bluuebunny/crossref_metadata_2025 huggingface dataset:
Effect of recombinant human deoxyribonuclease on the expression of cell adhesion molecules of thawed and processed cord blood hematopoietic progenitors
Authors: Beck C , Nguyen XD , Klüter H , Eichler H
Published: March 2003

Abstract: Background and objectives: The integrity of granulocytic cells and platelets is compromised within cryopreserved stem cell transplants, and consequent DNA release during the thawing procedure can therefore lead to clotting phenomena or microaggregate formation and that in turn may cause loss of progenitor cells. To circumvent this problem a new processing protocol was introduced using recombinant human deoxyribonuclease (rhDNase) to prevent cell aggregate formation. In addition, the impact of this new processing protocol on CD34+ umbilical cord blood (UCB) cells was assessed.

Materials and methods: Fifty samples derived from 7 buffy coat (BC) volume reduced UCB units were cryopreserved, thawed, and processed with washing solutions that were supplemented with rhDNase in various concentrations. Thereafter, clotting and microaggregate formation was scored microscopically. In addition, expression of the adhesion molecules leukocyte function‐associated antigen 1 (LFA‐1, n = 6), intercellular adhesion molecule‐1 (ICAM‐1, n = 11), and L‐selectin (n = 11) on CD34+ UCB cells was analyzed by flow cytometry after incubating the samples with either dimethyl sulfoxide (DMSO) 5.5%, rhDNase 10 or 50 U/mL, or a combination of DMSO 5.5% and rhDNase 50 U/mL.

Results: At a minimal concentration of 10 U rhDNase/mL, clotting or microaggregate formation could be prevented for all tested samples, whereas cell clots could be observed for concentrations up to 8 U/mL. The expression of adhesion molecules on untreated CD34+ UCB cells (L‐selectin: 64.6 ± 18.8%; LFA‐1: 62.6 ± 7.5%; ICAM‐1: 14.8 ± 4.1%) did not show any significant difference compared with cells that were incubated with up to 50 U/mL rhDNase (L‐selectin: 62.2 ± 19.3%; LFA‐1: 63.1 ± 5.9%; ICAM‐1: 17.5 ± 6.7%). However, after a combined treatment with DMSO 5.5% and rhDNase 50 U/mL, a slight but significant decrease in L‐selectin expression could be observed (P < 0.03).

Conclusion: The supplementation of rhDNase to a final concentration of 10 U/mL cell suspension proved to be effective in preventing clot formation under the conditions examined and did not lead to decreased expression levels of adhesion molecules. We therefore recommend the use of rhDNase for the prevention of clot formation and cell loss during the processing of thawed UCB transplants.
Expression of stratum corneum chymotryptic enzyme in relation to other markers of epidermal differentiation in a skin explant model
Authors: Ekholm E. , Egelrud T.
Published: March 2003

Abstract: The serine proteinase stratum corneum chymotryptic enzyme (SCCE) has been proposed to be involved in the degradation of intercellular cohesive structures in cornified squamous epithelia in the process of desquamation. Since SCCE is expressed late in epidermal differentiation and is found at all sites where there is a formation of cornified epithelia it also serves as a marker for terminal epidermal differentiation. Earlier studies have shown that the link between expression and the formation of cornified cells may be stronger for SCCE than for other well characterized markers of epidermal differentiation. In an attempt to further elucidate the regulation of SCCE expression we have in this study compared the expression of SCCE with the expression of keratin 10, filaggrin and involucrin in an in vitro model with skin explants cultured for various periods of time on de‐epidermized dermis at the liquid–air interface. The markers were analysed by means of immunohistochemistry. We found that the expression of SCCE preceded the expression of keratin 10 and filaggrin. In contrast to involucrin, which was expressed by all suprabasal keratinocytes, SCCE was expressed only by high suprabasal cells. Our results indicate that the expression of SCCE may be regulated in a way that differs from the regulation of the expression of keratin 10, filaggrin and involucrin.
Novel keratin 16 mutations and protein expression studies in pachyonychia congenita type 1 and focal palmoplantar keratoderma
Authors: Smith F. J. D. , Fisher M. P. , Healy E. , Rees J. L. , Bonifas J. M. , Epstein Jr E. H. , Tan E. M. L. , Uitto J. , McLean W. H. I.
Published: March 2003

Abstract: Pachyonychia congenita type 1 (PC‐1) is an autosomal dominant ectodermal dysplasia characterized by nail dystrophy, focal non‐epidermolytic palmoplantar keratoderma (FNEPPK) and oral lesions. We have previously shown that mutations in keratin 16 (K16) cause fragility of specific epithelia resulting in phenotypes of PC‐1 or FNEPPK alone. Here, we report 2 novel mutations in K16 causing distinct phenotypes. A heterozygous missense mutation (L124R) was detected in a kindred with PC‐1. In a family where mild FNEPPK was the only phenotype, a 23 bp deletion and a separate 1 bp deletion downstream were found in exon 6: [1244–1266del; 1270delG]. At the protein level, these mutations remove 8 residues and substitute 2 residues in the helix termination motif (HTM) of the K16 polypeptide. The HTM sequence is conserved in all known intermediate filament proteins and for convenience, this complex mutation was designated ΔHTM. Transient expression of K16 cDNAs carrying either the L124R or the ΔHTM mutation in epithelial cell line PtK2 produced aggregation of the keratin cytoskeleton. However, the aggregates observed with the ΔHTM mutation were morphologically different and appeared to be less disruptive to the endogenous cytoskeleton. Therefore, loss of the HTM sequence may render this mutant K16 less capable of contributing to filament assembly and decrease its dominant‐negative effect, resulting in the milder FNEPPK phenotype.
Modulation of endothelin‐1 in normal human keratinocytes by UVA1/B radiations, prostaglandin E2 and peptidase inhibitors
Authors: Pernet I. , Mayoux C. , Trompezinski S. , Schmitt D. , Viac J.
Published: March 2003

