bluuebunny/crossref_metadata_2025_split huggingface dataset:
Telepathology: current status and future prospects in diagnostic histopathology
Authors: Cross S S , Dennis T , Start R D 
Published: March 2003

Telepathology: current status and future prospects in diagnostic histopathologyTelepathology is the process of diagnostic histopathology performed on digital images viewed on a display screen rather than by conventional glass slide light microscopy. The technology of telepathology has radically improved over the past 5 years so that it is no longer the limiting factor in the diagnostic process. This review looks at the resources needed for dynamic and static telepathology, including image quality, computers and software interfaces, means of transmission and human resources. It critically analyses 32 published trials of telepathology, including some large prospective studies, in all areas of diagnostic histopathology including intraoperative frozen sections, routine and referral cases. New developments, including internet solutions and virtual microscopy, are described and there is analysis of the economics of telepathology within health care systems. The review concludes that all the necessary technology for telepathology is available, there is strong published evidence for a diagnostic accuracy comparable with glass slide diagnosis, in many contexts there is a clear‐cut economic argument in favour of telepathology, and that the technique should now be integrated into mainstream diagnostic histopathology.
Morphological alterations in rat peritoneal mast cells by stem cell factor
Authors: Kim H. M. , Shin H. Y. , Lee E. H. 
Published: March 2003

Stem cell factor (SCF) stimulates mast cell adhesion and, because SCF is produced normally in tissues, it may be a major factor responsible for the adhesion of mast cells to connective tissue matrix. We found that the morphology of rat peritoneal mast cells (RPMC) altered after the addition of recombinant murine SCF (rmSCF) in vitro. The ability of rmSCF to enhance morphological alteration was dose dependent and completely abolished by anti‐c‐kit ACK2 monoclonal antibody. Exposure of RPMC to transforming growth factor‐β1, wortmannin, genistein, herbimycin A, staurosporine, indomethacin and cytochalasin D before the addition of rmSCF antagonized rmSCF‐induced morphological alteration. However, nordihydroguiaretic acid had no effect. Many RPMC appeared to respond also to nerve growth factor (NGF) but the total number of cells with altered morphology was much greater when the culture was stimulated by rmSCF than by NGF. We suggest that morphological alterations of mast cells by rmSCF is an important step for the participation in adhesion to tissue under resident physiological conditions.NDGA, nordihydroguiaretic acid NGF, nerve growth factor PI3‐kinase, posphatidylinositol 3‐kinase rmSCF, recombinant murine stem cell factor RPMC, rat peritoneal mast cells SCF, stem cell factor.
Rat complement factor I: molecular cloning, sequencing and expression in tissues and isolated cells
Authors: Schlaf G. , Rothermel E. , Oppermann M. , Schieferdecker H. L. , Jungermann K. , Götze O. 
Published: March 2003

SummaryFactor I (FI) is a regulatory serine protease of the complement system which cleaves three peptide bonds in the α‐chain of C3b and two bonds in the α‐chain of C4b thereby inactivating these proteins. The human protein and the recently characterized mouse factor I are heterodimers of about 88 000 MW which consist of a non‐catalytic heavy chain of 50 000 MW which is linked to a catalytic light chain of 38 000 MW by a disulphide bond. For the screening of a rat liver cDNA library we used a hybridization probe produced by polymerase chain reaction (PCR) using degenerated primers which corresponded to conserved parts of the human and the murine factor I nucleotide sequences. One of the identified sequences, which had a length of 2243 base pairs (bp), contained the complete coding region and the whole 3′ untranslated region. The length of the coding region in rat consisted of 1812 bp followed by a 3′ untranslated region of 207 bp including the polyadenylation signal and the beginning of the poly A tail. Comparison of the rat cDNA‐derived coding sequence revealed identities of 87% to the mouse and of 78% to the human FI nucleotide sequence. The translation product of rat FI mRNA was 604 amino acid residues (aa) in length with an identity of 85% to the mouse (603 aa) and 69% to the human protein (583 aa). The comparison of the molecular mass predicted by the primary structure and derived from rat FI isolated from rat serum as detected in immunoblot analyses suggested a glycosylation of more than 20% of the total mass of the FI protein. Expression studies using reverse transcription (RT)–PCR assays indicated that FI‐specific mRNA could neither be identified in B cells, nor in T cells, monocytes or granulocytes from rat and human peripheral blood nor in rat peritoneal macrophages. These data were in agreement with the results of RT–PCR obtained with several human lymphoma cell lines (Jurkat, MOLT‐4, HUT102, Wil 2‐NS, Ramos, Raji, U937) all of which were devoid of FI‐specific mRNA. In accord with our data from two rat hepatoma cell lines (FAO and H4IIE) and one from man (HepG2) only isolated rat hepatocytes (HC) but neither Kupffer cells (KC), hepatic stellate cells (HSC; Ito cells) nor sinusoidal endothelial cells (SEC) expressed FI‐specific mRNA. FI mRNA was also detected in human umbilical vein endothelial cells (HUVEC) and in the uterus and small intestine of the rat. Spleen and lymph nodes did not contain any detectable FI‐specific mRNA.
Interleukin‐12 modulates T‐cell responses to microfilariae but fails to abrogate interleukin‐5‐dependent immunity in a mouse model of onchocerciasis
Authors: Hogarth P. J. , Bianco A. E. 
Published: March 2003

