bluuebunny/crossref_metadata_2025 huggingface dataset:
Flow cytometric measurement of reactive oxygen species production by normal and thalassaemic red blood cells
Authors: Amer Johnny , Goldfarb Ada , Fibach Eitan
Published: March 2003

Abstract: Reactive oxygen species (ROS) contribute to the pathogenesis of several hereditary disorders of red blood cells (RBCs), including thalassaemia. We report here on a modified flow cytometric method for measuring ROS in normal and thalassaemic RBCs. RBCs were incubated with 0.4 mM 2′,7′‐dichlorofluorescin diacetate (DCFH‐DA), then washed and further incubated either with or without 2 mM H2O2. Flow cytometric analysis showed that RBC fluorescence increased with time; it increased faster and reached higher intensity (by 10–30‐fold) in H2O2‐stimulated RBCs as compared to unstimulated RBCs. In both cases, the antioxidant N‐acetyl‐l‐cysteine reduced fluorescence, confirming previous reports that DCFH fluorescence is mediated by ROS. While the fluorescence of unstimulated RBCs increased with time, probably because of exposure to atmospheric oxygen, in H2O2‐stimulated RBCs fluorescence decreased after 30 min. The latter effect is most likely related to H2O2 decomposition by catalase as both sodium azide, an antimetabolite that inhibits catalase and low temperature increased the fluorescence of stimulated RBCs. Washing had a similar effect, suggesting that maintenance of the oxidised DCF requires a constant supply of ROS. We next studied RBCs of β‐thalassaemic patients. The results demonstrated a significantly higher ROS generation by stimulated and unstimulated thalassaemic RBCs compared to their normal counterparts. These results suggest that flow cytometry can be useful for measuring the ROS status of RBCs in various diseases and for studying chemical agents as antioxidants.
Severe IgA‐mediated auto‐immune haemolytic anaemia in a 48‐yr‐old woman
Authors: Bardill B. , Mengis C. , Tschopp M. , Wuillemin W. A.
Published: March 2003

Abstract: Auto‐immune haemolytic anaemia (AIHA) is characterised by haemolysis associated with the presence of the immunoglobulins IgG, IgM or IgA, and/or components of the complement system on the red cell membrane. The immunoglobulins react as auto‐antibodies against the red cell antigens of the patient. IgG antibodies and the complement component C3d can be detected by the direct antiglobulin test (DAT); however, IgM and particularly IgA antibodies may not necessarily be detected by the broad‐spectrum anti‐human‐globulin serum. We present the case of a 48‐yr‐old woman with severe AIHA. The initial polyspecific direct antiglobulin test (DAT), using a broad‐spectrum antiserum, was negative. Testing with monospecific antisera led to the diagnosis of AIHA due to warm‐acting auto‐antibodies solely of the IgA class, which is a very rare finding. As therapy with steroids alone did not lead to a lasting remission, splenectomy was performed 10 months after initial diagnosis. There has been no relapse of AIHA since, even after steroid medication was withdrawn and even though the monospecific IgA‐DAT has remained positive. This case demonstrates the importance of performing a monospecific antiglobulin test if there is a strong suspicion of AIHA in apparently ‘Coombs‐negative’ haemolytic anaemia. In AIHA caused by solely IgA antibodies, the polyspecific direct antiglobulin test may be negative or only weakly positive because of a limited content of anti‐IgA antibodies in the polyspecific anti‐human‐globulin serum. First‐line treatment of warm‐type AIHA is the administration of high‐dose glucocorticosteroids; splenectomy is indicated in steroid‐refractory patients.
p53 mutations as a marker of skin cancer risk: comparison of UVA and UVB effects
Authors: De Gruijl Frank R.
Published: March 2003

Abstract:The epidermis is excellently adapted to the sun's ultraviolet (UV) radiation. The p53 protein plays a crucial role in the orchestration of a cell's response to UV‐induced damage, and more specifically to DNA damage. This response appears to differ between differentiated (suprabasal) and undifferentiated (basal) epidermal cells. The latter are the most likely targets in UV carcinogenesis. The UVB‐related mutations inp53genes of human carcinomas from sun‐exposed skin indicate that rendering p53 dysfunctional is an important (early) step in the formation of these tumors. Experiments in hairless mice confirm this finding for UVB‐driven carcinogenesis, but not for UVA1‐(365‐nm)‐driven carcinogenesis. Microscopic clusters of preneoplastic cells overexpressing mutant p53 occur in chronically UVB‐exposed murine skin long before the ultimate carcinomas. The number of these clusters at a certain time‐point appears to be predictive of the tumor risk at latter time‐points. These UVB‐induced p53 clusters appear to be suitable surrogates of tumors in short‐term experiments.
Identification of a novel mutation in keratin 1 in a family with epidermolytic hyperkeratosis
Authors: Arin M. J. , Longley M. A. , Epstein E. H. , Rothnagel J. A. , Roop D. R.
Published: March 2003

Abstract: Epidermolytic hyperkeratosis (EHK) is a hereditary skin disorder typified by blistering due to cytolysis. One in 100,000 individuals is affected by this autosomal‐dominant disease. The onset of the disease phenotype is typically at birth. Histological and ultrastructural examination of the epidermis shows a thickened stratum corneum and tonofilament clumping around the nucleus of suprabasal keratinocytes. Linkage studies localized the disease genes on chromosomes 12q and 17q which contain the type II and type I keratin gene clusters. Recently, several point mutations in the genes encoding the suprabasal keratins, K1 and K10, have been reported in EHK patients. We have investigated a large kindred affected by EHK and identified a new point mutation in the 2B region of keratin 1 (I107T), resulting from a T to C transition in codon 478.
The 500 Dalton rule for the skin penetration of chemical compounds and drugs
Authors: Bos Jan D. , Meinardi Marcus M. H. M.
Published: March 2003