Abstract: In the skin, keratinocytes synthesize and secrete endothelin‐1 (ET‐1), a potent vasoconstrictor peptide which acts also as a growth factor for most skin cells. The aim of the study was to test the effects of UVA1 and the associations UVA1/B on the expression of ET‐1 in normal human keratinocytes and to determine whether exogenously added prostaglandin E2 (PGE2) regulated ET‐1 expression. As ET‐1 is susceptible to degradation, we also evaluated whether ET‐1 secretion was modulated by peptidase inhibitors. Our results showed that UVA1 (365 nm) did not modify the levels of preproET‐1 mRNA and protein. Moreover, the associations UVA1+UVB or UVB+UVA1 down‐regulated the overexpression of secreted ET‐1 induced by UVB alone. PGE2 at 10−5 M reduced the expression of ET‐1 at the mRNA and protein levels but did not exert any significant modification at lower concentrations from 10−10 to 10−6 M. Phosphoramidon, an endothelin converting enzyme (ECE) inhibitor, drastically decreased the amount of ET‐1 accumulating in the culture medium in basal conditions or after UVB irradiation. Conversely, thiorphan, a specific inhibitor of neutral endopeptidase (NEP), rather increased the levels of ET‐1 secretion mainly after UVB irradiation. Taken together, the results showed that normal human keratinocytes secrete and partly degrade ET‐1 through ECE and NEP pathways and pointed out a differential regulation of ET‐1 by UVB and UVA1 radiations without any noticeable role for PGE2.
The nude gene and the skin
Authors: Schlake T.
Published: March 2003

Abstract: The nude mutation has been known for a long time. Nevertheless, the gene responsible for the defect has been identified only recently. It encodes a transcriptional activator of the family of forkhead proteins mainly expressed in thymic epithelium and distinct keratinocyte populations in the epidermis and hair follicles. The present review focuses on the molecular and functional characterization of the nude gene and its product and gives an overview as to its role in skin biology and the first identified target genes in the skin. In addition, evolutionary aspects are highlighted stressing the importance of such investigations for a comprehensive understanding of the nude gene product and the regulation of its expression. Furthermore, these studies give a hint as to when the nude gene has occurred first and how it has developed in molecular and functional terms since then.
Topical synthetic inhibitor of matrix metalloproteinases delays epidermal regeneration of human wounds*
Authors: Ågren M. S. , Mirastschijski U. , Karlsmark T. , Saarialho‐Kere U. K.
Published: March 2003

Abstract: Matrix metalloproteinases (MMPs) degrade extracellular proteins during epithelialization of wounds. To evaluate the biological significance of MMPs in epidermal healing, the synthetic broad‐spectrum MMP inhibitor GM 6001 (also called Galardin and Ilomastat) was applied topically to standardized human wounds. GM 6001 (10 μg/μl) or vehicle alone was applied every second day onto 4 de‐roofed 6 mm suction blister wounds on the volar forearm of healthy male volunteers for 12 days. GM 6001 delayed healing by 2–4 days as assessed macroscopically and microscopically. In situ hybridization or immunohistochemistry showed that MMP‐1 (interstitial collagenase) was present in and MMP‐2 (gelatinase A) close to laterally migrating keratinocytes whereas MMP‐9 (gelatinase B) was seen during maturation of new epidermis. MMP‐1 was undetectable in blister roofs (normal epidermis) and found in low levels in normal skin. Total MMP‐1 activities increased about 100‐fold in wounds, independent of treatment, compared to normal skin as analyzed by specific ELISA‐based activity assay. By gelatin zymography, MMP‐2, but not MMP‐9, was detected in blister roofs and wound healing was associated with increased active MMP‐2 and latent MMP‐9 levels. GM 6001 prevented activation of MMP‐2 and increased latent MMP‐9 levels. GM 6001 delayed re‐appearance of laminin‐5, the synthesis of which correlated with epidermal regeneration. Restoration of stratum corneum, measured indirectly by transepidermal water loss, was also impaired (P<0.05) in the GM 6001 group. In conclusion, pharmacological MMP inhibition delayed epidermal regeneration in vivo, suggesting that MMPs are required to restore epidermis after epidermal ablation in humans.
Characterization of vibrissa germinative cells: transition of cell types
Authors: Osada A. , Kobayashi K.
Published: March 2003

Abstract: Germinative cells, small cell masses attached to the stalks of dermal papillae that are able to differentiate into the hair shaft and inner root sheath, form follicular bulb‐like structures when co‐cultured with dermal papilla cells. We studied the growth characteristics of germinative cells to determine the cell types in the vibrissa germinative tissue. Germinative tissues, attaching to dermal papillae, were cultured on 3T3 feeder layers. The cultured keratinocytes were harvested and transferred, equally and for two passages, onto lined dermal papilla cells (LDPC) and/or 3T3 feeder layers. The resulting germinative cells were classified into three types in the present experimental condition. Type 1 cells grow very well on either feeder layer, whereas Type 3 cells scarcely grow on either feeder layer. Type 2 cells are very conspicuous and are reversible. They grow well on 3T3 but growth is suppressed on LDPC feeder layers. The Type 2 cells that grow well on 3T3 feeder layers, however, are suppressed when transferred onto LDPC and the Type 2 cells that are suppressed on LDPC begin to grow again on 3T3. The transition of one cell type to another in vitro and the cell types that these germinative cell types correspond to in vivo is discussed. It was concluded that stem cells or their close progenitors reside in the germinative tissues of the vibrissa bulb except at late anagen–early catagen.
Intermediate or chronic cutaneous leishmaniasis: leukocyte immunophenotypes and cytokine characterisation of the lesion
Authors: Diaz N. L. , Zerpa O. , Ponce L. V. , Convit J. , Rondon A. J. , Tapia F. J.
Published: March 2003

