Published: July 2002

We have previously shown that local application at the time of operation of Staphylococcus aureus, nonviable S. aureus, its cell wall, or S. aureus peptidoglycan accelerates wound healing. We hypothesized that this effect is due to both direct and indirect mechanisms, among which is an increase in the inflammatory response to wounding, resulting in an increase in macrophages, angiogenesis, and fibroblasts. Twenty‐seven Sprague‐Dawley male rats were anesthetized, and two 7‐cm paravertebral skin incisions were made. Four polyvinyl alcohol sponges, two on each side, containing either 100 µl of isotonic saline or 0.5 mg of nonviable S. aureus or S. aureus peptidoglycan in 100‐µl saline were implanted subcutaneously. Nonviable S. aureus or S. aureus peptidoglycan (860 µg/cm incision) in 200‐µl saline were inoculated into the incisions at closure. The rats ate a commercial rat chow and drank tap water ad libitum throughout. After days 3 and 7 postwounding, rats were euthanized, and tissues were examined for immunohistochemical features of reparative tissue using ED‐1, Factor VIII, and vimentin antibodies, markers for monocyte/macrophages, endothelial cells, and mesenchymal cells (including fibroblasts), respectively. Incisions treated with nonviable S. aureus or S. aureus peptidoglycan showed more macrophages along and deep in the wound tract 7 days postoperatively. Nonviable S. aureus or S. aureus peptidoglycan‐treated sponges were surrounded and penetrated by much larger capsules of reparative tissue than saline‐treated sponges at both 3 and 7 days. Neutrophil influx was much greater in nonviable S. aureus or S. aureus peptidoglycan‐treated sponges, especially in central regions, and there were many more ED‐1‐stained macrophages in distinct geographic locations, specifically, the more peripheral‐cortical areas. Some clustering of macrophages occurred around areas of invasion by reparative tissue into the surrounding subcutaneous fat and within the interstices of the sponges at the interface between reparative tissue and acute inflammatory cells. In contrast, saline‐treated sponge reparative tissue had significantly fewer macrophages, much thinner and flimsy reparative tissue, with proportionately fewer macrophages clustering centrally. There were many more mesenchymal cells (notably fibroblasts) and new blood vessels and much more reparative collagen in the nonviable S. aureus or S. aureus peptidoglycan‐ treated sponges. We conclude that local application of nonviable S. aureus or S. aureus peptidoglycan at wounding induces an increased number and alteration in location of macrophages, increased influx (or proliferation) of mesenchymal cells (notably fibroblasts), and increased angiogenesis and reparative collagen accumulation, as well as increasing the overall acute inflammatory response to wounding.
Thyroid hormone regulation of Xenopus laevis metamorphosis: functions of thyroid hormone receptors and roles of extracellular matrix remodeling
Authors: Shi Yun‐Bo , Sachs Laurent M. , Jones Peter , Li Qing , Ishizuya‐Oka Atsuko
Published: July 2002

The regulatory effects of the thyroid hormone on amphibian metamorphosis is mediated by thyroid hormone receptors. Using Xenopus laevis as a model system, we and others have shown that the mRNA levels of thyroid hormone receptors and 9‐cis retinoic acid receptors, which form the functional heterodimers with thyroid hormone receptors, are regulated temporally in a tissue‐dependent manner so that high levels of their mRNAs are present in an organ when metamorphosis is occurring. By overexpressing thyroid hormone receptors, 9‐cis retinoic acid receptors, or both into developing Xenopus embryos, we have shown that both thyroid hormone receptors and 9‐cis retinoic acid receptors are required for mediating the effects of thyroid hormone on embryo development and precocious but specific regulation of the genes, which are normally regulated by thyroid hormone during metamorphosis. Analyses of the developmental expression of one class of thyroid hormone response genes, which encode extracellular matrix‐degrading metalloproteinases, suggest that extra cellular remodeling plays an important role during tissue remodeling, including cell death (apoptosis) and cell proliferation and differentiation. This effect of extracellular matrix on cell behavior has been supported directly by in vitro primary cell culture experiments, in which intestinal epithelial cells undergo thyroid hormone‐induced apoptosis, just like that during natural metamorphosis.
Effects of peripheral nerve implants on the regeneration of partially and fully innervated urodele forelimbs
Authors: Irvin Brett C. , Tassava Roy A.
Published: July 2002

This study addresses the cellular mechanism of the nerve requirement for regeneration of the urodele forelimb. Others have suggested that only the Schwann cell lineage of the blastema requires nerves for regeneration and that upon limb denervation, Schwann cells arrest in the cell cycle and produce a factor that inhibits the cycling of the remaining blastema cells. Our objective was to test this Schwann cell inhibitor model. First, pieces of peripheral nerve were implanted into partially denervated (one third of the nerve supply cut) axolotl forelimbs in an attempt to provide sufficient additional Schwann cells to increase the threshold nerve requirement above that provided by the remaining nerves. These limbs showed delayed regeneration in 68% of the cases and mild deformities, as seen by Victoria Blue staining, in 10% of the cases, as compared with control, partially denervated contralateral limbs that received grafts of muscle or frozen/thawed nerve. Second, when pieces of peripheral nerve were implanted into fully innervated newt limbs, blastema formation was limited, and regeneration was delayed in 80% of experimental cases when compared with control, contralateral newt limbs with muscle or frozen/thawed nerve implants. The results support the inhibition model and further link the need for nerves in regeneration to a possible specific requirement by Schwann cells.
Effect of electrical stimulation on foot skin perfusion in persons with or at risk for diabetic foot ulcers
Authors: Gilcreast Darlene M. , Stotts Nancy A. , Froelicher Erika S. , Baker Lucinda L. , Moss Kathryn M.
Published: July 2002

