bluuebunny/crossref_metadata_2025 huggingface dataset:
Therapeutic interventions in mice with chronic proliferative dermatitis (cpdm/cpdm)
Authors: Gijbels M. J. J. , Elliott G. R. , HogenEsch H. , Zurcher C. , Van Den Hoven A. , Bruijnzeel P. L. B.
Published: March 2003

Abstract: Chronic proliferative dermatitis (cpd) is a spontaneous mutation in C57BL/Ka mice (
cpdm/cpdm). The dermatitis is characterized by redness, hairloss, scaling, pruritus and histologically by epithelial hyperproliferation, infiltration of eosinophils, macrophages and mast cells. Lesions similar to those in the skin occur in the esophagus and forestomach. In this paper, we describe the effect of drug treatments directed against epidermal hyperproliferation (calcipotriene and etretinate), against inflammation (corticosteroids and dapsone) and against pruritus (loratidine and capsaicin). The criteria used to objectively estimate the effect of the treatment were 1) macroscopic evaluation of the lesions (cpd score), 2) degree of epithelial hyperproliferation assessed by
BrdU incorporation and epithelial thickness, and 3) microscopic evaluation of the inflammatory cells in the skin samples. Treatment of the cpdm/cpdm mice with calcipotriene (5 μg/day for 3 weeks) inhibited epidermal proliferation and the number of eosinophils. Systemic etretinate treatment (30 μg/g/day for 3 weeks) was not very effective. Topical corticosteroids (0.05 μg/day, for 3 weeks) exerted a therapeutic effect on the hyperproliferation and the number of eosinophils. Oral dapsone treatment (34 μg/g/day, for 5 weeks) reduced the BrdU incorporation in the skin and the epithelial thickness in the esophagus. The anti‐histamine loratidine (orally, 1.7 μg/g/day, for 4 weeks) reduced the severity of the lesions macroscopically, probably by suppressing the pruritus. Capsaicin (topically, 30 mM, for 5 weeks) also reduced the severity of the macroscopic observable lesions. Moreover, capsaicin reduced the dorsal and ventral epidermal thickness. The results from this and previous studies indicate that steroids (topically and systemically) and less strongly calcipotriene are the most effective treatments for the lesions observed in the cpdm/cpdm mice, since both hyperproliferation and the influx of eosinophils are reduced. Although the pathogenesis of the cpd lesions remains to be determined, our results indicate that the cpdm/cpdm mouse can be used to investigate new drugs for their possible application in chronic dermatitis.
Inhibition of intercellular adhesion molecule‐1 (ICAM‐1) expression in ultraviolet B‐irradiated human antigen‐presenting cells is restored after repair of cyclobutane pyrimidine dimers
Authors: Grewe M. , Stege H. , Vink A. , Klammer M. , Ruzicka T. , Roza L. , Krutmann J.
Published: March 2003

Abstract: The present study assessed the molecular mechanism underlying ultraviolet (UV) B radiation‐induced inhibition of the expression of the adhesion molecule ICAM‐1 in human antigen‐presenting cells (APC). UVB radiation‐induced inhibition of ICAM‐1 expression in human peripheral blood monocytes was associated with the generation of cyclobutane pyrimidine dimers (CPD). CPD were reduced by 60% after treatment with liposomal packed photolyase, an enzyme which removes CPD after absorption of photoreactivating light. Although incomplete, reduction of CPD was associated with complete restoration of ICAM‐1 expression at the mRNA and protein level. Neither reduction of CPD level nor restoration of ICAM‐1 expression were observed, if monocytes were treated with empty liposomes, or if they were irradiated with photoreactivating light prior to application of photolyase. DNA damage might also induce soluble mediators capable of autocrine inhibition of ICAM‐1 expression. UVB irradiation of monocytes did not induce IL‐10 production, but resulted in release of prostaglandin (PG) E2. Treatment of unirradiated monocytes with PGE2 completely inhibited ICAM‐1 expression, thus mimicking the UVB effect. Inhibition of monocytic PGE2 production by indomethacin, however, did not restore ICAM‐1 expression. These results suggest that formation of CPD is necessary and sufficient for UVB radiation‐induced inhibition of ICAM‐1 expression. In contrast, PGE2 might serve a paracrine role in UVB radiation‐induced immunosuppression.
Interspecies conservation and differential expression of mouse desmoglein gene family
Authors: Mahoney My˜ G. , Simpson Anisha , Aho Sirpa , Uitto Jouni , Pulkkinen Leena
Published: March 2003

Abstract: Epithelial cell adhesion is mediated by intercellular junctions, called desmosomes. Desmogleins (Dsg; Dsg1, Dsg2 and Dsg3) are calcium‐dependent transmembrane adhesion components of the desmosomes. While Dsg1 and Dsg3 are mainly restricted to stratified squamous epithelia, Dsg2 is expressed in essentially all desmosome‐containing epithelia. In the epidermis, Dsg2 and Dsg3 are expressed in the basal keratinocytes while Dsg1 is expressed throughout the upper differentiating cell layers. To date, in mouse, only Dsg3 has been characterized by molecular cloning. In this study, we have cloned and characterized the mouse Dsg1 and Dsg2 genes. The full‐length mouse Dsg1 cDNA (5.5 kb) contains an open reading frame (ORF) of 3171 bp encoding a precursor protein of 1057 amino acids. The Dsg2 cDNA (6.3 kb) has an ORF of 3366 bp coding for a precursor protein of 1122 amino acids. Mouse Dsg2 protein shares 76% identity with human DSG2 but only 26% and 33% identity with mouse Dsg1 and Dsg3, respectively. Analysis of intron/exon organization of the desmoglein genes revealed significant conservation. However, the mRNA expression patterns of these desmogleins during mouse embryonic development and in various adult tissues are variable. While Dsg2 and Dsg3 are expressed in all developmental stages, Dsg1 expression is delayed until day 15 of mouse embryos. In adult mouse tissues, Dsg2 is widely expressed while the expression of Dsg1 and Dsg3 is restricted to select tissues. In summary, while desmogleins share high homology at both the gene and protein level, their expression is spatially and temporally regulated, potentially contributing to their significant role in cell–cell adhesion during development.
Topical retinoic acid alters the expression of cellular retinoic acid‐binding protein‐I and cellular retinoic acid‐binding protein‐II in non‐lesional but not lesional psoriatic skin
Authors: Karlsson T. , Virtanen M. , Sirsjö A. , Rollman O. , Vahlquist A. , Törmä H.
Published: March 2003