Abstract: Human skin has unique properties of which functioning as a physicochemical barrier is one of the most apparent. The human integument is able to resist the penetration of many molecules. However, especially smaller molecules can surpass transcutaneously. They are able to go by the corneal layer, which is thought to form the main deterrent. We argue that the molecular weight (MW) of a compound must be under 500 Dalton to allow skin absorption. Larger molecules cannot pass the corneal layer. Arguments for this “500 Dalton rule” are; 1) virtually all common contact allergens are under 500 Dalton, larger molecules are not known as contact sensitizers. They cannot penetrate and thus cannot act as allergens in man; 2) the most commonly used pharmacological agents applied in topical dermatotherapy are all under 500 Dalton; 3) all known topical drugs used in transdermal drug‐delivery systems are under 500 Dalton. In addition, clinical experience with topical agents such as cyclosporine, tacrolimus and ascomycins gives further arguments for the reality of the 500 Dalton rule. For pharmaceutical development purposes, it seems logical to restrict the development of new innovative compounds to a MW of under 500 Dalton, when topical dermatological therapy or percutaneous systemic therapy or vaccination is the objective.
What are the most promising strategies for the therapeutic immunomodulation of allergic diseases?Note
Authors: Tokura Y. , Röcken M. , Clark Richard A. F. , Maurer M. , Grabbe S. , Haliasos E. , Takigawa M. , Sinha A. A.
Published: March 2003

Abstract: Specific immunotherapy and other immunomodulatory strategies have long been a stronghold in the management of allergic diseases. In particular, “immunodeviation‐therapy” or “vaccination for allergies”, i.e. the redirection of Th2‐type immune responses towards a Th1‐response pattern, has become an ever more popular concept. The present feature of CONTROVERSIES complements our previous discussion of atopy (Röcken et al., Exp Dermatol 7: 97–104, 1998), and is dedicated to a critical analysis of the general problems and limitations one faces with the main immunomodulatory strategies traditionally considered in this context. We also explore alternative approaches that appear promising in order to achieve both a more effective and/or a more specific immunotherapy of allergic diseases. Given that the mast cell remains a key protagonist in the pathogenesis of allergic diseases finally, this feature examines how innovative, more selectively mast cell‐targeted strategies may be developed for the management of allergic diseases.
Proprotein convertase expression and localization in epidermis: evidence for multiple roles and substrates
Authors: Pearton David J. , Nirunsuksiri Wilas , Rehemtulla Alnawaz , Lewis S. Patrick , Presland Richard B. , Dale Beverly A.
Published: March 2003

Abstract: Specific proteolysis plays an important role in the terminal differentiation of keratinocytes in the epidermis and several types of proteases have been implicated in this process. The proprotein convertases (PCs) are a family of Ca2+‐dependent serine proteases involved in processing and activation of several types of substrates. In this study we examined the expression and some potential substrates of PCs in epidermis. Four PCs are expressed in epidermis: furin, PACE4, PC5/6 and PC7/8. Furin is detected in two forms, either with or without the transmembrane domain, suggesting occurrence of post‐translational cleavage to produce a soluble enzyme. In addition the furin active site has differential accessibility in the granular layer of the epidermis relative to the basal layer, whereas antibodies to the transmembrane domain stain both layers. These findings suggest that furin has access to different types of substrates in granular cells as opposed to basal cells. PC7/8, in contrast, is detected throughout the epidermis with antibodies to both the transmembrane and active site and no soluble form observed. A peptide PC inhibitor (dec‐RVKR‐CMK) inhibits cleavage of Notch‐1, a receptor important in cell fate determination that is found throughout the epidermis. Profilaggrin, found in the granular layer, is specifically cleaved by furin and PACE4 in vitro at a site between the amino terminus and the first filaggrin repeat. This work suggests that the PCs play multiple roles during epidermal differentiation.
Assessment of adhesion assays for use with keratinocytes
Authors: May A. L. , Wood F. M. , Stoner M. L.
Published: March 2003

Abstract: Cell attachment, as a biological process, is an important aspect with respect to graft survival and “take”. With the ever‐increasing use of cultured epithelial autografts for coverage and re‐epithelialization of wounds, the assessment of keratinocyte adhesion in vitro has become a more common requirement in studies involving extracellular matrix proteins and their receptor molecules. Cell adhesion has been well‐documented in immunological research, however keratinocyte adhesion has been investigated by manual counting (using methylene blue) or other less sensitive colorimetric methods. With the increase in number of fluorogenic probes available, their use as a sensitive alternative to radioactive labelling has been promoted in the literature. This study was carried out to investigate the possibility of using fluorescent probe 5,6‐carboxyfluorescein diacetate succidimyl ester to achieve a more standardized assay in the assessment of keratinocyte adhesion. Adhesion was assessed on extracellular matrix proteins such as fibronectin, collagen types I & IV and laminin. We concluded that the fluorescent probe might provide greater sensitivity in measuring adhesion, however it may be cytotoxic to keratinocytes. Pre‐labelling of keratinocytes may affect cellular functions such as adhesion and even proliferation and consequently the probe must be chosen with care.
Neuromediators and pigmentation
Authors: Abdel‐Malek Zalfa
Published: March 2003

