bluuebunny/crossref_metadata_2025 huggingface dataset:
Inhibition of cytokine secretion from human leukemic mast cells and basophils by H1‐ and H2‐receptor antagonists
Authors: Lippert U. , Möller A. , Welker P. , Artuc M. , Henz B. M.
Published: March 2003

Abstract: H1‐type antihistamines have recently been reported to inhibit cytokine secretion from human and murine mast cells and basophils. In order to confirm and expand these studies, we have compared several H1‐blockers and the H2‐blocker ranitidine for their effect on TNF‐α, IL‐3, 6, 8 and GM‐CSF release from human leukemic mast (HMC‐1) and basophilic (KU812) cells, compared to dexamethasone. Cells were stimulated for 24 h with phorbol myristate acetate (25 ng/ml) and calcium ionophore A 23187 (2.5×10−7 M) alone or with the drugs added at 10−4 to 10−15 M, and production of cytokines was measured by ELISA. All antihistamines caused a dose‐dependent inhibition of TNF‐α release from HMC‐1 cells, with maximal effects at 10−12 M for azelastine, 10−9 M for loratadine and cetirizine, and 10−8 M for ranitidine. The inhibitory potency of H1‐blockers on cytokines from HMC‐1 cells was TNF‐α >IL‐8≥IL‐6≥IL‐3, with no significant effects on GM‐CSF. In KU812 cells which failed to secrete TNF‐α and GM‐CSF, the sequence was IL‐6 >IL‐8 after preincubation. Dexamethasone inhibited all cytokines, but ranitidine only TNF‐α and IL‐3. Antihistamines had no effect on calcium flux in resting or stimulated cells. At the mRNA level, inhibition was only seen with KU812 cells and IL‐8 in the presence of azelastine at 10−10 M. These data show thus distinct inhibitory patterns for different antihistamines during cytokine production from human mast cells and basophils which may contribute to the anti‐inflammatory effects of these drugs during treatment of allergic diseases.
Alterations in mast cells showing tryptase and chymase activity in epithelializating and chronic wounds
Authors: Huttunen M. , Aalto M.‐L. , Harvima R. J. , Horsmanheimo M. , Harvima I. T.
Published: March 2003

Abstract: Mast cells can be found in contact with epidermis in certain circumstances; especially in chronic inflammatory skin diseases and chronic ulcers, but the significance of this association is obscure. In this study, the association of mast cells with wound healing was studied by counting mast cells in the wound edges at different stages after wounding the donor site skin for pinch‐grafting. Chronic venous leg ulcers were biopsed for comparison. Tryptase‐ and chymase‐positive mast cells were stained enzyme‐histochemically for active proteinases. Both the number of tryptase‐positive, i.e. total mast cells, and chymase‐positive mast cells decreased during wound healing, but only the change in chymase‐positive mast cells was statistically significant (P≤0.03) the maximal decrease being 63% on day 7. No mast cells could be found in the vicinity of epithelialization margin. In venous leg ulcers, significantly more mast cells were present in the perilesional skin near the epithelium margin than in the wound bed (P=0.03), and mast cells were also seen in close contact with the basement membrane. Immunoreactivity for IL‐4 and TNF‐α in mast cells was studied to see if either of these molecules was associated with wound healing. In normally healing wounds, only a minority of mast cells were immunoreactive for these cytokines and no change in positive mast cell numbers could be seen during wound healing. In chronic wounds, IL‐4 was absent in mast cells, and TNF‐α positive mast cells were present only in perilesional skin and in small numbers. These results show that mast cells; especially chymase‐positive; decrease in number and can not be found in the epithelialization zone in normal wound healing, whereas tryptase‐positive mast cells are associated with delayed wound healing and epithelialization in chronic wounds. Thus it seems, that mast cells attempt to control hyperproliferation of epidermis in chronic wounds.
Dendritic cell activation by danger and antigen‐specific T‐cell signalling
Authors: McLellan A. D. , Bröcker E.‐B. , Kämpgen E.
Published: March 2003

Abstract: Recent transplantation, animal and in vitro studies suggest a dependence of some immune reactions on tissue damage. Although many factors involved in enhancing immune responses through tissue damage have yet to be identified, recent data suggests that one of the targets of these cellular stress factors is the bone marrow derived dendritic cell ( DC). DC are potent initiators of primary immune responses and hold the key to immune reactions through their ability to sense changes in their local environment and respond appropriately to induce T‐cell immunity, or possibly tolerance. In the lymph node, DC are also influenced by antigen‐specific signalling from T cells, which may extend and amplify DC antigen presenting capabilities, especially for the stimulation of cytotoxic responses. It now appears that both tissue damage and antigen‐specific T‐cell derived signals act together on the DC to promote the appropriate immune reaction to antigen. Thus DC antigen presenting behaviour is not only dependent on the context of antigen encounter in the periphery, but also on the availability of antigen‐specific T cells and their T‐cell receptor specificities.
Circulating levels of soluble E‐selectin, ICAM‐1 and VCAM‐1 in bullous pemphigoid during low‐dose methotrexate therapy
Authors: Dahlman‐Ghozlan K. , Heilborn J. D. , Stephansson E.
Published: March 2003