SummaryInfection of mice with microfilariae of Onchocerca lienalis induces high levels of protective immunity to reinfection, which is dependent on interleukin (IL)‐5 but not IL‐4. Here, we have investigated the effect of exogenous IL‐12 administration during either the priming or effector phases of the immune response. When administered during priming, IL‐12 induced down‐regulation of parasite‐specific serum immunoglobulin (Ig)E and up‐regulation of IgG2a. Antigen‐specific IL‐4 responses were strongly suppressed, whilst blood eosinophil levels were partially reduced. When administered during a challenge infection, IL‐12 did not significantly influence the balance of antibody isotypes, but partially reduced eosinophil production. Antigen‐specific IL‐4 responses were again completely ablated. Unusually, interferon‐γ (IFN‐γ) responses were not significantly affected following IL‐12 administration, either during priming or after challenge infections. Moreover, despite a fall in antigen‐specific IL‐5 production, the expression of IL‐5‐dependent immunity, as determined by reduction in worm recoveries, was fully maintained. These data demonstrate that parasite‐induced IL‐4 can be abrogated without affecting protective immunity to Onchocerca microfilariae in mice. In view of the established role of IL‐4 in pathogenesis, this may have important implications for the development of immunoprophylaxis aimed at microfilariae and the alleviation of pathology in onchocerciasis.
Lysosomal cathepsin B plays an important role in antigen processing, while cathepsin D is involved in degradation of the invariant chain in ovalbumin‐immunized mice
Authors: Zhang T. , Maekawa Y. , Hanba J. , Dainichi T. , Nashed B. F. , Hisaeda H. , Sakai T , Asao T , Himeno K. , Good R. A. , Katunuma N. 
Published: March 2003

SummaryWe previously reported that CA074, a specific inhibitor of cathepsin B, modulates specific immune responses from the T helper 2 (Th2) type to Th1 type in BALB/c mice infected with Leishmania major. In the present study, we found that a similar type of immune deviation was also induced in mice immunized with ovalbumin (OVA). However, treatment of mice with pepstatin A, a specific cathepsin D inhibitor, suppressed the OVA‐specific proliferation of lymphocytes and blocked the development of both Th1 and Th2 cellular responses. These inhibitors did not appear to have any direct influence in vitro on functions of naive lymphocytes. OVA antigen (47 000 MW) was digested mainly into 40 000 MW protein in vitro by lysosomal proteases from naive BALB/c mice, and its digestion was markedly inhibited by the addition of CA074, but not by addition of pepstatin A, during incubation. However, pepstatin A strongly suppressed the degradation of the major histocompatibility complex class II‐associated invariant chain (Ii) molecule in vivo and in vitro. Thus, cathepsin B appears to process antigens directed to preferential activation of Th2 cells, while cathepsin D may be responsible for the degradation of Ii, the processing of which is essential in initiating the antigen‐specific activation of Th1 and Th2 CD4+ T cells. These lysosomal proteases may have different functions in regulating immune responses.
A monoclonal antibody to the rat Crry/p65 antigen, a complement regulatory membrane protein, stimulates adhesion and proliferation of thymocytes
Authors: Arsenović‐Ranin N. , Vučević D. , Okada N. , Dimitrijević M. , Čolić M. 
Published: March 2003

