bluuebunny/crossref_metadata_2025 huggingface dataset:
Laparoscopy‐assisted myomectomy of large uterine myomas
Authors: Tulandi T. , Youseff H.
Published: November 2003

Objectives To compare laparoscopic‐assisted myomectomy with myomectomy by laparotomy. Design Non‐prospective study. Patients Women with large uterine myoma of >16 weeks. Interventions Laparoscopic‐assisted myomectomy (n = 31) and myomectomy by laparotomy (n = 29). Results The mean diameter and the weight of the myoma were 12.7 ± 3.5 cm and 267.5 ± 36.0 g in the laparoscopy‐assisted myomectomy group and 9.1 ± 0.6 cm and 332.6 ± 62.9 g in the laparotomy group. The mean length of hospital stay was significantly shorter in women who underwent laparoscopic‐assisted myomectomy (3.2 ± 0.2 days) than in those who underwent myomectomy by laparotomy (5.1 ± 0.3 days). Conclusions Laparoscopic‐assisted myomectomy is as effective as myomectomy by laparotomy, but it is associated with a smaller abdominal incision and shorter hospital stay.
Laparoscopically assisted vaginal hysterectomy: a report of the first 300 completed procedures
Authors: Bolger B. S. , Lopes A. , Monaghan J. M.
Published: November 2003

Objectives To document the clinical, surgical and postoperative details collected prospectively for the first 300 laparoscopically assisted vaginal hysterectomies performed in the authors' unit. Design Prospective descriptive study. Setting Regional gynaecological oncology unit and two private hospitals. Subjects A total of 317 women scheduled for laparoscopically assisted vaginal hysterectomy (LAVH). Results In 94.6% of cases hysterectomy was achieved by LAVH. Significant complications occurred in 12.3% of scheduled cases. This included three patients requiring laparotomy following a successful LAVH; 11 patients requiring transfusion; eight patients with significant lateral laparoscopic port injury, and four patients with bladder injury. LAVH does not require greatly increased surgical time (median 60 min). The series identified a tendency towards the use of LAVH only for those patients where oöphorectomy was to be performed. Conclusions These data provide valuable information to help establish and interpret the results of future comparative studies.
Islet cell tumour with vacuolated lipid‐rich cytoplasm: a new histological variant of islet cell tumour
Authors: ORDÓÑEZ N.G. , SILVA E.G.
Published: November 2003

Aims: A previously unreported clear cell variant of islet cell tumour of the pancreas is described. Methods and results: By light microscopy, the cytoplasm of the clear cells was finely reticulated and vacuolated. Immunohistochemical and electron microscopy studies confirmed the endocrine nature of the tumour. Conclusions: The clear cell morphology was due to the accumulation of lipid in the cytoplasm.
Angiotropic lymphoma with endocrine involvement
Authors: SHANKS J.H. , HARRIS M. , HOWAT A.J. , FREEMONT A.J.
Published: November 2003

Aims: Two cases of angiotropic high‐grade B‐cell lymphoma are described in which involvement of endocrine glands was a prominent feature. Methods and results: One patient had marked unilateral adrenomegaly due to lymphoma within adrenal blood vessels. He is alive 5 years after presentation following combination chemotherapy. The diagnosis was made at autopsy in the second patient who presented with pyrexia of unknown origin and had hypothyroidism and a thyroid nodule. The thyroid gland and pituitary showed striking involvement by angiotropic lymphoma and the thyroid nodule was a cavernous haemangioma heavily colonized by lymphoma cells. Conclusions: Our cases illustrate endocrine organ manifestations of this rare disease. The condition may respond to combination chemotherapy.
Sclerotic lipoma: lipomas simulating sclerotic fibroma
Authors: ZELGER B.G. , ZELGER B. , STEINER H. , RÜTTEN A.
Published: November 2003