Abstract: The American cutaneous forms of leishmaniasis include immune‐responder individuals with localised cutaneous leishmaniasis (LCL) and non‐responder individuals with diffuse cutaneous leishmaniasis (DCL). Patients with intermediate or chronic cutaneous leishmaniasis (ICL) have increased morbidity due to the length of their illness, atypical forms and areas of compromise. In the present study, we evaluated the expression of the leukocyte antigens (CD4, CD8, CLA: cutaneous lymphocyte antigen, CD69, CD83 and CD1a) and cytokines (IFN‐γ, IL‐4, IL‐10 and TGF‐β1) in the lesions of patients with ICL (n = 18) using an immunocytochemical procedure. ICL results were compared with the information for LCL (n = 19) and DCL (n = 4). The numbers of CD4+ and CD8+ T cells in ICL were similar to those of LCL lesions, but significantly different (P ≤ 0.05) from DCL lesions. LCL lesions have about half the numbers of early activated CD69+ cells as ICL, but most are CLA+ skin homing memory T cells, whereas ICL lesions have the highest number of CD69+ T cells, but about one‐third of these cells expressed CLA. This suggests that the granuloma of ICL patients contains many activated T cells that are unprimed to cutaneous‐launched antigens, thus contributing to an aberrant immune response. In contrast, DCL granulomas presented the lowest numbers of activated CD69+ and CLA+ cells, associated with the characteristic tolerogenic state of these patients. The immunolocalisation of cytokines showed a mixed cytokine pattern in ICL lesions with many positive cells for IL‐10, TGF‐β1, IL‐4 and IFN‐γ, with a preponderance of the first two, and different from the prevalent Th1 and Th2 responses associated with LCL and DCL lesions, respectively. CD1a+ Langerhans cells were decreased (P ≤ 0.05) in both ICL (271 ± 15 cells/mm2) and DCL (245 ± 19 cells/mm2) as compared to LCL (527 ± 54 cells/mm2) epidermis. The percentage of IL‐10+ epidermal Langerhans cells in ICL (33.69), from the total CD1a+ population, was higher than in LCL (17.45). In addition, fewer CD83+ primed Langerhans cells were present in ICL epidermis. The diminished participation of epidermal Langerhans cells, causing a defective signalling by the epidermis, in ICL lesions may account for the tissue‐damaging state observed in these patients.
Type‐1 steroid 5α‐reductase is functionally active in the hair follicle as evidenced by new selective inhibitors of either type‐1 or type‐2 human steroid 5α‐reductase
Authors: Gerst C. , Dalko M. , Pichaud P. , Galey J. B. , Buan B. , Bernard B. A.
Published: March 2003

Abstract: Steroid 5α‐reductase catalyzes the reduction of testosterone (T) into the very potent androgen dihydrotestosterone (DHT). The different tissue expression patterns of the two isoforms of 5α‐reductase, namely type‐1 and type‐2 5α‐reductase (5α‐R1 and 5α‐R2, respectively), have prompted studies directed towards the synthesis of selective 5α‐R1 or 5α‐R2 inhibitors. In this present work, we have performed a structure/activity study on the inhibitory potential of indole carboxylic acids against hair follicle 5α‐reductase activity. We have demonstrated that this class of molecules were potent inhibitors of either 5α‐R1 or 5α‐R2 or both depending on (i) substituents in positions 4, 5 or 6 and (ii) the presence of a free carboxylic group. We have also found that only 5α‐R1 or 5α‐R1/R2 inhibitors were able to inhibit 5α‐reductase activity in plucked hairs from female volunteers or in freshly isolated female hair follicles, selective 5α‐R2 inhibitors being inactive.
Induction of HSP27 nuclear immunoreactivity during stress is modulated by vitamin C
Authors: Boxman Ingeborg L. A. , Kempenaar Johanna , De Haas Ellis , Ponec Maria
Published: March 2003

Abstract: For the investigation of the skin irritancy potential of chemicals in an in vitro model it is necessary to have sensitive endpoints that predict the effects of those compounds on native human skin. Recently, we have identified that 27‐kDa heat shock protein (HSP27) can serve as a sensitive marker of skin irritation, as exposure of human skin to sodium lauryl sulfate (SLS) both in vitro and in vivo induced relocalization of HSP27 from the cytoplasm to the cell nucleus. The aim of the present study was to determine whether nuclear localization of HSP27 could be used as a parameter for evaluation of potential skin irritants in screening assays in vitro. For this purpose, human skin equivalent consisting of epidermis reconstructed on de‐epidermized dermis was exposed to SLS or UV light. Stress‐induced nuclear relocalization of HSP27 was observed in excised skin exposed to SLS or UV light and in reconstructed epidermis only when the latter was generated in the absence of vitamin C. The omission of vitamin C results in an impaired barrier function. In the presence of vitamin C, however, the barrier function was comparable with excised skin, suggesting that vitamin C may control the response to stress in the reconstructed epidermis. Besides the presence of vitamin C, the response of skin equivalents may strongly depend on other conditions under which they are generated, because the stress‐induced HSP27 relocalization was not detected in the commercially available epidermal kit EpiDerm. The results of the present study show that HSP27 nuclear staining can serve as a sensitive marker for skin irritation or cellular stress in excised skin as well as in certain well‐characterized human skin equivalents in vitro.
Estuarine predation on radiotagged wild and domesticated sea trout (Salmo trutta L.) smolts
Authors: Dieperink C. , Pedersen S. , Pedersen M. I.
Published: March 2003