Abstract: Soluble iso‐forms of cellular adhesion molecules (sCAMs) have been described and reported to be elevated in various inflammatory diseases. Elevated levels of sE‐selectin have recently been detected and found to correlate with the number of blisters in bullous pemphigoid (BP) during oral corticosteroid therapy. In this prospective study we analysed levels of sCAMs in 10 elderly BP patients during low‐dose oral pulse methotrexate monotherapy. We used standardised ELISA kits for soluble intercellular adhesion molecule‐1 (sICAM‐1), soluble vascular cell adhesion molecule‐1 (sVCAM‐1) and sE‐selectin on 65 sera from 10 patients and 19 controls. Results were correlated with clinical parameters. Before therapy, we found significant elevation of sE‐selectin (P=0.004) and sVCAM‐1 (P=0.002) but not of sICAM‐1. sE‐selectin levels decreased during the efficient therapy and correlated with the number of blisters. Our results further support the proposition that sE‐selectin might be a future clinical and predictive tool; but whether the elevation of sVCAM‐1 also might reflect the disease activity in BP needs more investigation. The findings also indicate that BP might be more a cellularly mediated disease where interactions of different adhesion molecules play a crucial role.
Cytokine expression patterns distinguish HIV associated skin diseases
Authors: Breuer‐McHam J. N. , Ledbetter L. S. , Sarris A. H. , Duvic M.
Published: March 2003

Abstract: AIDS is known to cause a shift of cytokines in the periphery. However, predominant cytokines in skin of patients with HIV‐associated skin diseases have not been clearly defined. We hypothesized that there are distinct cytokine profiles that distinguish among the different clinical manifestations of AIDS‐related skin diseases. To test this hypothesis, lesional and non‐lesional skin was biopsied from 53 HIV+ patients with Kaposi's sarcoma (KS), psoriasis, and pruritus due to eosinophilic folliculitis, and from HIV negative controls with psoriasis or KS prior to therapy. Immunohistochemistry was performed with antibodies to tumor necrosis factor (TNF)‐α, interleukin (IL)‐10, interferon (IFN)‐γ, and interferon‐inducible protein (IP)‐10. HIV positive individuals included 10 with psoriasis, 14 with pruritus, and 15 with Kaposi's sarcoma. HIV negative controls included 12 with psoriasis and two with KS. Semi‐quantitative analysis of cytokine staining was confirmed by optical density using a digital imaging system on four representative skin sections from each disease. Optical density analyses were conducted using ANOVA and t‐tests. We found that epidermis overlying HIV+ Kaposi's sarcoma was hyperproliferative and was highest in IP‐10, IFN‐γ, and IL‐10 (P=0.0001). HIV+ pruritus was significantly highest in TNF‐α (P=0.0001) staining. HIV+ psoriasis represented an intermediate state for all four cytokines. Normal skin adjacent to lesions showed the same relative patterns, with lower intensities. Skin diseases seen frequently in the setting of HIV and immunodeficiency have relatively distinct levels and patterns of cytokine expression that may reflect immune dysfunction, reactivity to HIV and to opportunistic infections.
Expression of involucrin in normal, hyperproliferative and neoplastic mouse keratinocytes
Authors: Li E. R. , Owens D. M. , Djian P. , Watt F. M.
Published: March 2003

Abstract: Involucrin is a protein precursor of the epidermal cornified envelope. Although expression of the human protein has been documented extensively, studies in the mouse have been hampered by a shortage of good antibodies. We describe the production of recombinant mouse involucrin and preparation of rabbit antisera to the protein that work well by immunohistochemistry and Western blotting. We confirm that in normal mouse epidermis the onset of involucrin expression is in the upper spinous layers and inner root sheath of the hair follicle. Involucrin was also detected in the differentiating epithelial cells of normal tongue, oesophagus and bladder. Involucrin was expressed in a subpopulation of mouse keratinocytes cultured in standard or low calcium medium and the proportion of involucrin‐positive cells increased during suspension‐induced terminal differentiation. Western blotting of keratinocytes from several inbred mouse strains revealed a remarkable heterogeneity in the electrophoretic mobility of involucrin, reflecting inter‐strain variation in the number of tandem repeats in the protein. In the hyperproliferative epidermis of healing wounds involucrin was expressed in most of the suprabasal layers. In epidermal papillomas and carcinomas involucrin expression correlated well with degree of histological differentiation. The sites of expression of the mouse protein were thus the same as those previously reported for human involucrin. With the development of the new antibodies we anticipate that involucrin will become as widely used a marker of keratinocyte differentiation in the mouse as it is in the human.
Doxepin affects acetylcholine induced cutaneous reactions in atopic eczema
Authors: Groene D. , Martus P. , Heyer G.
Published: March 2003

Abstract: Background: Atopic eczema (AE) is a chronic inflammatory skin disease with strong itching as the prominent symptom. The pathology of itch is still in discussion, but acetylcholine (ACH) seems to be a relevant pruritogenic mediator in AE. Since efficient benefit on pruritus and excoriations has been demonstrated with tricyclic agents, we investigated how the topical treatment with doxepin (5%, Boehringer Standard, Mannheim, Germany), a tricyclic compound with anticholinergic properties, may influence ACH induced itch and cutaneous sensations (erythema, wheal, axonreflex flare). Methods: Eleven patients with AE were included in this double blind study. For 3 days we applied doxepin cream to a defined area on the volar forearm and basic ointment to the other side 4 times daily. On day 4, ACH and sodium chloride were i.c. injected into the pretreated arms. Vasoreactions and cutaneous sensations were measured similar to studies described in previous publications from our group. Results: Doxepin treatment over 3 days reduced ACH provoked flare size more than 53% (P<0.005) and wheal size about 48% (P<0.005) whereas the maximal antipruritic effect was similiar to the basic therapy. The itch intensity, which is expressed as the mean AUC value, was rated at 6.12 arbitrary units after the neutral cream application and 5.9 arbitrary units after doxepin. Conclusions: The clinical and experimental effectiveness of doxepin as an antipruritic drug has been known for years. However, studies focusing on ACH as a pruritogenic mediator have not been performed. The duration of the doxepin application in our study seems to be appropriate since flare and wheal development were diminished. The reason why doxepin did not develop more antipruritic action compared to the vehicle cream may be due to the fact that the doxepin free cream already possessed an antipruritic action in this experimental study design. This is probably caused by rehydrating and moisturizing effects.
The feasibility of quantitative analysis of androgen metabolism by use of single dermal papillae from human hair follicles
Authors: Niiyama S. , Happle R. , Hoffmann R.
Published: March 2003