Aims: We report a series of five unusual subcutaneous lipomas characterized by prominent stromal sclerosis in a storiform arrangement. Methods and results: All lesions occurred in young male adults (23.0  ±  9.5 years), three lesions were on the scalp and two on the hands. Clinically, lipoma, chondroma or some form of cyst were suspected. In contrast to sclerotic fibroma (‘circumscribed storiform collagenoma’) a variable number of adipocytes were interspersed within the fibrosclerotic background. These cells also showed immunoreactivity for S100 protein as well as intracytoplasmic lipid deposits ultrastructurally. None of the lesions recurred within a follow‐up period of between 0.5 and 9 years. Conclusions: Our series suggests that sclerotic lipomas should be classified with fibro‐ and myxolipomas, angiolipomas, spindle cell and pleomorphic lipomas, as an additional characteristic subtype of lipoma. Sclerotic lipomas are often misdiagnosed as a sclerotic fibroma, but are essentially fatty tumours and are, at least in our small series, not associated with Cowden's disease.
Hepatitis C virus liver disease in women infected with contaminated anti‐D immunoglobulin
Authors: SHEEHAN M.M. , DOYLE C.T. , WHELTON M. , KENNY‐WALSH E.
Published: November 2003

Screening for hepatitis C virus (HCV) infection is carried out by detection of antibodies to the virus (enzyme‐linked immunosorbent assay (ELISA) and recombinant immunoblot assay (RIBA)) with confirmation by identification of HCV RNA genome in serum (polymerase chain reaction (PCR)). We describe the histological features on liver biopsy in 88 women with chronic HCV infection (serum positive on ELISA, RIBA and PCR) acquired from virus contaminated anti‐D immunoglobulin. For the majority of these patients the time interval from virus infection to presentation was between 17 and 18 years. We separately assessed necroinflammatory disease activity and architectural features on liver biopsy and applied a scoring system which permitted semi‐quantitative documentation of abnormal features. Only three women showed liver biopsies within normal limits (± focal steatosis). The remaining 85 cases showed a predominantly mild or moderate degree of disease activity with interface hepatitis (56.8% of cases), spotty necrosis, apoptosis and focal inflammation (88.6% of cases) and portal inflammation (90.9% of cases). Confluent necrosis was an uncommon finding (2.3% of cases).

Assessment of architectural features showed normal appearance in 35.2% of biopsies. The predominant architectural abnormality noted was portal tract fibrosis. Ten per cent of cases, however, showed significant fibrous band and/or nodule formation.
Poorly differentiated myoepithelial cell rich carcinoma of the breast
Authors: DAMIANI S. , RICCIONI L. , PASQUINELLI G. , EUSEBI V.
Published: November 2003

Three unusual cases of invasive breast carcinoma are reported, each comprising a dual malignant cellular proliferation consisting of ‘ordinary’ epithelial cells as well as myoepithelial cells haphazardly intermingled. The cases displayed features which did not match any of the four main types of invasive malignant breast tumours containing myoepithelial elements, i.e. adenomyoepithelioma, adenoid cystic carcinoma, pure myoepithelioma and low‐grade adenosquamous carcinoma. The designation of ‘poorly differentiated myoepithelial cell rich carcinoma’ (PDMC) is proposed for these tumours.
Expression of p53, mdm2, p21/waf1 and bcl‐2 proteins in thymomas
Authors: STEFANAKI K. , RONTOGIANNI D. , KOUVIDOU C.H. , BOLIOTI S. , DELIDES G. , PANTELIDAKI A. , SOTSIOU F. , KANAVAROS P.
Published: November 2003

We have investigated the immunohistochemical expression of p53, mdm2, p21/waf1 and bcl‐2 proteins in 31 thymic epithelial tumours comprising five medullary thymomas (MDT), four mixed thymomas (MT), 12 cortical thymomas (CT), eight predominately cortical thymomas (PCT) and two well‐differentiated thymic carcinomas (WDTC). We have found p53, mdm2, p21 and bcl‐2 protein expression in 25/31, 8/31, 5/31 and 10/31 thymic epithelial tumours, respectively. Coexpression of p53 and mdm2 proteins was found in eight cases (three CT, four PCT and one WDTC). Five of them were also p21 positive and three p21 negative. Discordant p53+/mdm2−/p21− protein expression was found in 19 cases (three MDT, three MT, nine CT, three PCT and one WDTC). Mdm2 and p21 proteins were not expressed in the absence of p53 protein. Coexpression of bcl‐2 and p53 proteins was found in seven cases (three MDT, three MT and one WDTC). Eighteen cases were p53+/bcl‐2− (10 CT, seven PCT and one WDTC) and three cases (two MDT and one MT) were bcl‐2+/p53−. Our findings indicate that in thymomas, p53 expression is more frequently associated with cortical histotypes while bcl‐2 expression is strongly associated with medullary and mixed histotypes. In addition, there is an inverse correlation between p53 and bcl‐2 protein expression in thymomas. Coexpression of p53/mdm2/p21 proteins may reflect thymomas with wild‐type (wt), p53 gene since mdm2 and p21 proteins are inducible by wt p53 gene. However, in view of previous findings that p53 mutation is an early event in thymomas, the possibility of p53 gene mutation with p53‐independent mdm2 and p21 expression should be considered in these cases. Discordant p53+/mdm2−/p21− protein expression may represent thymomas with p53 gene mutations unable to activate expression of mdm2 and p21 proteins.
The endometrial deficient secretory phase
Authors: THORNBURGH I. , ANDERSON M.C.
Published: November 2003