Abstract: Therapeutic retinoids have profound effects on psoriatic skin pathology but their interactions with various retinoid‐binding proteins in lesional vs non‐lesional skin have not been investigated. Using quantitative real‐time PCR the mRNA expression of cellular retinol‐binding protein I (CRBPI) and retinoic acid‐binding protein I/II (CRABPI/CRABPII) was studied in psoriatic and healthy control (=normal) skin after 4 days of occlusive RA/vehicle treatment (n=6). Untreated psoriatic lesions showed a markedly elevated CRABPII/CRABPI ratio, while the CRBPI level was reduced in lesional and non‐lesional skin as compared to normal skin. In RA‐treated normal and non‐lesional skin, the mRNA expression of CRBPI was unaltered while that of CRABPI and CRABPII was reduced by ≈80% and increased ≈5‐fold, respectively, as compared to vehicle‐treated skin. In contrast, lesional skin exposed to RA showed an almost 90% increase in CRBPI transcripts but unaltered expression of CRABPI and CRABPII, yet, the mRNA expression of several inflammatory mediators, e.g. inducible nitric oxide synthase, interferon‐γ and interleukin‐1β, was clearly reduced. Immunohistochemistry localized CRABPII to suprabasal keratinocytes in normal skin and revealed markedly elevated levels in lesional skin. RA treatment induced CRABPII protein expression in normal and non‐lesional skin, to similar levels as in untreated lesions. The results indicate that the effects of RA differ in normal/non‐lesional psoriatic skin and lesional skin. Whether the high expression of CRABPII in psoriatic skin lesions is due to increased amounts of endogenous retinoids in lesional skin or reflects an abnormal regulation of the CRABPII gene in psoriasis remains to be studied.
Decreased IL‐10 production by psoriatic peripheral blood mononuclear cells stimulated with streptococcal superantigen
Authors: Aiba Setsuya , Uddin Zia , Nakagawa Satoshi , Sugawara Syunji , Rikiishi Hidemi , Kumagai Katsuo , Tagami Hachiro
Published: March 2003

Abstract:  The strong association of acute guttate psoriasis and streptococcal throat infection has suggested a role for streptococcal antigens in the pathogenesis of psoriasis. We have reported that psoriatic peripheral blood mononuclear cells (PBMCs) showed significantly lower responses to cytoplasmic membrane‐associated protein (CAP) isolated from group A β‐hemolytic streptococci, a kind of streptococcal superantigen. The objectives were to evaluate the abnormal cytokine production by psoriatic PBMCs to streptococcal superantigen, CAP. We compared the production of four different cytokines, i.e. IL‐4, IL‐5, IL‐10, and IFN‐γ, by PBMCs between psoriatic patients and healthy controls after stimulation with CAP or two different staphylococcal superantigens, staphylococcal enterotoxin A (SEA) or E (SEE). When PBMCs were stimulated with CAP, the production of IL‐10 was significantly lower by psoriatic PBMCs than by those from healthy controls, whereas those of IL‐4, IL‐5, or IFN‐γ were not different between the two groups. Such a significant decrease in IL‐10 production by psoriatic PBMCs was not observed when they were stimulated with staphylococcal superantigens. Flow cytometric analysis of intracytoplasmic IL‐10 demonstrated defective IL‐10 production by psoriatic PBMCs in both CD3+ T cells and CD14+ monocytes. There was a significant positive correlation between IFN‐γ production by PBMCs and the proliferation of Vβ8+ T cells preferentially stimulated by CAP. These data demonstrating the defective IL‐10 production by psoriatic PBMCs stimulated with streptococcal superantigen seem to explain why only psoriatic patients evolve sustained and Th‐1 deviated skin lesions after streptococcal upper respiratory infection.
Dual role of protein kinase C on mitogen‐activated protein kinase activation and human keratinocyte proliferation
Authors: Praskova M. , Kalenderova S. , Miteva L. , Poumay Y. , Mitev V.
Published: March 2003

Abstract: Subconfluent normal human keratinocytes exhibit autonomous (autocrine growth factor driven) proliferation and express the specific markers for keratinocyte proliferation K#5 and K#14. In keratinocyte autocrine culture, the exogenously added epidermal growth factor (EGF) has no effect on cell proliferation and mitogen‐activated protein kinase (MAPK) activity. PD98059 inhibits MAPK pathway and autocrine keratinocyte proliferation. Staurosporine and Gö6976 strongly inhibit autonomous keratinocyte proliferation. In contrast, Gö6983 (which does not inhibit PKCµ) inhibits only 20% of autocrine keratinocyte proliferation. Staurosporine inhibits MAPK activity, whereas Gö6976 and Gö6983 strongly increase it. We have concluded that MAPK, PKCµ and probably PKCα take part in autocrine keratinocyte proliferation. The effect of Gö6976 and Gö6983 on MAPK activity could be explained by the inhibition of PKC‐dependent MAPK‐phosphatase expression. The effect of staurosporine could be explained by its paradoxical action (activation) on protein kinase C (PKC) in keratinocytes.
Organotypic keratinocyte coculture using normal human serum:  an immunomorphological study at light and electron microscopic levels
Authors: Hinterhuber Gabriele , Marquardt Yvonne , Diem Edgar , Rappersberger Klemens , Wolff Klaus , Foedinger Dagmar
Published: March 2003