FONDATION RENE TOURAINE POUR LA RECHERCHE EN DERMATOLOGIE

SCIENTIFIC MEETING 2001

Thursday 15th November 2001, Paris, France

Ministère de la Recherche et de la Technologie

Amphithéâtre Poincaré, 1, rue Descartes, F‐75005 Paris

***

NEUROPHYSIOLOGY OF THE SKIN

9h30–10h30 E. DUPIN Embryogenesis of neuro‐epithelial interactions

10h30–11h15 D. METZE Functional anatomy of the skin nervous system

11h15–12h00 T. LUGER Neuroimmunology

12h00     Winners of fellowships

12h30     Cocktail and Lunch

14h30–15h00 H. O. HANDWERKER Neuromediators, pruritus and pain

15h00–15h30 J. ANSEL Neuromediators and inflammation

15h30–16h00 Z. ABDEL‐MALEK Neuromediators and pigmentation

Break

16h30–17h00 R. PAUS Stress and skin

17h00–17h30 N. W. BUNNETT Pharmacological modulation of skin neuromediators (PAR II)

La Fondation René Touraine est une Fondation reconnue d’utilité publique.

Les Membres Fondateurs sont: les Laboratoires BAILLEUL, BERGADERM, BOOTS‐LUTSIA, CHANEL, PIERRE FABRE, FONDATION LEO, GALDERMA, LABORATOIRES SÉROBIOLOGIQUES, LA ROCHE POSAY, L’OREAL, L.V.M.H., ROCHE, SCHERING PLOUGH.

Inscription : Fondation René Touraine, Hôpital Saint‐Louis, Pavillon Bazin, 1, Avenue Claude Vellefaux – 75475 – Paris – France.

Tél : ++33 1 53 72 20 60 – Fax : ++33 1 53 72 20 61 – e‐mail: fond.r.touraine@chu-stlouis.fr
Assessment of contact allergen cross‐reactivity by retesting
Authors: Rustemeyer Thomas , De Groot Jan , Von Blomberg B. Mary E. , Bruynzeel Derk P. , Frosch Peter J. , Scheper Rik J.
Published: March 2003

Abstract: At former allergic contact dermatitis reaction sites retesting causes augmented hyper‐reactivity, characterized by an accelerated onset within a few hours. This expression of ‘local skin memory’ has been ascribed to locally persisting allergen‐specific effector/memory T cells. To verify this hypothesis, we investigated whether accelerated retest reactivity also occurs with cross‐reactive allergens. Guinea pigs were immunized with either or both 2,4‐dinitrochlorobenzene (DNCB) and 2‐hydroxyethyl methacrylate (HEMA), and primary skin tests to these and cross‐reactive methacrylic compounds were performed 12–21 days later. Subsequently, new skin tests were conducted 3 weeks later both at the former test (‘retest’) and contralateral, non‐pretreated test (‘control’) sites, and skin test readings started 2 h later. Retest reactivity was evaluated by comparing retest and contralateral control reactions. Both contact sensitizers, HEMA and DNCB, induced strong retest reactivity, peaking at 4–6 h. Fully allergen‐specific retest reactivity was observed when primary skin tests had been postponed until 21 days after immunization, most probably reflecting loss of accumulation of irrelevant allergen‐primed T cells at that time. As hypothesized, retesting with various methacrylate congeners at primary HEMA, but not DNCB, skin test sites showed early hyperreactivity strengths in line with those observed earlier in conventional cross‐reactivity studies. These results, therefore, support the view that local skin memory exhibits allergen specificity through residual allergen‐primed T cells. Because the retesting procedure is readily applicable in clinical practice, it provides a tool not only for confirmation of doubtful contact allergic skin reactions, but also for distinguishing between true cross‐reactivity and coincident multiple sensitization in man.
Improved detection of lacZ reporter gene expression in transgenic epithelia by immunofluorescence microscopy
Authors: Mahony Donna , Karunaratne Seetha , Rothnagel Joseph A.
Published: March 2003

Abstract: The bacterial lacZ gene is commonly used as a reporter for the in vivo analysis of gene regulation in transgenic mice. However, several laboratories have reported poor detection of β‐galactosidase (the lacZ gene product) using histochemical techniques, particularly in skin. Here we report the difficulties we encountered in assessing lacZ expression in transgenic keratinocytes using classic X‐gal histochemical protocols in tissues shown to express the transgene by mRNA in situ hybridization. We found that lacZ reporter gene expression could be reliably detected in frozen tissue sections by immunofluorescence analysis using a β‐galactosidase‐specific antibody. Moreover, we were able to localize both transgene and endogenous gene products simultaneously using double‐label immunofluorescence. Our results suggest that antibody detection of β‐galactosidase should be used to verify other assays of lacZ expression, particularly where low expression levels are suspected or patchy expression is observed.
Epidermal CD8+ T cells reactive with group A streptococcal antigens in chronic plaque psoriasis
Authors: Ovigne J.‐M. , Baker B. S. , Davison S. C. , Powles A. V. , Fry L.
Published: March 2003