Abstract: Androgenetic alopecia (AGA) is a dihydrotestosterone‐mediated process, characterized by continuous miniaturization of androgen sensitive hair follicles (HF). Although increased 5α‐reductase (5aR) activity in affected HF is a key feature in the pathogenesis of AGA, only little is known about the in vivo expression of 5aR within AGA‐affected HF. Recent studies have shown that the dermal papilla (DP) is the predominant site of type 2 5aR expression within the human HF, but direct measurements of 5aR activity in intact DP of AGA‐affected HF have not been reported so far, mainly because of technical problems. Hence there is a need for a reliable and sensitive method of measuring 5aR activity in fresh tissues. As a novel approach, we used freshly isolated, intact DP and a highly sensitive HPLC‐radiomatic flow scintillation system to measure 5aR. In this way we were able to measure 5aR even in small DPs from miniaturized HF. Our results show that DP from the occipital scalp express ex vivo considerable amounts of 5aR activity, but the measurable enzyme activities of individual DP differ considerably. Therefore the use of only one or two DP is at present not a reliable tool to analyze 5aR activity ex vivo.
Mast cells as initiators of immunity and host defense
Authors: Henz B. M. , Maurer M. , Lippert U. , Worm M. , Babina M.
Published: March 2003

Abstract: Until recently, mast cells have been viewed primarily as harmful because of their key role as effector cells of allergic and potentially lethal anaphylactic reactions. Their contribution to human health appeared instead to be limited to the elimination of parasites. There is, however, growing evidence for additional beneficial functions of mast cells, particularly regarding the initiation of acquired immune reactions. Thus, mast cells can phagocytize diverse particles, take up antigens, and express a number of receptors, particularly MHC class I and II antigens, ICAM‐1 and ‐3, CD43, CD80, CD86 and CD40L which allow them to interact with T and B lymphocytes. They can also secrete numerous cytokines that induce and enhance recruitment and functions of lymphocytes. Finally, there is good evidence that mast cells present e.g. pollen and bacterial antigens, respond to bacterial superantigens, but fail to react to endogenously produced antigens or superantigens. Mast cells can also activate B cells directly to produce IgE, but this activity and the ability to produce IL‐4 or IL‐13 is restricted primarily to basophil leukocytes and mucosal mast cells. Finally, recent evidence attributes a pivotal role to the cells in natural immunity to bacteria. There is also emerging evidence that mast cells can downmodulate the immune response. While these data require further clarification, the basic ability of mast cells to initiate innate and acquired immune reactions can no longer be questioned.
The eczema area and severity index (EASI): assessment of reliability in atopic dermatitis
Authors: Hanifin J. M. , Thurston M. , Omoto M. , Cherill R. , Tofte S. J. , Graeber M. , Evaluator Group The Easi
Published: March 2003

Abstract: Objective– To test the reliability of the eczema area and severity index (EASI) scoring system by assessing inter‐ and intra‐observer consistency. Design: Training of evaluators, application, and assessment over 2 consecutive days. Setting– An academic center. Patients– Twenty adults and children with atopic dermatitis (AD); cohort 1 (10 patients ≥8 years) and cohort 2 (10 patients <8 years). Interventions– None. Main outcome measure– The EASI was used by 15 dermatologist evaluators to assess atopic dermatitis in cohort 1 and cohort 2 on 2 consecutive days. Inter‐ and intraobserver reliability were analyzed. Results– Overall intra‐evaluator reliability of the EASI was in the fair‐to‐good range. Inter‐evaluator reliability analyses indicated that the evaluators assessed the patients consistently across both study days. Conclusions– This study demonstrated that the EASI can be learned quickly and utilized reliably in the assessment of severity and extent of AD. There was consistency among the evaluators between consecutive days of evaluation. These results support the use of the EASI in clinical trials of therapeutic agents for AD.
Evidence for local control of gene expression in the epidermal differentiation complex
Authors: Elder James T. , Zhao Xinping
Published: March 2003

Abstract: The epidermal differentiation complex (EDC), located on chromosomal band 1q21, consists of at least 43 genes that are expressed during keratinocyte differentiation. Indicative of a role for chromatin structure in tissue specificity of EDC gene expression, we identified an inverse correlation between expression and DNA methylation for two EDC genes (S100A2 and S00A6) in human keratinocytes and fibroblasts. 5‐azacytidine (5AC) and sodium butyrate (NaB) are two agents known to promote ‘open’ chromatin structure. To explore the relationship between chromatin structure and keratinocyte differentiation, we treated normal human keratinocytes (NHK) with 5AC or NaB, or with protocols known to promote their terminal differentiation. We then measured the steady‐state mRNA levels for several S100 genes, small proline rich region‐1, ‐2, and ‐3, loricrin, and involucrin by Northern blotting. 5AC and NaB each markedly increased expression of SPRR1/2 and involucrin in NHK. In contrast, expression of S100A2 was reduced by both agents, and by induction of keratinocyte differentiation. Moreover, while the clustered EDC genes displayed a general tendency to be expressed in epithelial cells, they displayed different patterns of cell type‐specific expression. These results indicate that local, gene‐specific factors play an important role in the regulation of EDC gene expression in the keratinocyte lineage and during keratinocyte terminal differentiation.
Evidence that the keratinocyte colony number is genetically controlled
Authors: Popova Natalia V. , Tryson Kimberly A. , Wu Kai Q. , Morris Rebecca J.
Published: March 2003