Abstract: Organotypic human skin equivalents of keratinocytes and fibroblasts embedded in collagen matrix have been the subject of studies dealing with various culture conditions. Development of standardized living skin equivalents using defined culture media containing respective supplements can provide important instruments of investigation in skin biology. In addition, tissue engineering has created human skin substitutes for treatment of acute and chronic wounds. In our study, we generate a modified organotypic human skin equivalent using normal human serum instead of fetal calf serum (FCS). This living skin equivalent shows regular stratification of the epidermis and the dermal–epidermal junction zone at the light and electron microscopic level after 1 and 3 weeks of coculture. Indirect immunofluorescence reveals regular expression of differentiation antigens and the major structural proteins collagen IV, laminin 5 and the integrin chains alpha 6 and beta 4 at the dermo–epidermal junction zone. Immunoelectron microscopy demonstrates expression of collagen IV, alpha 6 and beta 4 integrin after 1 and 3 weeks of coculture. This organotypic skin model could be the basis for autologous skin grafting for acute or chronic wounds using autologous serum as well as patients' keratinocytes and fibroblasts, thus minimizing the risk of transmitting infectious agents.
Mapping of the associated phenotype of an absent granular layer in ichthyosis vulgaris to the epidermal differentiation complex on chromosome 1*
Authors: Compton John G. , DiGiovanna John J. , Johnston Kay A. , Fleckman Philip , Bale Sherri J.
Published: March 2003

Abstract: Ichthyosis vulgaris (IV) is a mild to severe scaling disorder of uncertain etiology estimated to affect as many as 1 : 250 in the population. Family studies have shown that in many cases IV follows an autosomal dominant inheritance pattern, but gene mapping studies have not been reported. To investigate the genetic basis for inherited IV, we have performed gene linkage studies in two multigenerational families where affected individuals have clinical features of IV but distinct histological features. The epidermis in this disorder characteristically displays non‐specific orthohyperkeratosis. Notably, a subset of IV patients with a reduced or absent granular epidermal layer (AGL) have been reported, and decreased filaggrin levels have been described in others. The prominent role of profilaggrin in human keratohyalin suggests that defects in the gene for profilaggrin (FLG), its processing of profillagrin to filaggrin, or a gene involved in profilaggrin regulation may underlie or modify the pathology in IV. Family 1 had seven individuals with IV, severe heat intolerance and epidermis with 1–3 granular layers (consistent with normal epidermal histology). Ichthyosis vulgaris in this family did not segregate with FLG or other genes in the epidermal differentiation complex. In contrast, five of the six IV patients in Family 2, all siblings, had epidermis with no granular layer. Significant evidence was obtained for linkage of IV with the associated AGL phenotype to the epidermal differentiation complex (which includes FLG) assuming either a recessive (max Lod 3.4) or dominant (max Lod 3.6) inheritance model. Sequence analysis of FLG did not reveal a mutation in the amino or carboxyl terminal portions of the coding sequence adjacent to filaggrin repeats. The AGL may represent an endophenotype for IV, and the presence of a modifier of IV pathology at this locus is discussed.
Procyanidin B‐3, isolated from barley and identified as a hair‐growth stimulant, has the potential to counteract inhibitory regulation by TGF‐β1
Authors: Kamimura Ayako , Takahashi Tomoya
Published: March 2003

Abstract: With the aim of identifying natural products, which possess hair‐growing activity, we examined more than 1000 plant extracts with respect to their growth‐promoting effects on hair epithelial cells. We discovered intensive growth‐promoting activity, about 140% relative to controls, in barley extract. Our strategy for identifying active compounds in barley extract involved subjecting it to column chromatography using HP‐20 resin columns, an LH‐20 resin column, and preparative high‐performance liquid chromatography (HPLC) using an ODS column. The 60% (v/v) aqueous methanol eluted fraction from the HP‐20 column and the 75% (v/v) aqueous methanol eluted fraction from the subsequent LH‐20 column showed high hair‐growing activity in vivo. We isolated two major substances from the LH‐20 active fraction using preparative HPLC. By means of mass spectrometry, 1H‐NMR, and 13C‐NMR analyses, one substance was revealed to be procyanidin B‐3 and the other substance was identified as (+)‐catechin. Purified procyanidin B‐3 showed high hair‐growing activity in the form of in vitro hair epithelial cell growth‐promoting activity and in vivo anagen‐inducing activity; however (+)‐catechin showed no hair‐growing activity. For the purpose of examining the hair‐growing mechanisms of procyanidin B‐3, we examined its relationship to the TGF‐β signal pathway, which is known to be a regulator of catagen induction. Addition of TGF‐β1 to hair epithelial cell cultures dose‐dependently decreased the cell growth, and addition of procyanidin B‐3 to the culture neutralized the growth‐inhibiting effect of TGF‐β1. From these results, it is concluded that procyanidin B‐3 can directly promote hair epithelial cell growth in vitro, has the potential to counteract the growth‐inhibiting effect caused by TGF‐β1 in vitro, and has potential to stimulate anagen induction in vivo.
Growth and mortality of prairie stream fishes: relations with fish community and instream habitat characteristics
Authors: Quist M. C. , Guy C. S.
Published: March 2003