Abstract: Chronic plaque psoriasis is a T cell mediated disease associated with group A streptococci (GAS). We have previously shown the presence of a psoriasis‐specific dermal Th1 subset that recognizes GAS antigens. To assess whether GAS‐reactive T cells are also present in lesional epidermis, fresh cell suspensions or T cell lines isolated from lesional epidermis of 33 psoriasis patients were stained for intracellular interferon‐γ after stimulation with GAS antigens. The patients were typed by PCR for HLA‐DR7 and HLA‐Cw6 expression. A subset of GAS‐reactive CD8+ T cells (2.4% ± 2.4) was found in 14/21 (67%) fresh cell suspensions. A smaller subset of GAS‐reactive CD4+ T cells (0.9% ± 0.9) was found in 13/21 (62%) fresh cell suspensions, which was expanded in the T cell lines. There was a significant inverse correlation between the proportions of GAS‐reactive CD4+ and CD8+ T cells in the fresh suspensions (r = −0.48, P = 0.0277). The presence of GAS‐reactive CD4+ or CD8+ T cells did not correlate with HLA‐DR7 or HLA‐Cw6 expression, respectively. This study has demonstrated GAS‐reactive CD8+, and to a lesser extent CD4+, T cell subsets in psoriatic epidermis and provides further evidence that GAS antigens may play a role in the pathogenesis of chronic plaque psoriasis.
Simple and rapid method to isolate and culture follicular papillae from human scalp hair follicles
Authors: Magerl Markus , Kauser Söbia , Paus Ralf , Tobin Desmond J.
Published: March 2003

Abstract: Study of the involvement of the hair follicle papilla in hair growth regulation was greatly facilitated by the isolation and cultivation of this tiny cluster of fibroblast‐like cells in the rat vibrissae and in the human hair follicle. While isolation of the hair follicle papilla from the former is relatively straightforward, the current method to isolate the much smaller human hair follicle requires significant skill. Thus, the routine initiation of primary cultures of human scalp hair follicle papilla cells requires significant training, time, and commitment.

In an attempt to simplify hair follicle papilla cell culture methodology for new laboratory personnel, we have made significant refinements to the current method. Our method requires only two simple manipulations to isolate hair follicle papilla from intact isolated hair follicles. This very rapid and easy method isolates clean and intact hair follicle papillae. Together with their attachment via scratching to the growth surface, the isolation and cultivation of this important hair follicle component can now be achieved easily by the laboratory newcomer. The method relies for its simplicity on the removal of the hair follicle papilla from the outside of the intact hair follicle rather than via internal manipulations from within the hair follicle.
Paternal germline mosaicism in Herlitz junctional epidermolysis bullosa
Authors: Cserhalmi‐Friedman Peter B. , Anyane‐Yeboa Kwame , Christiano Angela M.
Published: March 2003

Abstract: We studied a single patient with the lethal (Herlitz) type of junctional epidermolysis bullosa (H‐JEB). Screening for mutations in the LAMB3 gene in the patient revealed the previously described hotspot mutation R635X and a novel one basepair deletion in exon 10. The single basepair deletion 1094delA could be detected in the clinically unaffected mother, while the nonsense mutation R635X could not be found in the peripheral blood DNA of either parent. After excluding non‐paternity by microsatellite analysis using random markers on chromosomes 3, 8 and 18, we determined that the mutation R635X in the proband was most likely the result of a de novo event or alternatively, germline mosaicism. The parents requested prenatal diagnosis for a second pregnancy, and while the maternal mutation 1094delA could not be detected in DNA from the fetus, unexpectedly, the mutation R635X was present in the chorionic villus DNA. These findings were most consistent with paternal germline mosaicism for the recessive mutation R635X. The results have had a significant impact on the genetic counseling in this family. To our knowledge, this study represents the first documented case of germline mosaicism in junctional epidermolysis bullosa, and serves as a reminder that germline mosaicism should be considered in cases in which a ‘new’ mutation is found in the offspring of a clinically and/or genetically unaffected parent.
Growth and diet of anadromous Arctic charr after their return to freshwater
Authors: Rikardsen A. H. , Amundsen P.‐A. , Bodin P. J.
Published: March 2003

Abstract –The post smolt of anadromous Arctic charr (Salvelinus alpinus) from Lake Storvatn, northern Norway (70°39′48″), continued to feed and grow after their return to freshwater in late summer, but in late autumn (October) both their growth rate and feeding intensity ceased along with the drop in water temperature. In contrast, adult, sexually mature anadromous charr did not appear to feed during their freshwater residency. The stomach fullness of post smolts were similar to that of resident Arctic charr during early autumn, and their diets were similar, being dominated by pelagic prey such as surface insects,Holopediumsp.,Bosminasp. and chironomid pupae. Later in the autumn, the resident charr also included some more benthic prey, in particular gastropods, whereas the post smolts continued to feed predominantly on zooplankton and surface insects, but with a lower stomach fullness than the resident fish. This suggests that after arrival in freshwater, the post smolts continue their pelagic feeding behaviour displayed during the sea‐migration.
Characterization of a family with dominant hypophosphatasia
Authors: Hu Jan C.‐C. , Plaetke Rosemary , Mornet Etienne , Zhang Chuhua , Sun Xiaoling , Thomas Huw F. , Simmer James P.
Published: March 2003

A kindred with dominant hypophosphatasia resulting from an alanine to threonine substitution at position 99 of the alkaline phosphatase protein is described. The clinical findings of individual members of the kindred were assessed by oral and physical examinations, or from the descriptions of multiple family members. The proband displayed enamel hypoplasia and premature loss of fully rooted primary anterior teeth, which were shown by histological examination to lack cementum. Serum alkaline phosphatase (ALP) and a vitamin B6 panel, and urine phosphoethanolamine (PEA) were measured on 21 family members. Based upon the clinical and laboratory tests, affected and unaffected status was assigned. Parametric linkage analysis of the kindred using different dominant models and frequency distributions for the disease allele and the mutation gave lodscores >4.2 and confirmed the strong linkage between the disease and the mutation. Assuming the defined mutation causes the disease, the reliability of clinical and laboratory tests is assessed.
The nanoleakage phenomenon: influence of different dentin bonding agents, thermocycling and etching time
Authors: Dörfer Christof E. , Staehle Hans J. , Wurst Marcus W. , Duschner Heinz , Pioch Thomas
Published: March 2003