Abstract: We tested five inbred strains and two outbred stocks of female mice in a quantitative assay for clonogenic keratinocytes from the cutaneous epithelium. We found three significantly different subsets of colony counts such that: C57BL/6 ≫ C3H = DBA/2 = SENCAR = BALB/c > FVB = CD−1 in culture conditions optimized for CD−1 0. C57BL/6 and BALB/c, two inbred parental strains, were chosen for further analysis. The F1 generation of these two parental strains had an intermediate number of colonies. The keratinocyte colony number from the two backcross generations was significantly different, while the colony number in the F2 generation was intermediate between the two backcrosses. We conclude that the number of keratinocyte colonies represents a new genetically definable quantitative trait. Analysis suggests that this trait is multigenic where the genes have an additive but not necessarily equal effect. We have therefore laid the foundation for identifying these stem cell regulatory genes, which may provide a new perspective on the mechanism of carcinogenesis and a new target for gene therapy.
Life‐history variation in perch (Perca fluviatilis L.) in five neighbouring Norwegian lakes
Authors: Heibo E. , Vøllestad L. A.
Published: March 2003

Abstract – We have studied the population dynamics and reproductive biology of perch (Perca fluviatilis L.) from five neighbouring lakes in Norway. The lakes vary in water quality, varying from strongly acidified (pH≤5.0) to not acidified (pH≥6.0). Estimated annual survival rates did not differ among populations. There was no strong relationship between age at maturity in female perch and acidification. However, age at maturity was lower for female perch from the most acidic lake (c. 2 years) compared with perch in the four other lakes (2.5–3.5 years). There was no relationship between acidification level and size‐adjusted egg size, fecundity or reproductive investment (Gonadosomatic index – GSI). Perch in the most acidified lake showed large variation in egg size, the lowest length‐specific fecundity, the highest GSI, and the lowest age at maturity. Environmental stress, such as acidification, induces a number of changes to a lake community. It is therefore difficult to predict how acidification will influence the population dynamics and reproductive biology of a perch population.
Distribution of non‐collagenous dentin matrix proteins and proteoglycans, and their relation to calcium accumulation in bisphosphonate‐affected rat incisors
Authors: Ohma Noriko , Takagi Yuzo , Takano Yoshiro
Published: March 2003

It has been reported that multiple injections of 1‐hydroxyethylidene‐1,1‐bisphosphonate (HEBP) to rats prevent mineralization of incisor dentin, thereby revealing high concentrations of calcium in the non‐mineralized matrix of circumpulpal dentin. To identify the molecules responsible for calcium accumulation in circumpulpal dentin matrix, rats were injected daily with HEBP (8 mg P/kg) for 7 d, and the incisors processed for various histochemical and immunohistochemical staining of non‐collagenous matrices of dentin. Cuprolinic blue reactions for proteoglycans (PGs) were equally distributed in non‐mineralized matrix of mantle and circumpulpal dentin layers. Dentin sialoprotein (DSP) and osteopontin (OPN) immunoreactions were found in non‐mineralized circumpulpal dentin matrix, but not in mantle dentin. In normal incisors, however, predentin matrix showing significant DSP immunoreactivity was negative for Ca‐GBHA reactions. HEBP‐affected, non‐mineralized OPN immunopositive bone matrix was also non‐reactive for calcium. From these observations, neither PGs, OPN nor DSP appear to be responsible for calcium accumulation in HEBP‐affected circumpulpal dentin. Stains‐all reactive component, possibly dentin phosphoprotein (DPP), only showed the same distribution as that of Ca‐GBHA in both HEBP‐affected and normal dentin matrix, implicating a possible contribution of DPP to calcium accumulation in circumpulpal dentin and, hence, to appositional mineralization of dentin.
Gender‐specific differences in temporomandibular retrodiscal tissues of the goat
Authors: Mariotti Angelo , Rumpf Deborah , Malakhova Olga , Cooper Brian
Published: March 2003

Healthy, adult, male and female goat temporomandibular retrodiscal tissues were characterized to determine if biochemical differences existed between the genders. RNA concentrations were not different between male and female retrodiscal tissues; however, the DNA concentration in female retrodiscal tissues was 82% greater than in male retrodiscal tissues. Collagen concentrations were significantly greater in male retrodiscal tissues, and this was reflected in significant gender differences of type I and III collagen concentrations. More specifically, male temporomandibular retrodiscal tissues contained 70% more type I collagen and 119% more type III collagen when compared to female retrodiscal tissues. These differences in collagens and DNA reflect a gender difference in temporomandibular retrodiscal tissue composition that underlies divergent biomechanical and neurophysiological properties.
Clinical wear performance of eight experimental dental composites over three years determined by two measuring methods
Authors: Söderholm Karl‐Johan M. , Lambrechts Paul , Sarrett David , Abe Yasuhiko , Yang Mark C.K. , Labella Roberto , Yildiz Esra , Willems Guy
Published: March 2003