Abstract – Few studies have been conducted to describe the age structure, growth rates and mortality of fishes in small stream ecosystems. The purpose of this study was therefore to determine age structure, growth rates and mortality (i.e., total annual mortality and, age‐specific mortality) of central stonerollers Campostoma anomalum, creek chubs Semotilus atromaculatus, red shiners Cyprinella lutrensis and green sunfish Lepomis cyanellus from 13 streams on Fort Riley Military Reservation, Kansas, using incremental growth analysis. Further, we were interested in determining the influence of fish community and instream habitat characteristics on growth rates. The age structure of central stonerollers, creek chubs, and red shiners was dominated by young individuals (i.e., less than age 2); however, over 60% of the green sunfish were age 2 to age 4. Mean total annual mortality was >60% for cyprinids and averaged approximately 44% for green sunfish. The age‐specific mortality of central stonerollers and red shiners was generally less than 45% between age 0 and 1 and increased to over 85% for fishes greater than age 1. Fish community characteristics (e.g., catch per unit effort of trophic guilds) and chemical habitat (e.g., total phosphorous) were not related to growth rates (P>0.05). Growth of central stonerollers was not significantly correlated with physical habitat (P>0.05). However, the growth increments of creek chubs, red shiners, and green sunfish were related to the amount of woody debris (e.g., total woody debris, log complex habitat; r>0.60; P≤0.05). The results of this study provide important information on the population dynamic rate functions of cyprinid and green sunfish populations in small prairie streams. Furthermore, these data suggest that woody debris is important habitat influencing growth of stream fishes.Note
Validation of the otoliths for age determination in Barbus sclateri (Günth.)
Authors: Escot C. , Granado‐Lorencio C.
Published: March 2003

Abstract – Otoliths were used to explore the timing of annulus formation in Barbus sclateri. Lapillus exhibited opaque and translucent zones whose combination was formed once a year. Opaque zones appeared formed from July and January, and the translucent ones from February to June. We describe a procedure to assess age by using the number of annulii, the condition of the otolith’s edge and the capture date. The use of otoliths for age determination in this cyprinid endemic to the Iberian Peninsula provides a useful alternative to the traditional methods of scale reading.Note
Microsatellite diversity assessment of brown trout (Salmo trutta) population structure indicate limited genetic impact of stocking in a Norwegian alpine lake
Authors: Heggenes J. , Røed K. H. , Høyheim B. , Rosef L.
Published: March 2003

Abstract – Releases of non‐native trout often result in introgression into natural populations and negative genetic effects. The causal ecological mechanisms for a wide range of reported outcomes are poorly understood. Brown trout population structure in an alpine lake with three major recruitment streams was assessed by analysis for eight DNA microsatellite markers and compared with the non‐native population. The lake is subject to a 40‐year recorded history of stocking with exogenous trout. No certain deviations from Hardy–Weinberg equilibrium were found. Tests for population differentiation and genetic distance indicated separate populations for all the sampled areas, and with the exogenous population as a cluster quite different from the others. Assignment tests indicated that only a small fraction of the fish sampled from the lake originated from the introduced trout strain (<3%). Wild discriminate, naturally reproducing populations characterize this alpine lake ecosystem, in spite of 40 years of stocking, which appears to have had a limited impact. It is unlikely that this population structure can ultimately be explained by trout movement patterns. Genetic analysis needs to be supplemented by studies of local life history strategies, to evaluate the relative importance of local adaptation versus random genetic differentiation, because implications for conservation and management are different.
Phenotypic plasticity for life history traits in a stream population of the threespine stickleback,Gasterosteus aculeatusL.
Authors: Baker J. A. , Foster S. A.
Published: March 2003

Abstract – The life history traits of breeding size, clutch size, egg size and relative clutch mass were examined for evidence of plasticity within a set of six annual samples of threespine stickleback,Gasterosteus aculeatusL., females from south‐central Alaska, USA. Three samples (from 1992, 1996 and 1999) were of stickleback native to, and living within, a stream environment, while three (from 1995, 1996 and 1999) represented fish living within a pond environment recently colonized by the stream stickleback. Significant differences between stream and pond fish were found for all four traits. For most traits, pond‐living females showed greater variation both across and within years than did stream‐living females. Although extremely rapid evolution within the pond, or genetic drift caused by low founding population size, could not be completely ruled out, trait changes across years in both environments were interpreted as representing adaptive plasticity.
Filler particle leachability of experimental dental composites
Authors: Söderholm Karl‐Johan M. , Yang Mark C. K. , Garcea Ileana
Published: March 2003

We studied the effect of matrix selection, filler composition, and filler silanization on filler leachability after storage in distilled water or artificial saliva. We evaluated 2 matrix systems, 2 filler systems and 2 silane treatment procedures, combined into 8 different dental composite materials. A total of 128 batches were made, and 2 specimens per batch were prepared. Of these 2 specimens per batch, one was stored in distilled water and the other in artificial saliva, both at 37°C. We transferred the specimens each 30th day during a 3‐yr period to new vials containing either freshly distilled water or newly mixed artificial saliva and analyzed the solutions the specimens had been stored in regarding Si, Ba and Al concentrations. The analyses revealed that storage solution, filler composition, and total time in the storage solution had strong effects on leachability. The average monthly leakage of the three elements was linear with time and higher in the artificial saliva. The Ba‐containing filler leached Si faster in artificial saliva than in distilled water, and roughly twice as much as the quartz filler. The storage effect approached an order of magnitude, while the filler effect was roughly a factor of two. Filler leaching was linear over time.
Quantitative image analysis of hyaluronan expression in human tooth germs
Authors: Felszeghy Szabolcs , Hyttinen Mika , Tammi Raija , Tammi Markku , Módis László
Published: March 2003