“Nanoleakage” takes place within the hybrid layer zone of the dentin‐composite interface in spaces not occupied by polymerized resin. The purpose of this study was to quantify the amount of nanoleakage in specimens treated with one of six different bonding agents. For one agent, different etching times were used, and for two agents results after thermocycling were obtained. Standardized class V cavities were prepared in 165 extracted human molars with cervical margins located in dentin. After placement of the composite using bonding agents, the teeth were stored in a 1% rhodamin‐B‐isothiocyanate solution for 24 h at 20°C, embedded in methacrylate, and sectioned parallel to the long axis of the tooth. A confocal laser scanning microscope was used to visualize a layer 10 μm below the prepared surface of the section. The lengths of the penetrated pathways were measured, representing the amount of nanoleakage. In all materials tested, penetration pathways appeared within the hybrid layer in absence of gap formation. Penetration lengths of the tested materials were in a range from 69±24 μm to 469±333 μm. Thermocycling had no statistically significant influence, and etching for 15 s resulted in statistically significantly shorter penetration compared to longer etching.
Enamelin maps to human chromosome 4q21 within the autosomal dominant amelogenesis imperfecta locus
Authors: Dong J. , Gu T. T , Simmons D. , MacDougall M.
Published: March 2003

Amelogenesis imperfecta is a group of hereditary enamel defects. Of the autosomal dominant forms, only the local hypoplastic type has been mapped to human chromosome 4q13–4q21. Enamelin is a large enamel matrix protein secreted by ameloblasts. The purpose of this study was to determine the human chromosomal localization of enamelin to establish an association with various forms of amelogenesis imperfecta. Chromosomal mapping was performed by polymerase chain reaction (PCR) amplification using somatic hybrid and deletion/derivation cell line panels with an enamelin primer set based on 100% conserved regions between pig and mouse cDNAs. Sequence‐tagged site content mapping using eight markers within the critical local hypoplastic amelogenesis imperfecta region was then performed using an isolated human enamelin genomic BAC clone. The human enamelin amplicon was confirmed by DNA sequence analysis, revealing 81% and 73% identity to pig and mouse cDNAs, respectively. PCR amplification using a somatic cell hybrid panel placed enamelin on chromosome 4 with analysis of a regional chromosome 4 mapping panel refining the localization to 4q13.1–q21.23. An identified human enamelin BAC genomic clone was shown to contain markers D4S2604 and D4S2670, as well as the first exon of the human ameloblastin gene, placing enamelin in the critical amelogenesis imperfecta locus between markers HIS1 and D4S2604 at 4q21. Our results suggest that enamelin is a strong candidate gene for this disease. Furthermore, human 4q21 may contain a second cluster of enamel matrix genes located proximally to the identified cluster of dentin and bone genes.
Dental plaque mass and acid production activity of the microbiota on teeth
Authors: Borgström Margareta Krasse , Edwardsson Stig , Sullivan Åsa , Svensäter Gunnel
Published: March 2003

With the purpose of elucidating the effect of dental plaque thickness on the acid production activity in dental plaque, we studied the prevalence of acid anions in dental plaque of children harbouring different amount of plaque on their teeth. On two occasions, the occurence of plaque on the dentition was scored (Silness‐Löe's index), and plaque on available smooth surfaces was collected and the wet weight determined. On the first occasion, the amount of acid anions in resting plaque, and on the second occasion, the acid anions of sucrose‐exposed plaque were analysed with isotachophoresis. Mean value of plaque wet weights per individual dentition was 11.7±9.0 mg (first occasion) and 11.1±8.9 mg (second occasion). Dominating anions in resting plaque were acetate<propionate<lactate and in sucrose‐exposed plaque lactate<acetate<propionate. The microbial acid production activity (acid anion per mg wet plaque weight) decreased with increasing weights of the plaque mass. The findings illustrate the cellular glycolytic metabolism of plaque microorganisms in thin and thick plaque, being a consequence of the diverse enviromental condition existing in these ecosystems.
Does chemomechanical caries removal affect dentine adhesion?
Authors: Haak Rainer , Wicht Michael J. , Noack Michael J.
Published: March 2003

The purpose of this study was to determine whether shear bond strengths (SBS) to chemomechanically excavated dentine (Carisolv™) differed from the SBS after conventional caries removal (bur). The following adhesive systems were used: Dyract AP/NRC/Prime & Bond NT; Dyract AP/Prime & Bond NT; Tetric Ceram/Syntac SC; Tetric Ceram/Syntac SC (no etch)(TC 2); Definite/Etch & Prime 3.0; Definite/experimental. One‐hundred and twenty human molars with occlusal caries were assigned to 12 groups. Sequential caries removal was controlled with laser fluorescence. After water storage, the samples were tested in a single‐plane shear test assembly. The results demonstrated differences between the bonding systems, whereas the mode of caries removal had no consistent effect. If total etch technique was neglected, the results of the composite resin (TC 2) indicated a tendency to higher SBS in the Carisolv group (18.6±4.6 MPa) compared with conventional treatment (14.1±3.9 MPa). The present data demonstrate that chemomechanical caries removal has no adverse effect on bonding of modern adhesive systems to dentine. Smear layer‐dissolving or ‐modifying bonding systems could potentially benefit from chemomechanical pretreatment.
Actinobacillus actinomycetemcomitans‐induced expression of IL‐1α and IL‐1β in human gingival epithelial cells: role in IL‐8 expression
Authors: Sfakianakis Andreas , Barr Charles E. , Kreutzer Don L.
Published: March 2003