The effect of matrix selection, filler composition, filler silanization, operator variations, and test site (dental clinic) on the wear rate of eight composites were evaluated. The wear was measured on replicas using both a microscopic and a laser scanning measuring method. The average wear rate on contact‐free surfaces was 9.2±4.2 µm/month with the microscopic measurement and 8.5±3.7 µm/month with the laser scanner over the 36‐month period. The urethane‐based composites performed significantly better than those which were bisGMA‐based. Restorations placed at one dental clinic showed significantly lower initial wear. There was also a significant difference between the operators that was most pronounced during the first 6 months. The other variable (filler composition and silane treatment) did not affect the wear rate significantly.
An adenoviral system for tetracycline‐regulated TGF‐β expression mediates a reversible cell cycle arrest
Authors: Wettergreen Matthew A. , Hunniford John W. , Crawford John M. , Adami Guy R.
Published: March 2003

The ability to regulate the proliferation of normal cells in a reversible manner would be a useful adjunct to some clinical therapies, including many types of cancer chemotherapy and surgery for periodontal regeneration. While the application of recombinant growth factors and cytokines to target cells is a logical approach to regulate cell proliferation, the high turnover rates of these peptide factors often make this approach impractical. Recombinant adenoviral vectors can be used to direct the expression of transgene products such as growth factors in many cell types in vitro and in vivo. We have adapted the tetracycline‐regulated expression system to allow regulated transforming growth factor (TGF)‐β1 expression using recombinant adenovirus. We demonstrate that infection with a recombinant TGF‐β‐encoding virus system in primary human oral keratinocytes and in a lung epithelial cell line is sufficient to allow a cell cycle arrest that is reversible upon tetracycline addition. This inhibition is efficient even after the infection of a minority of cells in a population. These results highlight the possibility of using low level infection with recombinant adenovirus to cause short‐term blocks on cell proliferation.
Late effects of radiotherapy on oral mucosa in humans
Authors: Handschel Jörg , Sunderkötter Cord , Kruse‐Lösler Birgit , Prott Franz‐Josef , Meyer Ulrich , Piffko Josef , Joos Ulrich
Published: March 2003

In order to gain further understanding of the late effects of radiotherapy on oral mucosa, we analysed the histomorphological alterations, the cell populations in the subepithelial tissue, and the endothelial expression pattern of different adhesion molecules. Biopsies were taken from patients before irradiation, directly after 60 Gy, and 6–12 months after radiotherapy. Besides the histomorphological evaluation of the vessels, the endothelial expression of ICAM‐1, VCAM‐1 and E‐selectin was determined as well as the distribution of LFA‐1‐, Mac‐1‐, VLA‐4‐, RM3/1‐, 27E10‐ and 25F9‐bearing cells in the subepithelial tissue. The expression of ICAM‐1 was downregulated after radiotherapy, whereas the percentage of LFA‐1‐ and VLA‐4‐bearing cells increased. VCAM‐1 remained at low levels. The subepithelial infiltration was still dominated by RM3/1‐positive macrophages. The number of vessels decreased, while the lumen of the remaining vessels increased. In conclusion, the late effects of radiotherapy are characterized by a decreased number of blood vessels and by significantly different expression patterns of the adhesion molecules studied, and of integrins and macrophage subpopulations, compared to the conditions before irradiation and directly after irradiation with 60 Gy.
Effects of radiation on Ca2+ signaling in salivary epithelial cell lines transfected with Bcl‐2 and Bcl‐XL
Authors: Sun Xiuhua , Liu Xiao‐bing , Martinez J. Ricardo , Dang Howard , Zhang Guo H.
Published: March 2003

The effects of radiation on the Ca2+ signaling system in HSY cells transfected with the Bcl‐2 or Bcl‐XL gene were studied. Bcl‐2 overexpression did not alter carbachol (CCh)‐elicited initial increase in cytosolic free Ca2+ concentrations ([Ca2+]i), but Bcl‐XL overexpression dramatically reduced this response. Exposure to 10 Gy γ‐ray did not alter basal [Ca2+]i. By contrast, the CCh‐stimulated initial [Ca2+]i increase was reduced at 0.5 and 4 h post‐irradiation in all cell types and remained decreased at 24 h in wild‐type and control‐transfected cells, but recovered in Bcl‐2‐ and Bcl‐XL‐transfectants. The formation of inositol 1,4,5‐trisphosphate (IP3) in response to CCh at 4‐h post‐irradiation was decreased in wild‐type and control‐transfected cells, but not in Bcl‐2 and Bcl‐XL transfectants. The capacity of the IP3‐sensitive Ca2+ store was significantly reduced by radiation in all cells except Bcl‐XL transfectants. Ca2+ influx after stimulation with CCh was suppressed by exposure to radiation in wild‐type and control‐transfected cells, but not in Bcl‐2‐ and Bcl‐XL‐transfectants. However, radiation enhanced Ca2+ influx activated by thapsigargin in all cell types. These results suggest that 1) radiation diminishes IP3 formation and Ca2+release in response to CCh, but potentiates the store‐operated Ca2+ influx; and 2) overexpression of Bcl‐2 or Bcl‐XL partially protects cells from radiation‐induced inhibition of Ca2+ signaling.
Monoclonal antibody against Porphyromonas gingivalis hemagglutinin inhibits hemolytic activity
Authors: Hosogi Yumiko , Hayakawa Mitsuo , Abiko Yoshimitsu
Published: March 2003