The expression of hyaluronan in human tooth germs was studied by using a biotinylated hyaluronan‐binding complex and quantitative digital image analysis. At the cap stage, dental papilla exhibited a moderate staining, while intense reaction was observed in the apical portion of presecretory ameloblasts, stellate reticulum, and in dental basement membrane. When the enamel and dentine matrices started to develop, a strong hyaluronan reaction was evident in the young enamel and the apical portion of secretory ameloblasts. No hyaluronan could be detected in the secretory ameloblasts and enamel matrix of the early (9‐wk‐old) post‐natal stage. It is concluded that hyaluronan may play a transitory role in the early phase of the development of the enamel matrix organization. A very weak signal was observed in the wall of dentin tubules, whereas the rest of the dentine matrix was not stained. The odontoblasts and the pulp were also moderately stained, and these reactions gradually decreased with age, suggesting that hyaluronan may also contribute to the development of dentine matrix and pulp.
Role of interleukin‐1 in induction of matrix metalloproteinases synthesized by rat temporomandibular joint chondrocytes and disc cells
Authors: Ijima Yoshihiro , Kobayashi Masaru , Kubota Eiro
Published: March 2003

To identify cartilage‐degrading enzymes and cell types that can be specifically induced by interleukin‐1 (IL‐1)α, we studied matrix metalloproteinase (MMP) activities of cultured rat temporomandibular joint (TMJ) chondrocytes and disc cells. The cells were isolated from TMJs pre‐injected with normal physiological saline (CR) or recombinant human IL‐1α (AR). MMP activities in the conditioned media were assayed by gelatin enzymography, and they were identified by Western blot analyses. MMP mRNAs in these cells were also detected by RT‐PCR. IL‐1α significantly induced an increase of active MMP9 as well as pro‐ and active MMP3, but had no effect on the MMP2 activity in both types of cells. MMP3 and MMP9 mRNAs were also inducible in these cells by IL‐1α stimulation. Furthermore, disc cells were more susceptible to IL‐1α than chondrocytes. AR cells spontaneously produced the same MMPs in vitro as the CR cells synthesized under IL‐1α stimulation. The results indicate that MMP9 and MMP3 were predominantly produced by disc cells, and these may be considered to play a pivotal role in ECM degradation during pathological conditions of the TMJ, such as IL‐1‐induced TMJ arthritis.
Enhanced cyclooxygenase‐2 mRNA expression in human gingival fibroblasts induced by cell contact with human lymphocytes
Authors: Yucel‐Lindberg Tülay , Brunius Gustaf , Wondimu Biniyam , Andurén Ingegerd , Modéer Thomas
Published: March 2003

Prostaglandins, especially prostaglandin E2 (PGE2), play a crucial role in the pathogenesis of periodontal disease. We have previously reported that inflammatory mediators interleukin‐1 (IL‐1) and tumor necrosis factor α (TNFα) increase the production of PGE2 in human gingival fibroblasts. In this study, we investigated the effect of cell‐to‐cell interactions between gingival fibroblasts and lymphocytes on PGE2 production by using co‐culture technique. Cell‐to‐cell contact between gingival fibroblasts and lymphocytes synergistically enhanced the production of PGE2 in co‐cultures. In contrast to lymphocytes, the cyclooxygenase‐2 (COX‐2) mRNA expression in gingival fibroblasts was strongly enhanced following cell contact between gingival fibroblasts and lymphocytes. The level of COX‐1 mRNA expression, however, was not affected either in gingival fibroblasts or in lymphocytes by the interactions between fibroblasts and lymphocytes. The study demonstrates that cell contact between gingival fibroblasts and lymphocytes strongly stimulates PGE2 production partly due to enhanced COX‐2 mRNA expression in gingival fibroblasts. The cell‐to‐cell contact between gingival fibroblasts and lymphocytes should be considered as an important regulatory aspect for the enhancement of PGE2 in periodontal disease.
Osteoblastic activity of the rabbit temporomandibular joint during distraction osteogenesis assessed by [18F]fluoride positron emission tomography
Authors: Muhonen Arja , Peltomäki Timo , Knuuti Juhani , Raitakari Olli , Happonen Risto‐Pekka
Published: March 2003

The purpose of the study was to evaluate the effects of irradiation and hyperbaric oxygenation (HBO) on osteoblastic activity of the temporomandibular joint (TMJ) region during mandibular distraction osteogenesis. Unilateral distraction was performed on 19 rabbits, which were divided into five groups. One group served as a control group, while the others received either high‐ or low‐dose irradiation in the TMJ region before surgery. Some of the animals were also given HBO 18 times at 2.5 ATA×90 min preoperatively. Osteogenesis was assessed by [18F]fluoride positron emission tomography at the end of the distraction. Osteoblastic activity was higher on the distracted side in all groups, except in the high‐dose irradiated group without preceding HBO. HBO increased osteogenesis on both sides after radiotherapy. It is concluded that increased osteoblastic activity reflects increased pressure on the TMJ region of the distracted side, resulting from lengthening. It seems that more remodeling is required after irradiation than without preceding radiotherapy. After radiotherapy, HBO increased osteoblastic activity.
Nearly half of dental randomized controlled trials published in German are not included in Medline
Authors: Türp Jens C. , Schulte Jutta‐Maria , Antes Gerd
Published: March 2003