Gingival epithelial cells (GEC) are the first cells of the host that encounter the periodontal pathogens, and therefore their role in the initiation of the inflammatory response is critical. We aimed to: 1) characterize the expression of interleukin (IL)‐1α and IL‐1β in human gingiva and cultured GEC; 2) demonstrate the ability of A. actinomycetemcomitans extracts to upregulate IL‐1α, IL‐1β and IL‐8 expression in GEC in vitro; and 3) characterize the role of IL‐1α and IL‐1β in the induction of IL‐8 expression in GEC in vitro. Ten gingival biopsies (5 inflamed and 5 controls) and cultured GEC were examined for IL‐1α and IL‐1β using immunohistochemical techniques. GEC were also challenged with A. actinomycetemcomitans extracts or IL‐1, and secretion of IL‐1 and IL‐8 was determined by ELISA. In vivo, IL‐1α and IL‐1β were localized in the gingival epithelium and the infiltrating leukocytes. In vitro, A. actinomycetemcomitans extracts induced a time‐dependent expression of IL‐1α, IL‐1β and IL‐8 in GEC. IL‐1 inhibitors did not affect A. actinomycetemcomitans‐induced IL‐8, although they inhibited IL‐8 induced by IL‐1α or IL‐1β. In conclusion, GEC are a major source of IL‐1α and IL‐1β in the periodontium, which in turn induce additional inflammatory mediators such as IL‐8. Therefore GEC can be a potential target for therapeutic intervention in the future.
Effects of prolonged gum chewing on pain and fatigue in human jaw muscles
Authors: Farella Mauro , Bakke Merete , Michelotti Ambra , Martina Roberto
Published: March 2003

Gum chewing has been accepted as an adjunct to oral hygiene, as salivary stimulant and vehicle for various agents, as well as for jaw muscle training. The aim of this study was to investigate the effects of prolonged gum chewing on pain, fatigue and pressure tenderness of the masticatory muscles. Fifteen women without temporomandibular disorders (TMD) were requested to perform one of the following chewing tasks in three separate sessions: chewing a very hard gum, chewing a soft gum, and empty‐chewing with no bolus. Unilateral chewing of gum or empty chewing was performed for 40 min at a constant rate of 80 cycles/min. In each session, perceived muscle pain and masticatory fatigue were rated on visual analog scales (VAS) before, throughout, and after the chewing task. Pressure pain thresholds (PPTs) of masseter and anterior temporalis muscles were assessed before and immediately after the chewing tasks, and again after 24 h. The VAS scores for pain and fatigue significantly increased only during the hard gum chewing, and after 10 min of recovery VAS scores had decreased again, almost to their baseline values. No significant changes were found for PPTs either after hard or soft gum chewing. The findings indicate that the jaw muscles recover quickly from prolonged chewing activity in subjects without TMD.
Antero‐posterior activity changes in the superficial masseter muscle after exposure to experimental pain
Authors: Türp Jens C. , Schindler Hans J. , Pritsch Maria , Rong Qiguo
Published: March 2003

The aim of this randomized, controlled, double‐blind study was to examine how the activation pattern of the masseter muscle changes during natural function when experimental pain is induced in a discrete anterior area of the muscle. In 20 subjects, three bipolar surface electrodes and three intramuscular fine‐wire electrodes (antero‐posterior mapping) were simultaneously attached above and in the right masseter muscle to record the electromyographic (EMG) activity during unilateral chewing before and after infusion of a 0.9% isotonic and 5% hypertonic saline bolus in the anterior area of the muscle. The activity of the contralateral masseter muscle was registered by surface electrodes. In addition, the development of pain intensity was quantitatively measured with a numerical rating scale (NRS). While both saline concentrations caused pain, the hypertonic solution evoked stronger pain. The experiments also provided evidence of a significant although differential activity reduction of the ipsilateral masseter muscle in the antero‐posterior direction. The activity reduction decreased with increasing distance from the location of the infusion. The results support the idea that the strategy of differential activation protects the injured muscle while simultaneously maintaining optimal function.
Effect of irradiation type (LED or QTH) on photo‐activated composite shrinkage strain kinetics, temperature rise, and hardness
Authors: Hofmann Norbert , Hugo Burkard , Klaiber Bernd
Published: March 2003

This study compares commercially available light‐emitting diode (LED) lights with a quartz tungsten halogen (QTH) unit for photo‐activating resin‐based composites (RBC). Shrinkage strain kinetics and temperature within the RBC were measured simultaneously using the ‘deflecting disc technique’ and a thermocouple. Surface hardness (Knoop) at the bottom of 1.5‐mm thick RBC specimens was measured 24 h after irradiation to indicate degree of cure. Irradiation was performed for 40 s using either the continuous or the ramp‐curing mode of a QTH and a LED light (800 mW cm−2 and 320 mW cm−2, respectively) or the continuous mode of a lower intensity LED light (160 mW cm−2). For Herculite XRV and Filtek Z250 (both containing only camphoroquinone as a photo‐initiator) the QTH and the stronger LED light produced similar hardness, while in the case of Definite (containing an additional photo‐activator absorbing at lower wavelength) lower hardness was observed after LED irradiation. The temperature rise during polymerization and heating from radiation were lower with LED compared to QTH curing. The fastest increase of polymerization contraction was observed after QTH continuous irradiation, followed by the stronger and the weaker LED light in the continuous mode. Ramp curing decreased contraction speed even more. Shrinkage strain after 60 min was greater following QTH irradiation compared with both LED units (Herculite, Definite) or with the weaker LED light (Z250).
Relationship between cytokine concentration in serum, pre‐ovulatory follicular fluid and icsi outcome
Authors: Hammadeh M.E. , Hoffmeister H. , Rosenbaum P. , Schmidt W.
Published: March 2003

This study was undertaken to determine the concentrations of fibroblast growth factor (FGF), stem cell factor (SCF) and soluble intracellular adhesion molecule (sICAM) in serum and follicular fluid (FF) of patients who became pregnant (G.I) and those who did not (G.II) after controlled ovarian hyperstimulation (COH) for intracytoplasmic sperm injection (ICSI) therapy and to find out whether any relationship could exist between cytokine concentration in serum, FF on the day of oocytes retrieval and ICSI outcome.