SummaryA murine monoclonal antibody (mAb), 3F10, was produced by fusion of spleen cells obtained from mice immunized with a rat cortical thymic epithelial cell line (R‐TNC.1) stimulated with interferon‐γ and P3X myeloma cells. 3F10 recognized an antigen expressed both on thymocytes and non‐lymphoid cells in the thymus. Flow cytometry showed that 3F10 stained more than 98% thymocytes and 90% R‐TNC.1 cells. Immunoprecipitation and Western blot studies demonstrated that 3F10 reacted with molecules of 55000 and 65000 MW from both thymocyte and R‐TNC.1 cell lysates. 3F10 recognized the same antigen on Chinese hamster ovary cells transfected with rat Crry as did 5I2 mAb, confirming the specificity of 3F10 mAb for the rat homologue of mouse Crry/p65, a membrane‐bound complement regulatory protein. 3F10 mAb induced homotypic aggregation of thymocytes and exhibited an additive effect on the aggregation evoked by phorbol myristate acetate. The aggregation was dependent on active cell metabolism, intact cytoskeleton, divalent cations and activation of protein phosphatases 1 and 2A (as assessed by use of okadaic acid). In contrast, H‐7, HA1004 and genistein partially inhibited, whereas staurosporine potentiated the aggregation of thymocytes triggered by 3F10. 3F10 mAb also stimulated binding of thymocytes to the R‐TNC.1 line. Both homotypic and heterotypic adhesive interactions are mediated by leucocyte function‐associated antigen‐1 (LFA‐1). In addition, 3F10 stimulated proliferation of thymocytes induced by suboptimal concentrations of concanavalin A. These data suggest that rat Crry/p65 might be involved in the regulation of both cell adhesion and activation of thymocytes. This is a novel, non‐complement‐dependent function of Crry/p65.
Presence of B220 within thymocytes and its expression on the cell surface during apoptosis
Authors: Oka S. , Mori N. , Matsuyama S. , Takamori Y. , Kubo K. 
Published: March 2003

SummaryB220 is the full‐length splicing isoform of a tyrosine phosphatase CD45 and is predominantly expressed as a transmembrane protein on B cells. Other splicing isoforms of CD45 are yielded by alternative splicing of exons 4, 5 and 6. Recently, the expression of B220 on peripheral T cells during activation‐induced cell death has been reported. To investigate whether B220 is implicated in apoptosis of immature T cells, we analysed (by flow cytometry using the anti‐B220 monoclonal antibody, RA3‐6B2) the expression of B220 on mouse thymocytes undergoing X‐irradiation‐ and dexamethasone (DEX)‐induced apoptosis. The expression of B220 on thymocytes positive for Thy‐1 was induced by X‐irradiation or DEX treatment and increased with length of incubation. The expression of B220 was pronounced on the apoptotic hypodiploid cells in the fraction showing lower forward scattering values. Reverse transcription–polymerase chain reaction detected mRNA containing exons 4, 5 and 6 of CD45 in normal thymocytes as well as those exposed to X‐rays or DEX. Surprisingly, cytoplasmic B220 antigens were detected in a considerable fraction of normal thymocytes. Moreover, the expression level of the 220 000‐MW protein in normal thymocytes was similar to that in the thymocytes undergoing apoptosis. During apoptosis, the expression level of B220 antigen was reduced in the cytoplasm but, conversely, up‐regulated on the surface of thymocytes. These results suggest that B220 is constitutively expressed as a cytoplasmic form within thymocytes and possibly translocated to the cell membrane during apoptosis.
Specificity and binding kinetics of murine lupus anti‐DNA monoclonal antibodies implicate different stimuli for their production
Authors: Eivazova E. R. , McDonnell J. M. , Sutton B. J. , Staines N. A. 
Published: March 2003

SummaryThe origin and relative biological importance of the many different DNA‐reactive antibodies that appear in systemic lupus erythematosus are not well understood. A detailed analysis of their fine specificity and binding characteristics with DNA is a necessary step in understanding their biology. We have examined here two monoclonal antibodies (mAb) IV‐228 and V‐88 that are, respectively, characteristic of antibodies, which bind exclusively to single‐stranded (ss) DNA and to both double‐stranded (ds) DNA and ssDNA. By surface plasmon resonance (SPR) on BIAcore, we characterized the kinetics of binding of each antibody to synthetic ss and ds oligonucleotides. Antibody V‐88 and IV‐228 showed different patterns of reactivity for both ss and ds oligonucleotides, characterized by distinctly different kinetic parameters. Analysis of their binding kinetics indicates the importance of base composition in defining DNA epitopes, and shows that some epitopes, such as that recognized by mAb V‐88, are expressed on dsDNA and ssDNA, whereas others, as recognized by IV‐228, are not. The base preferences of V‐88 for ds GC‐rich structures over AT‐rich, and of IV‐228 for ss T‐rich structures, also reveal distinct differences between these antibodies. We conclude that the different binding properties of the antibodies will relate to their biological activities. The base preferences of the antibodies suggest that they might be induced by different immunological stimuli, such as those that could be provided by the various DNA fragments and structures released during programmed cell death.
Natural antibody‐induced intracellular signalling and growth control in C3H 10T½ fibroblast variants
Authors: Wang H. , Chow D. A. 
Published: March 2003