Randomized controlled trials (RCTs) are considered the most reliable type of clinical intervention studies. However, not all reports of RCTs are accessible in Medline. This can impede the validity of the results of systematic reviews. Ten German‐language dental journals were manually searched to locate reports of controlled clinical trials published between 1970 and 2000. The publication type was determined and compared with Medline. Of the 15 777 articles, 210 reports of RCTs and 410 articles of non‐randomized controlled clinical trials (CCTs) were identified. Only 56% of the RCTs and 75% of the CCTs are available in Medline. Of the 118 reports of RCTs registered in Medline, 15 are indexed with the correct Publication Type term. Our data suggest that (a) hand‐searching plays a valuable role in identifying reports of clinical dental trials, and (b) a literature search in Medline is likely to yield incomplete results.
Prenatal morphogenesis of the human mental foramen
Authors: Radlanski Ralf J. , Renz Herbert , Müller Ulrike , Schneider Richard S. , Marcucio Ralph S. , Helms Jill A.
Published: March 2003

The mental foramen, at first glance, merely looks like a hole where the mental nerve and the vascular bundle runs through. From a morphogenetic point of view, however, the mental foramen is a suitable model to study the development of a structure where different components are involved. To understand this developmental process, a three‐dimensional description at different developmental stages first has to be given. From histological serial sections of human embryos and fetuses, ranging in size from 19 to 117 mm crown rump length (CRL), three‐dimensional reconstructions of the foraminar regions were made. Outline and form of the developing foramen, size, course of the mental nerve and the adjacent blood vessels could be shown in detail. In this way, the formation of these structures became concrete in three dimensions. In the future, to understand the mechanisms regulating this complex system, where a nerve and blood vessels became successively surrounded by bone, molecular biological data have to be correlated with morphological findings.
Pulpal uptake of mercury from lined amalgam restorations in guinea pigs
Authors: Akyüz Serap , Çaglar Esber
Published: March 2003

The aim of the present study was to examine pulp in amalgam‐restored teeth of guinea pigs with respect to the presence of mercury, and to evaluate whether lining of cavities with resin modified glass ionomer cements had any effect on the penetration of mercury. Class V cavities were prepared in 63 incisor teeth of 21 guinea pigs. Three amalgam restorations were placed per animal, one without base liner and two with different resin‐modified glass ionomer cements (GC Lining LC and Ionoseal). Following observation periods of 1, 7 or 30 d, teeth were extracted and the mercury concentration in the pulp tissue was evaluated using atomic absorption spectrophotometry. It was found that resin‐modified glass ionomer cements significantly diminished the transport of mercury into the pulp between day 7 and day 30 after amalgam insertion.
Cytokine Network of Eutopic and Ectopic Endometrium in Women with Adenomyosis
Authors: SOTNIKOVA NATALIA , ANTSIFEROVA IOULIA , MALYSHKINA ANNA
Published: March 2003

PROBLEM: Recent studies showed the impairment of local cytokine balance in women with external endometriosis, but similar findings concerning direct production of cytokines by immunocompetent cells of women with adenomyosis are absent. In this context, investigation of the cytokine synthesis by mononuclear cells (MNCs) infiltrating eutopic and ectopic endometrium is of special interest.

METHOD OF STUDY: Concentration of interferon‐γ (IFNγ), interferon‐α (IFNα), tumour necrosis factor‐α (TNFα), interleukin‐1β (IL‐1β) and epidermal growth factor (EGF) in supernatants (SNs) of 24‐hr cultures of MNCs obtained from eutopic and ectopic endometrium of women with adenomyosis was determined by enzyme‐linked immunosorbent assay.

RESULTS: The levels of IFNγ, IFNα, TNFα, IL‐1β and EGF in SNs of eutopic endometrial MNCs of women with adenomyosis were significantly increased and the content of IL‐8 in SNs was reduced compared with that of the control figures. Ectopic MNCs of women with adenomyosis produced higher levels of IFNγ, IFNα and TNFα than the MNCs of normal endometrium. The production of IL‐1β, IL‐8 and EGF by ectopic endometrial MNCs was significantly reduced.

CONCLUSION: The results obtained indicate a significant role of local cytokine production impairment in the development of adenomyosis.
Some Thoughts on the Reproductive Autoimmune Failure Syndrome (RAFS) and Th‐1 Versus Th‐2 Immune Responses
Authors: GLEICHER NORBERT
Published: March 2003

Gleicher N. Some thoughts on the reproductive autoimmune failure syndrome (RAFS) and Th‐1 versus Th‐2 immune responses. AJRI 2002; 48:252–254 © Blackwell Munksgaard, 2002

PROBLEM: The concept of immunologically induced reproductive failure has remained very controversial. Especially the question whether abnormal immune function can cause infertility has recently been subject to strong disagreement. This opinion piece is an attempt to reconcile difference of opinion by integrating clinical data from the human experience with well‐established experimental models in animals.

METHOD OF STUDY: Review of published literature.

RESULTS: Experimental animal data, especially in the mouse, have suggested that normal pregnancy requires a switch to an immune response with T helper (Th)‐2 preponderance. Recent data from the human experience suggest that infertility, repeated pregnancy loss and certain complications of pregnancy may be the consequence of abnormal Th‐1 preponderance. In analogy, to animal models, immunologic reproductive failure in humans, may therefore not be the consequence of specific autoantibody abnormalities or natural killer (NK)‐cell abnormalities, but the reflection of a misdirection of a more broadly based immune response.

CONCLUSIONS: Under such a model, both diagnosis and treatment of immunologically induced forms of reproductive failure require re‐evaluation. Such a model, however, provides a satisfactory explanation for contradictory findings in the human experience that have led to strong disagreements among investigations.
Prostaglandin D2 and Reproduction
Authors: SAITO SHIGERU , TSUDA HIROSHI , MICHIMATA TOSHIHIKO
Published: March 2003

This review highlights recent studies investigating the role of prostaglandin (PG)D2 in reproduction. PGD2 induces sleep, allergic responses, inhibition of platelet aggregation, and relaxation of vascular and non‐vascular smooth muscle, and has some roles in reproduction.