Porphyromonas gingivalis has been implicated as an important pathogen in the development of adult periodontitis. This bacterium possesses hemagglutinating and hemolytic activities to attach and lyse erythrocytes. Hemolysis by this oral pathogen functions to provide heme‐containing molecules for growth in the periodontal pocket. We previously constructed a monoclonal antibody using P. gingivalis vesicles as the immunogen, designated as MAb‐Pg‐vc, which inhibited vesicle‐associated hemagglutinating activity. Furthermore, we cloned the gene encoding 130‐kDa hemagglutinin (130‐kDa HAG) and identified its functional motif for attachment to erythrocytes. Generally, bacterial cell attachment to erythrocytes is an important initial step for expressing hemolysis activity. In the present study, we examined the effect of MAb‐Pg‐vc on the hemolytic activity of P. gingivalis cells. The MAb‐Pg‐vc significantly inhibited the hemolytic activity and, further, this inhibitory activity was reduced by the synthetic peptide of the 130‐kDa HAG functional motif.
Inhibition of HIV‐1 IIIB and clinical isolates by human parotid, submandibular, sublingual and palatine saliva
Authors: Bolscher Jan G. M. , Nazmi Kamran , Ran Leonie J. , Van Engelenburg Frank A. C. , Schuitemaker Hanneke , Veerman Enno C. I. , Amerongen Arie V. Nieuw
Published: March 2003

Human saliva is known to possess components that decrease the HIV‐1 infectivity in vitro. The mechanism of how these components inhibit the infectivity is still not clear on the molecular level. The purpose of this study was to discriminate between serous and mucous components with respect to inhibitory capacity and site of action. We have used total saliva and saliva from the major (sero)mucous glands: submandibular gland, sublingual glands, and glands in the palate, in comparison with the serous parotid glands. HIV‐1 IIIB and primary variants were incubated with saliva, and inhibition of HIV‐1‐infection was determined by analysing the cytopathic effect on MT‐2 cells. Mucous saliva, as well as serous saliva, contained high molecular weight components that reduced HIV‐1‐infectivity, at least partially by entrapment of the virus particles. Lower molecular weight components in all types of saliva possessed strong HIV‐1 neutralizing capacity. Using pro‐viral DNA synthesis by reverse transcription as a discrimination point in the replication cycle, the results indicated that part of the saliva samples acted before, but others after, this point. In conclusion, saliva inhibits HIV‐1‐infection by the action of high molecular weight components in combination with low molecular weight components from serous as well as mucous saliva, affecting different stages of the infection cycle.
Hardness and in vitro wear of a novel ceramic restorative cement
Authors: Sunnegårdh‐Grönberg Karin , Peutzfeldt Anne , Van Dijken Jan W. V.
Published: March 2003

The aim of the present work was to compare a new ceramic restorative cement for posterior restorations, DoxaDent, with other types of tooth‐colored materials for direct use as regards hardness and in vitro wear. Four hybrid resin composites, one polyacid‐modified resin composite, one resin‐modified glass ionomer cement, one conventional glass ionomer cement, one zinc phosphate cement, an experimental version as well as the marketed version of the ceramic restorative cement, were investigated. Hardness of the materials was tested with the Wallace indentation tester and wear was tested with the ACTA wear machine. All tests were carried out on 2‐wk‐old specimens. DoxaDent was as hard as the zinc phosphate cement and the hardest resin composite. The ceramic restorative cement wore significantly more than the resin composites, the same as the zinc phosphate cement, and less than the glass ionomer cements. No correlation between hardness and wear was found. It can be concluded that the ceramic restorative cement is a rather hard material but with a relatively low wear resistance.
Effects of various antiplaque agents on fructosyltransferase activity in solution and immobilized onto hydroxyapatite
Authors: Steinberg Doron , Bachrach Gilad , Gedalia Itzhak , Abu‐Ata Samir , Rozen Ramona
Published: March 2003

Fructosyltransferases (FTFs) are extracellular enzymes which synthesize fructans from sucrose. Cell free FTFs are found in the dental plaque biofilm as well as in saliva. Fructans play an important role in the progression of dental caries, mainly by serving as an extracellular nutrition reservoir for bacteria. The objective of the present study was to compare the effects of several antiplaque agents on the synthesis of fructans by FTF immobilized on hydroxyapatite (HA) or in solution. The effect of chlorhexidine, cetylpyridinium chloride, sodium lauryl sulfate and Tween on FTF activity was tested using radioactive assays. Their effect on fructan structure was tested using circular dichroism–optical rotative dispersion (CD‐ORD) analysis and Fourier transform infrared (FT‐IR) spectroscopy. Our results show that the antiplaque agents tested had an inhibitory effect on FTF activity both in the immobilized phase and in solution, although the inhibitory effect was more pronounced in solution. Structural changes in fructans, due to the presence of the antiplaque agents, were recorded as additional C–H or O–H bands demonstrated in FT‐IR analysis. However, non‐significant changes in peak location were detected in CD‐ORD spectrum between fructans synthesized in solution and on HA surfaces, and after treatment with the different antiplaque agents. Our study shows that several antiplaque agents may affect FTF activity and the synthesis of fructans by FTF, immobilized on hydroxyapatite or in solution.
In vitro study of caries detection through sound dentin using a laser fluorescence device, DIAGNOdent
Authors: Iwami Yukiteru , Shimizu Ayako , Yamamoto Hiroko , Hayashi Mikako , Takeshige Fumio , Ebisu Shigeyuki
Published: March 2003