Serum and FF was collected from 75 consecutive patients undergoing COH for ICSI therapy after pituitary down regulation with GnRHa. Three ampules (225 IU) of (hMG) were administered daily from day 3–7 of the cycle. 10.000 IU HCG was given i.m. Follicular aspiration was performed 35–36 hr after HCG injection. These cytokines were measured with ELISA assay kits. The cytokine levels in the serum of (G.I) were 8.5 + 10.5 pg/mL, 235.8 + 81.10 ng/mL and 597.7 + 139.9 pg/mL and the corresponding concentrations of (G.II) were (6.4 + 3.6, 230.6 + 66.5 and 569.6 + 91.4, respectively). There was no significant difference between the cytokines concentration in serum and FF in the groups. However, in both groups, the concentration of FGF and SCF in FF was significantly higher (P = 0.0001 and P = 0.05) than the value observed in serum. Whereas, the sICAM concentration was significantly higher of serum in comparison to FF value. The two groups did not differ significantly for the total number of gonadotropin ampoules administered, oestradiol concentration on the day of HCG injection, number of retrieved oocytes, and fertilization rate. In conclusion: Group I (the 25 women who did become pregnant) and group II (the 50 women who did not become pregnant) did not differ significantly for any of these cytokines concentrations. Besides, the ICSI outcome was not related to serum or follicular fluid concentrations of FGF, sICAM and SCF concentrations.
Therapy with Dendritic Cells (DCs) control fetal abortion rate in the CBA/J × DBA/2J mouse model
Authors: Blois Sandra M. , Olmos Sofia , Zenclussen Ana C. , Soto Catalina A. , Chuluyan Eduardo , Arias Karina , Gentile Maria T. , Arck Petra C. , Margni Ricardo A.
Published: March 2003

DBA/2J mated‐CBA/J female mice are prone to a high incidence of fetal abortions. This fetal wastage can be dramatically reduced by immunizing the female mice with BALB/c, but not DBA/2J spleen cells during early gestation, whereas underlying mechanisms remain to be elucidated. Recently, DCs have been described at the feto‐maternal interface in human uterus. The aim of this study was to analyse the role of DCs in the maintenance of pregnancy in the CBA/J × DBA/2J model. Bone marrow derived DCs were generated from virgin female CBA/J mice (6–8 weeks old) by using a modified version of Inaba's et al. technique, some of the DCs were pulsed with paternal DBA/2 J antigens lisate ex vivo. Immunization with DCs of the CBA/J females took place twice before matings. Four different experimental groups were included: (1) no treatment control, (2) mice injected with conditionated medium (GM‐CSF), (3) immunized with DCs and (4) immunized with Ag‐pulsed DCs, n = 5, respectively. Therapy with paternal antigen lisate pulsed DCs as well as the therapy with DCs alone significantly reduced the abortion rate from 23.8% in the controls (1 + 2) to 2.2% in Group 3 and 5% in Group 4. Our results indicate a protective role of DCs at the feto‐maternal interface during pregnancy. Putative pathways are the induction of an immunological response necessary in the prevention of abortion, i.e. a Th2 milieu or the production of asymmetric antibodies. These preliminary results may foster additional experiments on DCs in reproductive biology, which may soon be translated into clinically relevant health applications in a novel area of DCs research.
Increased Expression of Chemokine Receptor 5 (CCR5) in Blood and Decidual Cells in A Mouse Model for Preeclampsia (PE)
Authors: Zenclussen Ana C. , Fest Stefan , Joachim Ricarda A. , Hagen Evelin , Klapp Burghard F. , Arck Petra C.
Published: March 2003

A multicascade of leukocyte–endothelial cell interactions is involved in lymphocytes trafficking into tissues. Leukocytes adhesion to the endothelium is mediated by the interaction between endothelial chemokines and their receptors expressed on leukocytes. CCR5 expression has been intensively studied over the last years, since it is not only a trafficking marker, but also a reliable marker to identify Th1 cells. Further, we recently reported a PE mouse model, with increased blood pressure, proteinuria and pathological kidneys accompanied by increased peripheral and local Th1 cytokines levels compared to normal pregnant mice (NP). Here, we investigated whether the extravasation of Th1 lymphocytes into decidua was mediated by CCR5 during experimental PE. We evaluated (1) cytokines secretion and CCR5 expression by immune blood cells and (2) cytokines secretion as well as CCR5 levels in decidual lymphocytes using flow cytometry in PE and NP mice.

We observed enhanced production of IL‐12, IFN‐γ and TNF‐α by immune blood cells and augmented number of CCR5+ peripheral cells in PE compared to NP mice. We verified increased production of decidual IL‐12, IFN‐γ and TNF‐α in PE mice. The number of CCR5+ decidual cells was enhanced in PE compared to NP mice.