Abstract – Avian predation on emigrating wild and domesticated sea trout smolts was investigated in a fjord in the western Baltic Sea. In April 1997, 50 domesticated and 50 wild smolts were intraperitoneally tagged with radio‐transmitters and released in a small coastal stream. Predation was recorded by signal interception in an estuarine breeding colony of cormorants and herons near the outlet of the stream. Of the 78 emigrating smolts, 51 (65%) were recorded as eaten. Predation rates were significantly higher among small than large smolts and significantly higher among domesticated smolts. The first 2 days after entering the sea, both wild and domesticated smolts suffered a severe daily predation rate (range 20–34%). The results support the hypothesis of a transient period immediately after exposure to full‐strength sea water, where smolts experience an elevated risk of predation. A transient increase in postsmolt mortality may be found also in moderately saline environments (20–23 ppt).
Denil fishway utilization patterns and passage of several warmwater species relative to seasonal, thermal and hydraulic dynamics
Authors: Bunt C. M. , Van Poorten B. T. , Wong L.
Published: March 2003

Abstract – Two different Denil fishways on the Grand River, Ontario, were used as check‐points to evaluate the upstream movement of fishes past a low‐head weir and to examine the proportions and inferred swimming performance of non‐salmonid warmwater fishes that used each fishway type. Traps installed at fishway exits were used to collect fish during 24‐hour sampling periods, over 40–51 days each year, from 1995 to 1997. Passage rates, size selectivity, water temperature, water velocity and turbidity for the periods of maximum passage for each year were examined. General species composition from trap samples shifted from catostomids to cyprinids to ictalurids to percids and centrarchids, with some overlap, as water temperatures increased from 8 °C to 25 °C in the spring and early summer. Water depths, and therefore water velocities in each fishway, were independent of river discharge due to variable accumulations of debris on upstream trash‐racks. Relationships between the water velocity and the swimming and position‐holding abilities of several species emerged. Turbidity was directly related to river discharge and precipitation events, and many species demonstrated maximum fishway use during periods of increased turbidity. This study 1) provided evidence of strongly directional upstream movements among several species that were previously considered non‐migratory and 2) describes physical and hydraulic conditions during fishway use for 29 non‐salmonid fish species.
Chinook salmon impede Atlantic salmon conservation in Lake Ontario
Authors: Scott Robert J. , Noakes David L.G. , Beamish F. W. H. , Carl Leon M.
Published: March 2003

Abstract– Non‐native species can have substantial impacts on successful restoration of native species. Here, we examined effects of chinook salmon (Oncorhychus tshawytscha), an exotic species introduced to Lake Ontario to enhance recreational angling, on reintroduced Atlantic salmon (Salmo salar) in a Lake Ontario tributary stream. Field enclosure studies revealed that adult Atlantic salmon activity rate was elevated, nest establishment delayed and mortality rates higher in the presence of chinook salmon. These results suggest that chinook salmon in Lake Ontario streams during fall spawning could impede successful re‐establishment of Atlantic salmon in the lake.
Anaerobic neutrophil‐dependent killing of Actinobacillus actinomycetemcomitans in relation to the bacterial leukotoxicity
Authors: Johansson Anders , Sandström Gunnar , Claesson Rolf , Hänström Lennart , Kalfas Sotos
Published: March 2003

The effect of leukotoxin on the interaction of Actinobacillus actinomycetemcomitans with human polymorphonuclear leukocytes (PMNL) was studied under anaerobic conditions with strains able to produce high or low amounts of leukotoxin. PMNL morphology, phagocytosis and killing were examined by transmission electron microscopy and bioassays, respectively. At ratios of ≥25 bacteria/PMNL, a highly toxic A. actinomycetemcomitans strain completely destroyed the PMNL within 7 min, resulting in bacterial survival. Lowering the bacteria/PMNL‐ratio enabled phagocytosis and killing of this highly toxic strain. A. actinomycetemcomitans strains with low leukotoxicity were effectively killed by PMNL under any condition. Presence of specific antibodies against A. actinomycetemcomitans or of anti‐leukotoxin serum protected PMNL from being injured and allowed phagocytosis to occur. Pre‐incubation of the leukotoxic strain with Porphyromonas gingivalis, a bacterium that destroys leukotoxin, abolished lysis of PMNL and inhibited phagocytosis of A. actinomycetemcomitans. The results show that leukotoxin may protect A. actinomycetemcomitans against phagocytosis by human PMNL. The protection occurs at a population level and in relation to the bacterial load. The size of the bacterial population required to counteract phagocytosis is dependent on the leukotoxin production of the strain.
Effect of short chain fatty acids on human gingival epithelial cell keratins in vitro
Authors: Pöllänen Marja T. , Salonen Jukka I.
Published: March 2003

Hemidesmosomal attachment of the junctional epithelial cells to the tooth and the ability of the attached cells to divide are essential features of the healthy dentogingival junction. Short chain fatty acids are bacterial metabolites associated with gingival inflammation and periodontal pockets. In vitro, short chain fatty acids have been shown to inhibit epithelial cell division and increase the density of their keratin filaments. This study examined these keratin changes by making use of human gingival keratinocyte cultures, gel electrophoresis and Western blot. Short chain fatty acids, butyrate and propionate, increased the relative amount of keratin proteins in the cells, most strikingly keratin K17. The distribution of K17 was further studied in a culture model for human junctional epithelium and in gingival biopsies. In butyrate‐treated cultures of junctional epithelium, K17 expression was markedly increased and extended to the basal cells and to the cells mediating the attachment of the explant to the substratum. In clinically healthy gingiva, K17 was expressed predominantly in sulcular epithelium. The dividing basal cells and the cells attached to the tooth were negative. In advanced periodontitis, a strong reaction for K17 was localised to the pocket epithelium. The inhibition of epithelial cell division and the simultaneous upregulation of K17 in vitro, and the strong expression of this protein in detached pocket epithelium suggest a role for the short chain fatty acids in the degenerative process that leads to subgingival advancement of pathogens and, eventually, to periodontal pocket formation.
Antimicrobial resistance with focus on oral beta‐lactamases
Authors: Handal Trude , Olsen Ingar
Published: March 2003