The objective of this study was to investigate the feasibility of proximal surface caries detection using a laser fluorescence device, DIAGNOdent, through the sound dentin surrounding the cavity wall after removal of occlusal caries. Extracted sound human molars were ground to prepare horizontal and vertical dentin plates 1.4 mm thick. Extracted carious human molars were reduced in thickness horizontally from the occlusal surfaces until exposure of the dentin caries. The dentin plate was placed on the carious tooth. Before and after gradual thinning of the dentin plate from approximately 1.4 mm to 0.2 mm thick, the dentin caries was measured 10 times through the plate by the laser fluorescence device with a cone‐shaped tip or a broad tip. When the dentin plate was reduced to less than 0.3 mm thick (using a combination of a horizontal plate and cone‐shaped tip) or 0.2 mm thick (using the other combinations), the values measured with the laser fluorescence device were significantly larger than the values before reducing the thickness of the dentin. Based on these results, the device offers potential use as one of the screening tests for proximal surface caries detection through sound dentin when the sound dentin between tip and caries is thin.
Immunoregulatory properties of decidua macrophages
Authors: Sedlmayr Peter , Cupurdija Kristijan , Azzola Dagmar , Dohr Gottfried , Wintersteiger Reinhold , Morales Pedro , Petroff Margaret G. , Hunt Joan S. , Gratchev Alexei , Goerdt Sergij
Published: March 2003

About 15–20% of bone marrow derived cells in the endometrium and the decidua are CD14‐positive macrophages coexpressing the scavenger receptor CD163. The costimulatory molecule CD86 (B7‐2) but not CD80 (B7‐1) is found expressed on first trimester and term decidua macrophages (D‐Mph). The number of D‐Mph increases at the implantation site in the basal decidua, where they are in contact with the HLA‐G‐positive invading extravillous cytotrophoblast. As D‐Mph express the inhibitory receptors CD85d (ILT4, LIR‐2) and CD85e (ILT2, LIR‐1), they are potentially susceptible to inhibition by class Ia and class Ib molecules. D‐Mph may have specific functions relating to successful pregnancy, and phenotypic and functional properties may differ between early and term pregnancy. A number of data make their molecular repertoire comparable to those of alternatively activated macrophages which are immunosuppressive. For instance, first trimester decidua macrophages express IL‐10 and the MS‐1 high molecular weight antigen on the cell surface fibronectin and F XIIIa are detectable. Functional data are required to complement these studies.
An Optimized Technique for Separation of Human Decidual Leukocytes for Cellular and Molecular Analyses
Authors: NAGAEVA OLGA , BONDESTAM KARIN , OLOFSSON JAN , DAMBER MATS‐GÖRAN , MINCHEVA‐NILSSON LUCIA
Published: March 2003

PROBLEM: Human decidua contains a significant number of leukocytes, phenotypically and functionally different from peripheral blood. In vitro studies of decidual leukocytes require constant improvement of cell isolation technique in order to obtain highly purified viable and functionally active cells.

METHOD OF STUDY: Optimized isolation procedure of decidual leukocytes, based on non‐enzymatic cell dispersion, Percoll gradient centrifugation and purification step involving immunomagnetic beads is compared with a similar procedure with Lymphoprep gradient.

RESULTS: The method based on Percoll gradient gave a viable leukocyte fraction with higher amount of αβT‐ and γδΤ cells compared with Lymphoprep gradient. The leukocytes exhibited a well‐preserved ultrastructure and surface marker expression and were suitable for molecular analysis. The magnitude and the kinetics of their proliferative response were comparable with peripheral blood lymphocytes.

CONCLUSIONS: The results provide an optimized method for isolation and enrichment of decidual leukocytes, suitable for phenotypic, ultrastructural, molecular and functional analyses.
Interleukin 2 Receptor Gamma Chain (γc) Knockout Mice Show Less Regularity in Estrous Cycle but Achieve Normal Pregnancy Without Fetal Compromise
Authors: MIYAZAKI SATOMI , TANEBE KYOKO , SAKAI MASATOSHI , MICHIMATA TOSHIHIKO , TSUDA HIROSHI , FUJIMURA MASAKI , NAKAMURA MASATAKA , KISO YASUO , SAITO SHIGERU
Published: March 2003

PROBLEM: Implication of cytokines in pregnancy suggested but remains to be established. We studied the effect of cytokines on reproductive functions such as ovulation and pregnancy, with mutant mice lacking interleukin 2 receptor γ (IL‐2Rγ), the so‐called common γ chain (γc), which is shared among receptors for multiple cytokines such as IL‐2, IL‐4, IL‐7, IL‐9 and IL‐15.

METHOD OF STUDY: Regularities of estrous cycles were observed by vaginal smear. Ovaries stimulated with postmenopausal serum gonadotropin (PMSG) were examined for the ovarian capacities. The uteri at 13 days of gestation were used for histological analysis of the maternal–fetal interface.

RESULTS: The estrous cycles in γc knockout (γc KO) mice were irregular compared with wild‐type mice, although, the mutants could become pregnant. No uterine natural killer (uNK) cell was found in the uterus at 13 days of pregnancy, and poor decidual formation and thickness of blood vessel walls were observed. Apparent differences were not seen in the numbers and weight of the fetuses between wild type and mutant animals, and fetuses were not compromised throughout pregnancy.

CONCLUSIONS: The γc KO mice showed irregular estrous cycle but they could carry out normal pregnancy despite lacking uNK and cytokines actions of IL‐2, 4, 7, 9 and 15.
Immunization for Protection of the Reproductive Tract: A Review
Authors: RUSSELL MICHAEL W.
Published: March 2003

PROBLEM: Local application of non‐replicating antigens to the female reproductive tract is ineffective in stimulating the common mucosal immune system, and induces only weak genital antibody responses. Studies of immune responses to genital infections such as gonorrhea also support the concept that, lacking mucosal immune inductive sites, the reproductive tract is ill‐equipped to mount effective immune responses.