We propose that Th1 lymphocytes migration into decidua is enhanced due to up‐regulated percentages of CCR5+ cells in experimentally induced PE, contributing to the alteration of the physiological parameters observed in this model.
Cytokine Changes in Postmenopausal Women Treated with Estrogens: a Placebo‐controlled Study
Authors: BERG GÖRAN , EKERFELT CHRISTINA , HAMMAR MATS , LINDGREN RICHARD , MATTHIESEN LEIF , ERNERUDH JAN
Published: March 2003

PROBLEM: Hormone replacement therapy (HRT) is being increasingly used in postmenopausal women. Sex steroids are known to affect the immune system in several ways, although this is mainly based on clinical observations and experimental studies.

METHOD OF STUDY: We studied the in vivo effects of transdermal estrogens (50 μg 17 β‐Estradiol/24 hr) on cytokine production in postmenopausal women. A total of 17 women were randomized to either placebo (n=7) or active estrogen therapy (n=10) for 14 weeks, with addition of oral medoxyprogesterone acetate 10 mg daily during the last 2 weeks in both groups. Secretion of the cytokines IFN‐γ, IL‐4, IL‐10 and IL‐6 in blood mononuclear cells was determined, spontaneously and after stimulation with common vaccination antigens and mitogen, using the cell ELISA technique.

RESULTS: IL‐6 production after stimulation with purified protein derivate (PPD) decreased in the estrogen treated group (P < 0.01). Mitogen‐induced IL‐6 production was reduced in the estrogen treated group in contrast to an increase in the placebo group, leading to a significant difference (P < 0.01) between the groups after 12 weeks of treatment. This difference was eliminated after an addition of progestagens for 2 weeks. No significant changes were noted for IFN‐γ, IL‐4 or IL‐10 in relation to estrogen or placebo treatment.

CONCLUSIONS: In the present controlled study, the main in vivo effect of estrogens was a decrease in IL‐6 production, indicating a possible beneficial effect of estrogen therapy.
The Expressions of Intracellular Cytokines in the Lymphocytes of Preeclamptic Patients
Authors: Darmochwal‐Kolarz Dorota , Rolinski Jacek , Leszczynska‐Gorzelak Bozena , Oleszczuk Jan
Published: March 2003

Darmochwal‐Kolarz D, Rolinski J, Leszczynska‐Gorzelak B, Oleszczuk J. The expressions of intracellular cytokines in the lymphocytes of preeclamptic patients. AJRI 2002; 48:381–386 © Blackwell Munksgaard, 2002

PURPOSE: The objective of the study was to investigate the intracellular expressions of T helper 1 (Th1) and Th2 cytokines in peripheral blood T lymhocytes and natural killer (NK) cells of patients with preeclampsia and women with uncomplicated pregnancy.

METHOD OF STUDY: Blood samples were taken from 20 patients with preeclampsia and 16 healthy pregnant women. Mononuclear cells were isolated from peripheral blood and stimulated for 5 hr at 37°C and 5% CO2. Next, the cells were stained with antibodies against surface markers of T‐cell subsets and NK cells. After fixation and permeabilization processes, the cells were stained with antibodies against intracellular cytokines – interleukin‐2 (Il‐2) and interferon‐γ (IFN‐γ) as well as Il‐10 and Il‐4. The intracellular expressions of Th1 and Th2 cytokines were determined using the flow cytometric method. Statistical analysis was performed using Mann–Whitney U‐test.

RESULTS: We found that in patients with preeclampsia the expressions of Il‐2 were significantly higher when compared with women with uncomplicated pregnancy. Furthermore, in the group of patients with preeclampsia the expressions of Il‐2 were higher in T CD 8+ lymphocytes than in T CD4+ cells. The expressions of IFN‐γ did not differ in CD4+ cells and CD8+ cells in both studied groups but they were higher in NK cells in the study group. The expressions of Il‐10 were lower in lymphocytes of preeclamptic patients when compared with controls. The expressions of Il‐4 did not differ in both studied groups.

CONCLUSIONS: These results suggest that in patients with preeclampsia there is Th1/Th2 imbalance, with predominant Th1 immunity.
Increased Expression of Cyclooxygenase‐2 in Local Lesions of Endometriosis Patients
Authors: CHISHIMA FUMIHISA , HAYAKAWA SATOSHI , SUGITA KENJI , KINUKAWA NORIKO , ALEEMUZZAMAN SHEIKE , NEMOTO NORIMICHI , YAMAMOTO TATSUO , HONDA MITSUO
Published: March 2003

PROBLEM: Human endometrial glands contain the highest levels of cyclooxygenase (COX), although whether it is COX‐1 and/or COX‐2 has not been previously determined. Overexpression of COX‐2 may result in the pathogenesis of endometriosis.

METHOD OF STUDY: Tissue sections were obtained from 28 premenopausal women undergoing laparotomy or laparoscopic surgery for benign conditions. Endometrium, ectopic endometriosis tissue and peritoneum were obtained at the time of surgery. Informed consents were obtained from all the patients participating in this study. Immunohistochemistry was performed on consecutive sections of paraffin‐embedded tissue using anti‐COX‐2 antibody. Expressions of COX‐2 mRNA in endometrium, ectopic endometriosis tissue, and peritoneum were quanti‐tavely determined by competitive reverse transcription–polymerase chain reaction (RT–PCR).

RESULTS: In the uterus, COX‐2 was localized in the endometrial epithelium. Eutopic endometrial surface epithelium contains more COX‐2 than does glandular epithelium. We observed more frequent and denser COX‐2 staining in the ectopic endometriosis implants when compared with eutopic endometrium. Level of COX‐2 mRNA in endometriosis was increased up to five times that of eutopic endometria.

CONCLUSION: Hyper activation of COX‐2 with abnormal prostaglandin generation is considered to contribute to the pathophysiology of endometriosis and disease progression.