The failure of foot wounds to heal results in 54,000 people with diabetes having to undergo extremity amputations annually. Therefore, treatment is needed to speed healing in people with diabetes in order to reduce the need for amputation. This study tested the effect of high‐voltage pulsed current on foot blood flow in human beings who are at risk for diabetic foot ulcers. Neuropathy, vascular disease, Wagner Class, glucose, gender, ethnicity, and age were measured. A sample of 132 subjects was tested using a repeated‐measures design. A baseline transcutaneous oxygen level was obtained; stimulation was applied, and transcutaneous oxygen measurements were recorded at 30‐ and 60‐minute time intervals. The grouped foot transcutaneous oxygen levels decreased ({ifF = 5.66, p = .0039}) following electrical stimulation. Analysis of variance (Scheffe, p < .05) showed that initial transcutaneous oxygen was significantly higher than subsequent readings. However, oxygen response was distributed bimodally: 35 (27%) subjects showed increased transcutaneous oxygen (mean 14.8 mm Hg), and 97 (73%) experienced a decreased transcutaneous oxygen reading (mean 12.2 mm Hg). Logistic regression analysis did not explain these differences. Although this treatment appears to increase blood flow in a subset of patients, further study is needed to identify probable mechanisms for this response.
The risk of transfusion‐acquired CMV infection in seronegative solid‐organ transplant recipients receiving non‐WBC‐reduced blood components not screened for CMV antibody (1984 to 1996): experience at a single Canadian center
Authors: Preiksaitis Jutta K. , Sandhu Jatinder , Strautman Maureen
Published: July 2002

BACKGROUND: The use of CMV‐safe cellular blood components has been recommended for CMV‐sero‐ negative recipients of CMV seronegative (R–D–) solid‐ organ transplants.

STUDY DESIGN AND METHODS: The incidence of CMV infection in 281 CMV‐seronegative patients receiving renal, heart, heart–lung, lung, and liver transplants at our center between January 1984 and October 1996 was studied. The blood components that these patients received were neither WBC reduced nor screened for CMV antibody.

RESULTS: One hundred thirty‐one of 154 (85.1%) patients receiving organs from CMV‐seropositive donors (R–D+) developed CMV infection compared with three cases of presumptive transfusion‐acquired (TA) CMV infection in 127 R–D– recipients (2.4%) (relative risk, 36.0; 95% CI, 11.8‐110.4). The organ‐specific incidence of TA CMV infection in R–D– patients was as follows: renal, 0 of 57 (0%); heart, 0 of 29 (0%); heart–lung and/or lung, 1 of 6 (16.7%); and liver, 2 of 20 (10%). There was no significant difference in the transfusion requirements of CMV‐infected and ‐uninfected R–D– patients. False‐positive results were often (40%) observed when posttransfusion serum samples were used for determination of the organ donor CMV serostatus.

CONCLUSION: The low risk of TA CMV infection observed in transplant patients who received standard blood components in our study should be considered when evaluating the efficacy of programs that provide CMV‐safe blood components for this population.
HIV seroconverting donors delay their return: screening test implications
Authors: Schreiber George B. , Glynn Simone A. , Satten Glen A. , Kong Fanhui , Wright David , Busch Michael P. , Tu Yongling , Kleinman Steven H. ,   for the Retrovirus Epidemiology Donor Study
Published: July 2002

BACKGROUND: The yield of HIV p24 antigen testing implemented in March 1996 has been lower than projected. One possible explanation is that HIV seroconverting donors delay their return because of the recent practice of risk behaviors and/or signs and symptoms associated with primary infection.

STUDY DESIGN AND METHODS: From a database of 6.8‐million allogeneic donations collected at five U.S. blood centers from 1991 to 1997, 49 HIV, 21 HCV, 32 HTLV, and 44 HBsAg seroconverters with at least three donations were identified. A statistical method was developed to investigate whether the time between a donor's last negative donation and their positive donation was significantly longer than expected based on their previous return history.

RESULTS: HIV seroconverters returned on average 42 percent later than expected (p < 0.01). Although not significant, HCV seroconverters donated on average 43 percent earlier than expected. HTLV and HBsAg seroconverters did not appear to change their donation pattern around the time of seroconversion. Sixty‐three percent of the HIV seroconverters later acknowledged practicing a high‐risk behavior.

CONCLUSIONS: HIV seroconverters delay their return around the time of seroconversion and are thus less likely to be recently infected. Unique among HIV seroconverters, this observation provides a possible explanation for the lower than expected yield of HIV p24 antigen testing and suggests that NAT may have a similar low yield.