SummaryCell‐surface binding by natural antibody (NAb) places it well for controlling cell function directly through signalling. Flow cytometry revealed an instability of syngeneic NAb binding to C3H 10T½ fibroblast variants at 37°, which could be partially reduced by H7, an inhibitor of the pivotal signalling serine/threonine kinase, protein kinase C (PKC). Cells coated with purified NAb at 4° followed by a rise in temperature to 37° showed an increase in membrane expression of introduced rat PKC‐β1 and endogenous PKC‐α, in the PKC‐β1‐overexpressing PKC‐4 and v‐H‐ras‐producing I3T2.1, respectively. Tyrosine phosphorylation of membrane‐associated 60 000 MW protein including the tyrosine kinase src was markedly reduced. In addition, both the precoated NAb and numerous membrane molecules ranging from 20 000 to 220 000 MW were released into the supernatant, including the receptor‐like protein tyrosine phosphatase α (RPTP‐α). Furthermore, purified NAb reduced the growth of I3T2.1 cells in culture assessed as a decrease in total cell numbers and an increase in the proportion of cells in the G0/G1 phase of the cell cycle. Together, these data argue that the interaction of NAb with cell surface structures initiated a series of intracellular signalling events leading to the release of membrane molecules and over time the suppression of cell proliferation. This process could provide a biological mechanism for direct NAb control of activated cells in both physiological and pathological conditions.
Role of β1 and β2 subunits of the interleukin‐12 receptor in determining T helper 1/T helper 2 responses in vivo in the rat
Authors: Gillespie K. M. , Szeto C.‐C. , Betin V. M. , Mathieson P. W. 
Published: March 2003

SummaryInterleukin‐12 (IL‐12) responsiveness, and hence capacity to mount a T helper type 1(Th1) immune response, may be regulated via differential expression of the IL‐12 receptor β2 subunit at least in vitro in human and murine cells. To test whether a similar phenomenon operates in vivo in the rat we cloned and sequenced partial cDNAs for rat IL‐12Rβ1 and IL‐12Rβ2 subunits and analysed expression of these genes in vivo in two rat strains with different Th1/Th2 bias. After treatment with mercuric chloride (HgCl2), Brown–Norway rats develop Th2‐biased autoimmunity whereas Lewis rats do not develop autoimmunity, instead becoming resistant to Th1‐biased diseases to which they are normally susceptible. We report close sequence homology between the segments of the rat IL‐12R genes sequenced and corresponding mouse genes (95·6% and 92% for IL‐12Rβ1 and IL‐12Rβ2, respectively). Both Brown–Norway and Lewis rats express both β1 and β2 subunits of IL‐12 receptor in vivo in spleen; Brown–Norway rats express the β2 subunit at a lower level than Lewis rats. After HgCl2 treatment, IL‐12Rβ1 expression was not altered but there was down‐regulation of IL‐12Rβ2 expression in both strains. We conclude that relative under‐expression of IL‐12Rβ2 by Brown–Norway rats contributes to their Th2 bias, and that down‐regulation of IL‐12Rβ2 after HgCl2 administration in Lewis rats underlies subsequent resistance to induction of Th1‐biased diseases.
Mast cells enhance contraction of three‐dimensional collagen lattices by fibroblasts by cell–cell interaction: role of stem cell factor/c‐kit
Authors: Yamamoto T. , Hartmann K. , Eckes B. , Krieg T. 
Published: March 2003