METHOD OF STUDY: Intranasal (i.n.) and intravaginal (i.vag.) routes of immunization of mice with a protein antigen coupled to cholera toxin (CT) B subunit, or genetically engineered as chimeric proteins with the A2/B sunbunits of CT or type II heat‐labile enterotoxin, were compared for their ability to induce specific antibody responses in vaginal fluids, saliva, and serum.

RESULTS: Mice immunized i.n. developed substantially stronger vaginal immunoglobulin A (IgA) and immunoglobulin G (IgG) and serum IgG and IgA antibodies, than those immunized i.vag. which also failed to develop salivary antibodies. Vaginal antibody responses induced i.n. persisted for at least 1 year, and were recallable by booster immunization after a prolonged period.

CONCLUSIONS: Such alternative strategies for inducing potent genital antibody responses offer the prospect of prophylactic immunization against genital infections. Further studies are required to evaluate their applicability to humans, and to comprehend the cellular and molecular mechanisms involved in delivering effective immune responses to the reproductive tracts.
Accumulation of CD16+ Cells with Secretion of Ksp37 in Decidua at the End of Pregnancy
Authors: HAYANO CHIKAKO , KOI HIDEKI , OGAWA KAZUYUKI , NAGATA KINYA , MATSUMOTO YUKIE , NAKAMURA MASATAKA , ASO TAKESHI
Published: March 2003

PROBLEM: Maternal cellular immunity is thought to be in a state of tolerance during pregnancy, but the precise mechanism of immunomodulation is not yet known. We investigated a novel serum protein, killer‐specific secretory protein of 37 kDa (Ksp37), produced by cytotoxic lymphocytes, during pregnancy.

METHOD OF STUDY: The level of Ksp37 was determined by enzyme linked immunosorbent assay (ELISA) in the sera of healthy pregnant women. Intracellular Ksp37 expression in mononuclear cells, isolated from peripheral blood and decidua at parturition, was examined with a flow cytometer.

RESULTS: Serum Ksp37 levels significantly increased at late pregnancy, compared with non‐pregnant controls and the first trimester of pregnancy. The flow cytometric analysis exhibited that Ksp37 was mainly expressed in CD16+ natural killer (NK) cells in decidua of term placenta.

CONCLUSIONS: Serum Ksp37 level was elevated at late gestational period. CD16+ NK cells in decidua seem to be a main maternal source of Ksp37. Innate immunity, with CD16+ NK cells, may play important roles near parturition.
Total Cortisol Levels are Reduced in the Periovulatory Follicle of Infertile Women with Minimal–Mild Endometriosis
Authors: SMITH M.P. , KEAY S.D. , MARGO F.C. , HARLOW C.R. , WOOD P.J. , CAHILL D.J. , HULL M.G.R.
Published: March 2003

PROBLEM: To measure and compare concentrations of total and free glucocorticoids with oocyte fertilizing capacity in the follicular fluid (FF) of women with minimal–mild endometriosis and tubal damage.

METHOD OF STUDY: Follicular fluid was collected from individual periovulatory follicles during oocyte retrieval for in vitro fertilization (IVF) in natural cycles. Total and free levels of cortisol and cortisone were measured using specific radioimmunoassays after chloroform extraction.

RESULTS: Cortisol concentrations in women with minimal–mild endometriosis were significantly lower compared with controls (women with tubal infective damage) (258 versus 328 nmol/L, P < 0.02). There was no correlation between total or free concentrations of cortisol or cortisone and the fertilization capacity of the oocyte.

CONCLUSIONS: Total cortisol levels are lower in the follicles of women with endometriosis. Our findings provide further evidence of follicular dysfunction contributing to the subfertility associated with minimal–mild endometriosis.
Distinct T Cell Subsets and Cytokine Production in Cultures Derived from transformation Zone and Squamous Intraepithelial Lesion Biopsies of the Uterine Cervix
Authors: Jacobs Nathalie , Renard Isabelle , Al‐Saleh Walid , Hubert Pascale , Doyen Jean , Kedzia Witold , Boniver Jacques , Delvenne Philippe
Published: March 2003

Jacobs N, Renard I, Al‐Saleh W, Hubert P, Doyen J, Kedzia W, Boniver J, Delvenne P. Distinct T cell subsets and Cytokine production in cultures derived from transformation zone and squamous intraepithelial lesion biopsies of the uterine cervix. AJRI 2003; 49:6–13 © Blackwell Munksgaard, 2003

PROBLEM: The characterization of lymphocytes issued from squamous intraepithelial lesions (SIL) and from the transformation zone (TZ), where the majority of SIL occur, is important to understand the role of immunity in SIL development.

METHOD OF STUDY: We compared lymphocyte populations of the TZ and SIL with those of normal exocervix, using a technique allowing for the isolation of lymphocytes, either from the epithelium or from the underlying stroma of small biopsies.

RESULTS: The majority of cells derived from the epithelium of all biopsies were CD8+ T cells. Some SIL‐derived cultures were characterized by an increased proportion of activated TCRγδ+. The production of the immunosuppressive cytokine IL10 was significantly higher in lymphocyte cultures from the normal TZ in comparison with the exocervix. A decreased percentage of effector T cells was observed in cultures derived from the stroma of normal TZ (TCRγδ+) or SIL (CD8+) in comparison with the exocervix.

CONCLUSIONS: Our results suggest that a low proportion of effector T cells and IL10 production could contribute to the predisposition of the TZ to the development of SIL and to the progression of SIL to cervical cancer.