Over the last 10–15 yr antibiotic resistance has increased in the oral microflora. The beta‐lactam antibiotics, i.e., penicillins and cephalosporins, are the most frequently used antimicrobial agents. Unfortunately, the efficiency of these drugs is increasingly being challenged by the emergence of resistant bacteria, which is mainly due to their production of beta‐lactamases. In this paper, mechanisms of antibiotic resistance are reviewed, with emphasis on beta‐lactamases. This review also discusses how the presence of beta‐lactamases in oral microorganisms may affect the treatment of oral diseases. Dentists can influence the emerging global crisis of antibiotic resistance by carefully evaluating the indications for antibiotic treatment. General guidelines for when and how to use antibiotics in dentistry are reviewed.
Etiologic factors influencing the prevalence of demarcated opacities in permanent first molars in a group of Swedish children
Authors: Jälevik Birgitta , Norén Jörgen G. , Klingberg Gunilla , Barregård Lars
Published: March 2003

The aim of this study was to determine possible etiological factors for a developmental enamel defect, i.e. demarcated opacities, affecting the permanent first molars. A questionnaire about possible etiological factors of enamel developmental defects was filled in by the parents of 8‐yr‐old children (n=516) prior to a dental examination of permanent teeth. Demarcated opacities of permanent first molars had been found in 18.4% of the children in a previous study of these children. Fifteen % had more than one tooth affected indicating systemic causation. Questions were asked about mother's health and medication during pregnancy, birth complications, health and medication of the child during the first 3 yr of life, breast‐feeding, heredity, and fluoride supplements. The affected children, especially the boys, were reported to have had more health problems, in particular asthma (but only 4 cases), during the first year of life. Use of antibiotics was also more common among the affected children, but owing to a strong co‐variance with health problems these factors could not be separated. Breast‐feeding history was similar in children with and without enamel defects. The etiology of hypomineralized first molars is not yet fully understood, but based on the results of this retrospective study, health problems in infancy, especially respiratory diseases, seem to be involved.
Co‐adhesion and removal of adhering bacteria from salivary pellicles by three different modes of brushing
Authors: Yang J. , Bos R. , Belder G. F. , Busscher H. J.
Published: March 2003

This study compares removal of pairs of co‐adhering and non‐co‐adhering oral actinomyces and streptococci by hand, electric and sonic brushing from salivary pellicles. In addition, re‐deposition of a co‐adhering and non‐co‐adhering streptococcal strain to brushed pellicles was studied. First, actinomyces were allowed to adhere to a pellicle in a flow chamber, after which streptococci suspended in saliva were perfused through the chamber at 33°C. Pellicles with adhering bacterial pairs were brushed and the number of bacteria remaining determined. Whereas sonic brushing removed nearly all adhering bacteria, greater numbers of larger aggregates of the co‐adhering pair, involving Streptococcus oralis J22, were left behind after hand and electric brushing than of the non‐co‐adhering pair with Streptococcus sanguis PK1889. Re‐deposition of streptococci to electrically and sonically brushed pellicles, however, was two‐fold higher for the co‐adhering pair than for the non‐co‐adhering pair. This demonstrates a role for co‐adhesion in de novo plaque formation. Removal by the three different modes of brushing was not affected by the presence of fluoride, indicating that fluoride is not able to disrupt calcium bridges between co‐adhering pairs. In contrast, in the presence of lactose only small aggregates of co‐adhering pair were left behind.
Factor structure of the Dental Beliefs Survey in a dental phobic population
Authors: Kulich Károly R. , Berggren Ulf , Hakeberg Magnus , Gustafsson Jan‐Eric
Published: March 2003

The aim of this study was to investigate the factor structure of the 15‐item Dental Beliefs Survey (DBS) in a population of dental phobic patients (n=362). Exploratory (EFA) and confirmatory (CFA) factor analyses were used. The EFA indicated three factors: ‘communication’, ‘trust’, and ‘fear of negative information’. However, the EFA further suggested a general factor with all 15 items. By using the CFA, five factors were found based on the EFA solution and the original DBS dimensions. A general factor, ‘social interaction distress in dental treatment’, and four more narrow dimensions, ‘communication’, ‘trust’, ‘fear of negative information’ (originally labelled ‘belittlement’), and ‘lack of control’, was the most adequate result with regard to theoretical and statistical properties. However, some items in the factors were partly different from the original version of the DBS. In conclusion, the DBS attempts to measure a complex phenomenon with regard to patients' perceptions and attitudes to dental care. This study has revealed psychometric properties of the DBS in a population of dental phobic patients. The important finding was a general dimension, which suggests the use of DBS as an overall measure of dental beliefs. However, more research is needed in epidemiological and clinical studies with non‐phobic individuals.
Influence of methodological parameters on human jaw‐stretch reflexes
Authors: Wang Kelun , Svensson Peter
Published: March 2003