SummaryReorganization of the extracellular matrix is important in many biological and pathophysiological processes, including tissue remodelling, wound healing, or cancer metastasis. The ability of cultured fibroblasts to reorganize and contract three‐dimensional type I collagen gels is regarded as an in vitro model for this process. In tissue fibrosis, complex interactions among fibroblasts, inflammatory cells and the extracellular matrix are taking place. Mast cells have often been discussed to play a role in several fibrotic conditions including scleroderma, scar formation, or wound healing. In this study, we examined the effects of mast cells on contraction of collagen lattices. The results demonstrate that co‐culture of dermal fibroblasts with a human mast cell line (HMC‐1) significantly enhanced contraction of the three‐dimensional collagen lattices, whereas mast cells alone failed to contract the gel. Addition of culture supernatants of mast cells did not enhance the speed of gel contraction, indicating the importance of cell–cell contact. Morphological analysis showed that mast cells were incorporated into the lattices. Histological examination also demonstrated that within the lattices, mast cells were localized in close contact to, or attached to, fibroblasts. As fibroblasts and mast cells are known to attach via stem cell factor (SCF)/c‐kit interaction when co‐cultured in monolayers, we also examined the effect of antibodies against SCF and c‐kit in this system. Addition of both antibodies inhibited gel contraction up to 70%. In contrast, antibodies against interleukin‐4 (IL‐4) and IL‐4 receptor did not affect gel contraction. These results indicate that mast cells enhance fibroblast‐mediated contraction of collagen lattices via direct cell–cell contact, mediated in part by SCF/c‐kit interactions.
Interleukin‐6 regulates the phenotype of the immune response to a tuberculosis subunit vaccine
Authors: Leal Irene S. , Flórido Manuela , Andersen Peter , Appelberg Rui 
Published: March 2003

SummaryWe investigated the role of interleukin‐6 (IL‐6) in the development of the immune response to a subunit vaccine against tuberculosis consisting of the culture filtrate proteins of Mycobacterium tuberculosis emulsified in the adjuvant dimethyldioctadecylammonium bromide (DDA). C57Bl/6 mice immunized with this vaccine developed a strong T helper 1 (Th1) response characterized by an increased production of interferon‐γ (IFN‐γ) secreted by CD4+ T cells. Neutralization of IL‐6 during in vivo priming resulted in marked reduction in the ability of T cells to secrete IFN‐γ and IL‐2 and to proliferate. IL‐6 gene‐disrupted mice primed with the vaccine showed a decrease in the number of IFN‐γ‐producing cells and an increase in IL‐4‐secreting cells as compared to control mice. In contrast, neutralization of IL‐6 during a boost of the vaccine in previously primed mice did not affect the development of IFN‐γ‐producing cells but still increased the number of IL‐4‐producing cells. Our work shows that IL‐6 plays a major role in the priming but not in the later expression of a Th1 response to a tuberculosis vaccine.
Apoptosis and peripheral blood lymphocyte depletion in coeliac disease
Authors: Di Sabatino Antonio , D'Alò Simona , Millimaggi Danilo , Ciccocioppo Rachele , Parroni Raffaella , Sciarra Giuseppe , Cifone Maria Grazia , Corazza Gino Roberto 
Published: March 2003

SummaryIn coeliac disease (CD) immunological abnormalities are not confined to the small bowel and it has been suggested that changes in peripheral blood lymphocytes (PBL), such as lymphopenia and increased T‐cell activation, may predispose to malignant or autoimmune complications of this condition. In the light of the recent findings about the Fas–Fas ligand (FasL) system in regulating lymphocyte homeostasis, the aim of the present study was to investigate peripheral lymphocyte Fas‐mediated apoptosis in CD to establish whether the homeostatic role of apoptosis in peripheral T‐cell selection is maintained. Moreover, because a soluble form of Fas has been described to be functionally implicated in the Fas signalling system, suggesting a relationship between some disorders and soluble Fas function, we measured levels of soluble Fas in sera of coeliac patients and analysed the relationship between these levels and the proportions of apoptotic and Fas+ PBL to further explore the function of the Fas–FasL pathway in this condition. Finally, we evaluated whether the increased prevalence of anticardiolipin antibodies, recently described in CD, could be related to PBL apoptosis in this condition. We demonstrated an increased apoptosis and higher levels of Fas and FasL expression in PBL isolated from untreated coeliac patients when compared to treated coeliac patients and controls. In addition, low levels of soluble Fas and a significant positive correlation between anticardiolipin antibodies and PBL apoptosis were found in untreated CD. Then, our results showed an increased susceptibility of PBL to undergo Fas‐mediated apoptosis in active CD. This increased apoptosis could be responsible for both lymphopenia and immunogenic exposure of phospholipids with subsequent production of autoantibodies.