Two types of PGD2 synthase are known. Lipocalin‐type PGD synthase is present in cerebrospinal fluid, seminal plasma and may play an important role in male reproduction. Another PGD synthase, hematopoietic PGD synthase is present in the spleen, fallopian tube, endometrial gland cells, extravillous trophoblasts and villous trophoblasts, and perhaps plays an important role in female reproduction. Recent studies demonstrate that PGD2 is probably involved in multiple aspects of inflammation through its dual receptor systems, DP and CRTH2. CRTH2 but not DP is a chemo‐attractant receptor for PGD2. Interestingly, CRTH2 is a most reliable marker for the detection of human T helper type 2 (Th2) and T cytotoxic type 2 (Tc2) cells, and the percentages of CRTH expressing CD4+‐T cells and CD8+‐T cells were significantly higher in the decidua especially at the implantation site, suggesting that Th2 and Tc2 cells recruit into the materno–fetal interface, in a PGD2‐mediated manner. PGD2 has a very unique effect to inhibit antigen presentation by inhibition of dendritic cell (DC) migration through DP but not CRTH2. PGD2 might appear to contribute to the maintenance of pregnancy by controlling the Th1/Th2 balance and antigen presentation by DCs through its dual receptor systems, CRTH2 and DP.
Absence of HLA‐G Expression in Macrophages of Human Decidua
Authors: SEDLMAYR PETER , MORALES PEDRO , TRUMMER SILVIA , WASCHER KARIN , AZZOLA DAGMAR , BLASCHITZ ASTRID , HAMMER ASTRID , WALCHER WOLFGANG , WINTERSTEIGER REINHOLD , DOHR GOTTFRIED
Published: March 2003

PROBLEM: Macrophages – together with natural killer (NK) cells – constitute the majority of bone marrow derived infiltrating cells in the decidua. As interferon‐γ (IFN‐γ), a cytokine produced by NK cells, has been reported to induce expression of human leukocyte antigen (HLA‐G) in monocytic cells, suggesting expression of HLA‐G on decidua macrophages potentially stimulated by IFN‐γ, the question arises whether decidua macrophages in normal pregnancy express HLA‐G.

METHOD OF STUDY: The study was based on immunohistochemistry and flow cytometry. In order to exclude that potentially elusive soluble HLA‐G was not detected by immunohistochemistry, we performed in addition RT–PCR of flow‐sorted decidua macrophages.

RESULTS: Our findings indicate that HLA‐G is not present on macrophages of first trimester or term decidua in either membrane‐bound or soluble form. Transcripts for soluble HLA‐G1 and ‐G2 were not detected.

CONCLUSIONS: We exclude a role of HLA‐G on the surface of decidua macrophages or of soluble HLA‐G1 or ‐G2 as a secretory product of decidua macrophages with regard to interaction with HLA‐G receptors present in or outside the decidua.
IL‐18 is Present at the Maternal–Fetal Interface and Enhances Cytotoxic Activity of Decidual Lymphocytes
Authors: TOKMADZ̆IĆ VLATKA S. , TSUJI YOSHIYUKI , BOGOVIĆ TATJANA , LAŠKARIN GORDANA , CUPURDIJA KRISTIJAN , Š TRBO NATAŠŠA , KOYAMA KOJI , OKAMURA HARUKI , PODACK ECKHARD R. , RUKAVINA DANIEL
Published: March 2003

Tokmadz̆ić VS, Tsuji Y, Bogović T, Laškarin G, Cupurdija K, Štrbo N, Koyama K, Okamura H, Podack ER, Rukavina D. IL‐18 is present at the maternal‐fetal interface and enhances cytotoxic activity of decidual lymphocytes. AJRI 2002; 48:191–200 © Blackwell Munksgaard, 2002

PROBLEM: Perforin expressing uterine natural killer (uNK) cells are under complex cytokine influence. The aim of the study was to investigate the presence and role of interleukin (IL)‐18 on NK cytolytic potential at maternal–fetal (M–F) interface.

METHOD OF STUDY: Peripheral blood cells and decidual tissue were obtained from elective pregnancy termination of normal human 6–10‐week‐old pregnancies. Perforin expression and cytolytic activity of peripheral blood (PBL) and decidual lymphocytes (DL) were analyzed by flow cytometry. IL‐18 positive decidual adherent cells (DAC) were detected by the same method. Interleukin‐18 and IL‐18 receptor (IL‐18R) expression on the trophoblastic cells was detected by immunohistology using biotinylated anti‐IL‐18 and IL‐18R monoclonal antibodies.

RESULTS: The IL‐18 added in a dose of 10 ng/mL up‐regulates perforin expression and cytolytic activity of DL. Simultaneous stimulation with IL‐18 and IL‐12 enhanced DL cytolytic activity, while IL‐18 combined with IL‐10 or IL‐15 did not show this effect. Cytolytic activity of PBL was up‐regulated by IL‐18 as well, and this effect was enhanced by the addition of IL‐12 and IL‐15. Interleukin‐18 did not affect perforin‐protein expression in cultured PBL. Approximately 20% of DAC were IL‐18 positive and these cells were mostly human leukocyte antigen (HLA)‐DR negative. IL‐18R positive cells were found on syncytiotrophoblast cell layer, interstitial tissue cells of villi and fetal blood cells. There was no detectable IL‐18 staining on trophoblast cell layer on villi, but strong staining of fetal blood cells in villous vessels.