The influence of methodological parameters and experimental conditions on the human jaw‐stretch reflex was studied in healthy subjects in order to develop a reliable tool for investigation of the excitability of motoneuron pool. Short‐latency excitatory reflex responses were evoked by a custom‐made stretch device with the subjects biting on a jaw‐bar with their front teeth. The displacement and ramp time of the stretches were accurately controlled and automatically triggered by a computer. The reflex responses were measured in the surface electromyogram (EMG) of the masseter and anterior temporalis muscles with online monitoring of the clenching level. The peak‐to‐peak amplitude of the jaw‐stretch reflex was shown to be proportional to the level of EMG activity during isometric contractions, to increase proportionally with increasing stretch displacement at a given ramp time, and to decrease proportionally with increasing ramp time at a given stretch displacement. There were no significant differences in the reflex amplitude between repeated recordings within one session or between different sessions. Local anesthetic around the lower incisors as well as the upper incisors had no significant influence on the reflex amplitude. However, different biting positions on the bars of the stretch device significantly influenced the amplitude of the stretch reflex.
Application of quantitative light‐induced fluorescence to monitor incipient lesions in caries‐active children. A comparative study of remineralisation by fluoride varnish and professional cleaning
Authors: Tranæus Sofia , Al‐Khateeb Susan , Björkman Stein , Twetman Svante , Angmar‐Månsson Birgit
Published: March 2003

The aim of this study was to apply the quantitative light‐induced fluorescence (QLF) method in a randomised controlled study, comparing treatment with fluoride varnish and professional tooth cleaning for remineralisation of white spot lesions in caries‐active adolescents. In the fluoride varnish group (n=13; 32 lesions), professional tooth‐cleaning was followed by application of fluoride varnish at the beginning of the study, after 1 wk, and then once every 6 wk for 6 months. The other group (n=18; 30 lesions) underwent professional tooth‐cleaning once every 6 wk for 6 months. Enamel fluorescence was measured at baseline and at each visit. In the fluoride varnish group there was a significant change over time (baseline: 6 months) for both lesion area: and average change in fluorescence (decreased lesion area and increased fluorescence radiance). The corresponding changes in the professional tooth‐cleaning group were not significant. There was a significant difference in average change in fluorescence between the two test groups. For lesion area, there was no significant difference, but a tendency towards a difference between the test groups. It was concluded that (a) the QLF method is a sensitive method, suitable for longitudinal quantification of incipient caries lesions on smooth surfaces; and (b) that repeated fluoride applications had a favourable effect on the remineralisation of white spot lesions as measured after 6 months.
A 3‐year comparison of dental anxiety treatment outcomes: hypnosis, group therapy and individual desensitization vs. no specialist treatment
Authors: Moore Rod , Brødsgaard Inger , Abrahamsen Randi
Published: March 2003

Outcomes of hypnotherapy (HT), group therapy (GT) and individual systematic desensitization (SD) on extreme dental anxiety in adults aged 19–65 yr were compared by regular attendance behaviors, changes in dental anxiety and changes in beliefs about dentists and treatment after 3 yr. Treatment groups were comparable with a static reference control group of 65 anxious patients (Dental Anxiety Scale ≥ 15) who were followed for a mean of nearly 6 yr. After 3 yr, 54.5% of HT patients, 69.6% of GT patients and 65.5% of SD patients were maintaining regular dental care habits. This was better than the 46.1% of the reference group, who reported going regularly to the dentist again within the cohort follow‐up period, and 38.9% of a control subgroup with observation for 3 yr. Women were better regular attenders than men at 3 yr. Specialist‐treated regular attenders were significantly less anxious and had more positive beliefs than regular attenders from reference groups. There were few differences between HT, GT and SD after 3 yr. It was concluded that many patients can, on their own, successfully start and maintain regular dental treatment habits with dentists despite years of avoidance associated with phobic or extreme anxiety. However, it also appears that these patients had less success in reducing dental anxiety and improving beliefs about dentists long‐term than did patients who were treated at the specialist clinic with psychological strategies.
Immunohistochemical study on pulpal response in rat molars after cavity preparation by Er:YAG laser
Authors: Tanabe Keita , Yoshiba Kunihiko , Yoshiba Nagako , Iwaku Masaaki , Ozawa Hidehiro
Published: March 2003

While Er:YAG laser systems are in extensive use for caries removal and cavity preparation, the effects of such treatment on pulp tissue remain unclear. This study evaluates these systems using immunohistochemical methods and compares the results with information gained from treatment using conventional burs. Cervical cavities were prepared in the upper first molars of rats, using either an Er:YAG laser or a conventional tungsten‐carbide bur. At intervals of 5 min, 6 h, 12 h, 1 d, 3 d and 7 d after cavity preparation, the teeth were processed for immunohistochemical analyses of tissue non‐specific alkaline phosphatase, OX6‐positive major histocompatibility complex class II antigen‐expressing cells and PGP 9.5‐immunoreactive nerve fibers. DNA fragmentation was detected by the terminal deoxynucleotidyl transferase‐mediated dUTP nick end labeling (TUNEL) method. Tissue non‐specific alkaline phosphatase was observed mainly in the subodontoblastic layer under the cavity lesion, from 5 min, in both groups. The immunoreactivity was more pronounced in the laser group, but by 7 d no significant differences were recognizable. At 12 h, TUNEL‐positive cells were detected around the odontoblastic layer in both groups. From 3 d to 7 d, a limited number of positive cells were still visible in the group that underwent standard treatment. Clear similarities in the distribution patterns of OX6‐immunopositive cells and PGP 9.5‐immunoreactive nerve fibers were also noted. From 12 h to 1 d, OX6‐positive cells accumulated along the pulp–dentin border, extending their processes into the dentinal tubules. Numerous bead‐like PGP 9.5‐immunoreactive nerve fibers were observed under the odontoblastic layer at 7 d. These results demonstrated that there was no appreciable difference in the manner in which pulp tissue responded to treatment with either Er:YAG laser or a conventional drill. This would seem to indicate the usefulness of the Er:YAG laser system in the removal of caries and cavity preparation.
Influence of dentifrices and dietary components in saliva on wettability of pellicle‐coated enamel in vitro and in vivo
Authors: Van Der Mei H. C. , White D. J. , Kamminga‐Rasker H. J. , Knight J. , Baig A. A. , Smit J. , Busscher H. J.
Published: March 2003

In vitro salivary pellicles were found to be less hydrophobic by water contact angles than clinically formed pellicles. In this study, water contact angles were measured on enamel coated with pellicles adsorbed from reconstituted human whole saliva (RHWS) and after exposure to dentifrices or dietary components. In addition, adhesion of Streptococcus oralis J22 to pellicles formed from RHWS with minor amounts of milk added and after exposure to dentifrices was studied. Exposure of RHWS‐pellicles to milk or salad oil yielded an increase in the hydrophobicity of in vitro pellicles to values observed in vivo, but a decrease was seen after exposure to a sugar solution. Pellicles formed from saliva with 0.4% milk added attracted less S. oralis cells than pellicles formed in the absence of milk components. Exposure of pellicles formed from saliva with milk added to various dentifrices had a variable effect on bacterial adhesion: markedly lower numbers of adhering S. oralis were found for a dentifrice with NaF, but exposure to dentifrices containing SnF2 or hexametaphosphate showed slightly increased adhesion. In summary, dietary components have influence on the hydrophobicity of enamel pellicles, while combinations of dietary components and dentifrices certainly influence the adhesiveness of the pellicles. The effects of dietary components on pellicle conditioning film should be taken into consideration in research on the development of ingredients to control intraoral surface chemistry and microbiology, as well as in the development of oral products.
The German version of the Oral Health Impact Profile – translation and psychometric properties
Authors: John Mike T. , Patrick Donald L. , Slade Gary D.
Published: March 2003

The need for cross‐culturally adapted oral‐health specific health outcome measures is increasingly recognized in Germany. Following accepted cultural adaptation technique guidelines, we report the development of the German version of the Oral Health Impact Profile (OHIP). The original 49 items were translated using a forward–backward method. A de novo development of German items established content validity. A priori hypothesized associations between the OHIP summary score and self‐reported oral health and five oral disorders were confirmed in a random sample of the general population (n = 163, age 20–60 yr). These associations were interpreted as support for construct validity. The instrument's responsiveness, as indicated by a mean OHIP summary score change from 45.0 to 28.3, was established in 67 consecutive patients treated for temporomandibular disorder pain (age 19–85 yr; 72% women). Test‐retest reliability was demonstrated by intraclass correlation coefficients of 0.63–0.92 for dimensions and summary scores (convenience sample, n = 30, age 18–85 yr; 53% women). Internal consistency was high (Cronbach's α > 0.74). Sufficient discriminative and evaluative psychometric properties of the Oral Health Impact Profile German version (OHIP‐G) make the instrument suitable for assessment of oral health‐related quality of life in cross‐sectional as well as longitudinal studies.
Deficiency of Decidual IL‐10 in First Trimester Missed Abortion: A Lack of Correlation with the Decidual Immune Cell Profile
Authors: PLEVYAK MICHAEL , HANNA NAZEEH , MAYER SANDRA , MURPHY SHAUN , PINAR HALIT , FAST LOREN , EKERFELT CHRISTINA , ERNERUDH JAN , BERG GORAN , MATTHIESEN LEIF , SHARMA SURENDRA
Published: March 2003

PROBLEM: To determine if first trimester missed abortion decidua is characterized by an altered immune cell profile and/or a modified interleukin (IL)‐10 and interferon (IFN)‐γ production pattern compared with decidua from elective termination.

METHOD OF STUDY: Flow cytometry and immunohistochemistry techniques were used to determine the decidual immune cell phenotypic profile and production pattern of IL‐10 and IFN‐γ in cases of elective termination (n=14) and missed abortion (n=12).

RESULTS: Both groups had a similar proportion of CD56+ CD16–, CD56+ CD16+, CD19+, CD3+, CD4+, CD8+, αβ T cells and γδ T cells. The majority of αβ and γδ positive T cells in both groups coexpressed the natural killer (NK) cell marker CD56, but lacked cell surface expression of CD3. Diminished decidual IL‐10 staining was noted in 7/10 missed abortion cases compared with none of the elective termination cases (n=12) (P=0.007). A uniform decidual IFN‐γ staining pattern was observed in both groups.

CONCLUSION: Decreased IL‐10 production coupled with a sustained IFN‐γ presence noted in missed abortion compared with elective termination cases suggest that these cytokines may be important determinants in pregnancy outcome. In contrast, differences in the proportion of immune cells between both groups may not be a critical factor in early pregnancy loss. In normal pregnancy, decidual αβ and γδ positive T cells with reduced CD3 on their cell surface may be intrinsically restricted in T‐cell receptor (TCR)‐mediated activation.
Distinct Expression Pattern of Cytokines in Semen of Men with Genital Infection and Oligo‐Terato‐Asthenozoospermia
Authors: Matalliotakis Ioannis , Arici Aydin , Goumenou Anastasia , Koumantakis Georgios , Selam Belgin , Matalliotakis Georgios , Koumantakis Evgenios
Published: March 2003

PROBLEM: The objective of this study was to evaluate the possible relevance of cytokines in seminal plasma (SP) of patients with accessory gland infection and oligo‐terato‐asthenozoospermia.