CONCLUSION: These are first results showing IL‐18R expression, but not IL‐18 expression on villous trophoblastic cells, as well as enhancement of perforin expression and NK cytolytic potential of DL under the influence of IL‐18. IL‐18 in concert with other cytokines and hormones could play an important role in the regulation of cytolytic potential of first trimester pregnancy decidual and peripheral blood NK cells.
Modulation of T‐Cell CD3‐Zeta Chain Expression in Early Pregnancy*
Authors: EBLEN ABBY C. , GERÇEL ‐TAYLOR ÇIÇEK , NAKAJIMA STEVEN T. , TAYLOR DOUGLAS D.
Published: March 2003

PROBLEM: To assess the modulation of T‐cell CD3‐zeta chain expression by a factor in the sera of women, prior to egg retrieval and 14 days after an in vitro fertilization (IVF) and to delineate the mechanism of this modulation.

METHOD OF STUDY: In this prospective study, blood samples were obtained from 17 patients during an IVF cycle, prior to human chorionic gonadotropin (hCG), and 14 days after embryo return. Serum was incubated with cultured T‐lymphocytes (Jurkat cells) for 96 hr and expression of CD3‐zeta chain was evaluated.

RESULTS: Eight patients had a positive serum hCG titer 14 days after retrieval, while nine patients had a negative hCG titer. Serum from pregnant patients significantly suppressed CD3‐zeta chain expression as compared with their sample prior to retrieval (85.6 ± 6.2%), while subjects not becoming pregnant failed to suppress zeta expression (99.1 ± 0.9%, P < 0.0001).

CONCLUSION: A factor capable of suppressing TcR/CD3‐zeta expression can be detected in the sera of pregnant women 14 days after embryo retrieval. Loss of zeta chain was associated with the induction of apoptosis.
Sperm Immobilizing Antibodies in the Sera of Infertile Women Cause Low Fertilization Rates and Poor Embryo Quality In Vitro
Authors: TANEICHI AKIYO , SHIBAHARA HIROAKI , HIRANO YUKI , SUZUKI TATSUYA , OBARA HIROMI , FUJIWARA HIROYUKI , TAKAMIZAWA SATORU , SATO IKUO
Published: March 2003

PROBLEM: The effects of sperm immobilizing antibodies in the sera of infertile women on fertilization and embryo quality in vitro were investigated.

METHOD OF STUDY: Before the introduction of sperm immobilization test (SIT) as a routine test for female infertility, 85 oocytes were collected in nine in vitro fertilization (IVF) cycles from four infertile women who were afterward found having had sperm immobilizing antibodies in their sera and the oocytes were inseminated with swim‐up sperm in a medium containing the patient's serum. Fifty oocytes were collected in five IVF cycles from five infertile women possessing the antibodies in their sera and the oocytes were inseminated with swim‐up sperm in a medium supplemented with human serum albumin (HSA).

RESULTS: In the former group, 41 of 85 oocytes were fertilized, giving a fertilization rate of 48.2%. In the latter group, 43 of 50 oocytes were fertilized, giving a fertilization rate of 86.0%. There was a significant difference of the fertilization rate between the groups (P < 0.0001). Embryo quality was assessed by the Veeck's classification. The grade 1 and grade 2 embryos were considered good quality. Using this classification, 16 (39.0%) of 41 embryos incubated in the medium containing the patient's serum were good quality, while 34 (79.1%) of 43 embryos incubated in the medium supplemented with HSA were good quality. There was also a significant difference between the groups (P=0.0003).

CONCLUSIONS: These findings might indicate that sperm immobilizing antibodies in the sera of infertile women cause low fertilization rates and poor embryo quality in vitro. It is suggested that SIT in the sera of infertile women should be performed at least before proceeding IVF. The manipulation of gametes and embryos from patients having sperm immobilizing antibodies should be carefully carried out especially to avoid contaminating patient's serum and follicular fluid in the culture medium in order to have a better IVF result.
Regulation of the epididymal receptor tyrosine kinase ros by the protein tyrosine phosphatase SHP‐1
Authors: Pusch Rico , Böhmer Annette , Biskup Christoph , Benndorf Klaus , Lindenau Jörg , Böhmer Frank‐D.
Published: March 2003

The SH2 domain PTP SHP‐1 was recently identified as a potent negative regulator of the orphan receptor tyrosine kinase Ros, an important regulator of epidimys differentiation (Keilhack et al. J Cell Biol 2001; 152:325–334). Phosphorylated Ros strongly and directly associates with SHP‐1 in yeast‐two‐hybrid, GST pull‐down, and coimmunoprecipitation experiments. Catalytically inactive SHP‐1C455S exhibits greatly elevated binding to phosphorylated Ros. Direct Ros–SHP‐1 interaction is mediated by the SHP‐1 N‐terminal SH2 domain and Ros phosphotyrosine 2267. Overexpression of SHP‐1 results in Ros dephosphorylation and effectively down‐regulates Ros‐dependent proliferation and transformation. Elevated phosphorylation of Ros in ‘viable motheaten (me‐v)’ mice which, have strongly reduced SHP‐1 activity, suggests that Ros signaling is under control of SHP‐1 in vivo. Thus sterility of male me‐v mice seems to be related to dysregulation of Ros. A synthetic phosphopeptide derived from the Ros sequence around Y2267 potently activates recombinant SHP‐1 in vitro but is not a good substrate for SHP‐1. In contrast, phosphorylation sites in the activation loop of Ros are effectively dephosphorylated. Based on these observations we propose a mechanistic model of Ros–SHP‐1 interaction. Using fusion proteins of SHP‐1 variants and of Ros with GFP‐proteins of different spectral characteristics the interaction of Ros and SHP‐1 can be visualized in intact cells by different microscopic techniques.