METHOD OF STUDY: Semen samples were obtained by masturbation from 90 men and were examined for the presence of interleukin (IL)‐2, IL‐6, IL‐8, IL‐11 and soluble CD23 (sCD23) by enzyme‐linked immunosorbent assay. Five groups were included: (1) fertile men (n=20), (2) infertile men with varicocele and oligo‐terato‐asthenozoospermia (V‐OTA, n=20), (3) infertile men with genital infection and OTA (INF‐OTA, n=20), (4) infertile men with idiopathic testicular lesion and OTA (ITL‐OTA, n=20) and (5) infertile men with azoospermia (AZOO, n=10). RESULTS:  We found that the mean level of IL‐2 was higher in SP from infertile men compared with SP from fertile men (P < 0.05). Mean levels of IL‐6, IL‐8, IL‐11 in SP of INF‐OTA were higher than that of all other groups (P < 0.05, P < 0.05, P < 0.001, respectively). However, no significant differences could be detected between other groups. A significant increase was noted in sCD23 levels in SP from men with ITL‐OTA compared with all other groups (P < 0.01). We have not observed any correlations between IL‐2, IL‐6, IL‐8, IL‐11 and sCD23 levels in SP and semen parameters. Spearman's correlation coefficient revealed that there was a significant association between IL‐6, IL‐8, IL‐11 levels in men with INF‐OTA.

CONCLUSION: The measurement of each cytokine separately in the SP of men with INF‐OTA, in spite of the existing significant differences, does not have a diagnostic value in male infertility. However, a combined determination of IL‐6, IL‐8, IL‐11 in the SP of men with genital infection and oligo‐terato‐asthenozoospermia may provide clinically useful information for the diagnosis of male accessory gland infection.
Transforming Growth Factors β1, β2 and β3 and their Receptors are Differentially Expressed in Human Peritoneal Fibroblasts in Response to Hypoxia
Authors: Saed Ghassan M. , Collins Karen L. , Diamond Michael P.
Published: March 2003

Saed GM, Collins KL, Diamond MP. Transforming growth factors β1, β2, and β3 and their receptors are differentially expressed in human peritoneal fibroblasts in response to hypoxia. AJRI 2002; 48: 387–393 © Blackwell Munksgaard, 2002

PROBLEM: Little is known about the role of peritoneal fibroblasts in adhesion formation. This study determines the effect of hypoxia and transforming growth factor (TGF)‐β1 treatment on the expression of TGF‐β1–3 and TGF‐βI and βII receptors in human peritoneal fibroblasts (HPF). TGF‐β isoforms and their receptors have been implicated as mediators of the healing process and adhesion development.

METHODS: HPF were cultured under normal and hypoxic condition, and treated with and without (1 ng/mL) TGF‐β1 for 24 hr. Total RNA from each group was subjected to multiplex reverse transcriptase‐polymerase chain reaction (RT/PCR) to quantitate TGF‐β1–3 and TGF‐βI and βII receptors messenger RNA (mRNA) levels.

RESULTS: Hypoxia resulted in a significant increase in TGF‐β1 (26%; P < 0.05), TGF‐βIR (34%; P < 0.05) and TGF‐βIIR (29%; P < 0.05) mRNA levels, with no effect on TGF‐β2 or β3. TGF‐β1 treatment resulted in a significant increase in TGF‐β1 (35%; P < 0.05), but a decrease in TGF‐β2 (22%; P < 0.05) and no effect on TGF‐β3, TGF‐βIR or TGF‐βΙIR. Combined treatment of hypoxia and TGF‐β1 caused a significant increase in TGF‐β1 (37%; P < 0.05), TGF‐β2 (12%; P < 0.05), TGF‐βIR (32%; P < 0.05) and TGF‐βIIR (34%; P < 0.05). There is no significant change in the mRNA levels of TGF‐β3 in any of the treatments.

CONCLUSION: Hypoxia and TGF‐β1 treatments of cultured HPF modulate the expression of TGF‐β1, β2 and β3 and their receptors βIR and βIIR by increasing the ratio of TGF‐β1 and β2 to β3, thus favoring the development of peritoneal adhesion.
Interleukin Regulation of Asymmetric Antibody Synthesized by Isolated Placental B Cells
Authors: CANELLADA A. , FÄRBER A. , ZENCLUSSEN A.C. , GENTILE T. , DOKMETJIAN J. , KEIL A. , BLOIS S. , MIRANDA S. , BEROD L. , GUTIÉRREZ G. , MARKERT U.R. , MARGNI R.A.
Published: March 2003

Canellada A, Färber A, Zenclussen AC, Gentile T, Dokmetjian J, Keil A, Blois S, Miranda S, Berod L, Gutiérrez G, Markert UR, Margni RA. Interleukin regulation of asymmetric antibody synthesized by isolated placental B cells. AJRI 2002; 48: 275–282 © Blackwell Munksgaard, 2002

PROBLEM: Protecting antibodies against trophoblast surface molecules were previously described. Here we analysed the synthesis of asymmetric IgG by placental B‐lymphocytes.

METHOD OF STUDY: B cells were isolated from human term placenta and cord blood, stimulated with anti‐CD40 IgG and cocultured with transfected Fcγ R‐expressing mice Ltk‐fibroblast. Interleukin‐4, IL‐6, IL‐10, IL‐11 and IL‐13 were added to cultures for 14 days. Asymmetric IgG were assessed in culture supernatants by concanavalin A (Con A) fixation and enzyme‐linked immunosorbent assay.

RESULTS: When IL‐6 was added to the cultures, the percentages of asymmetric IgG synthesized by placental B cells were: IL‐6: 29 ± 10; IL‐6 + IL‐10: 24 ± 7; IL‐4 + IL‐10 + IL‐6: 38 ± 9. The last combination induced the highest increase in the asymmetric IgG synthesis as compared with control (19 ± 10%, P < 0.05). Additionally, placental B cells synthesized more asymmetric IgG than umbilical cord blood B‐lymphocytes (P=0.0015).

CONCLUSIONS: Isolated placental B‐lymphocytes synthesized asymmetric IgG in response to Th2 interleukins, more notably IL‐6 in combination with IL‐4 and IL‐10. The in vitro increase of protective asymmetric IgG synthesis in response to Th2‐cytokines support the hypothesis that a local Th2‐switch is beneficial for pregnancy outcome.