The deficient secretory phase is a functional abnormality of the endometrium that has hitherto been poorly recognized. Endometrial curettings from 34 cases were examined in detail by histology. We also performed morphometric analyses of epithelial cell nuclei and assessed the oestrogen and progesterone receptor status of these cases compared with controls. Clinicopathological correlations were examined. The cases showing the deficient secretory phase were characterized histologically by the presence of elongated, hyperchromatic glandular cell nuclei, diminished or no secretory activity in the second half of the menstrual cycle and poorly developed stroma. Morphometry confirmed that the nuclei were different in shape from those seen at any time during the normal menstrual cycle and from basal endometrium. Nuclear expression of oestrogen and progesterone receptors in formalin‐fixed and paraffin‐embedded sections was reduced. It is apparent from this study that the endometrial appearances described represent a definable condition that may be linked to menstrual irregularities and, in some circumstances, infertility; it should be more widely recognized.
Myositis ossificans‐like lesion of nerve
Authors: WASMAN J.K. , WILLIS J. , MAKLEY J. , ABDUL‐KARIM F.W.
Published: November 2003

An unusual ossifying lesion arising within the median nerve of a young adult male is presented. While ectopic ossification has been described in a large number of sites, intraneural ossification is an uncommon event and usually occurs in association with a pre‐existing intraneural neoplasm. This case, in contrast, bears striking histopathological resemblance to myositis ossificans with prominent zonation. We believe that this may represent the first example of a benign myositis ossificans‐like lesion of nerve.
Mitogenic stimulation of T cells reveals differing contributions for B7‐1 (CD80) and B7‐2 (CD86) costimulation
Authors: PERRIN P. J. , DAVIS T. A. , SMOOT D. S. , ABE R. , JUNE C. H. , LEE K. P.
Published: November 2003

The requirement of accessory cells for concanavalin A (Con A) activation of T cells suggests delivery of a separate costimulatory signal. However, the costimulatory pathways involved have not been identified. These studies assess the role of CD28–B7‐mediated costimulation during T‐cell activation by Con A. The B7‐1/B7‐2 binding protein CTLA4‐Ig inhibited the proliferative response of primary lymph node cells to either Con A or soluble anti‐CD3 mAb. This suppression was dose dependent and could be reversed by CD28 cross‐linking. CTLA4‐Ig also completely suppressed induction of interleukin‐2 (IL‐2) mRNA by Con A. CTLA4‐Ig‐mediated suppression was not due to blockade of the Con A ‘receptor(s)’ or of the primary activation signal (as measured by the intracellular calcium response). Although both B7‐1 and B7‐2 were up‐regulated following Con A activation, each played a different role in proliferation and cytokine production. Individually, anti‐B7‐2 Fab partially inhibited the Con A response whereas anti‐B7‐1 Fab had no effect. However, the combination of anti‐B7‐1 and anti‐B7‐2 Fab completely suppressed proliferation and IL‐2 production. Therefore, while a part of the Con A response requires B7‐2, the remainder of the response can utilize either B7‐1 or B7‐2. Together, these results demonstrate that Con A activation of T cells requires the delivery of a separate costimulatory signal that is mediated almost entirely by the B7 receptors.
Thrombin stimulates production of interleukin‐8 in human umbilical vein endothelial cells
Authors: UENO A. , MURAKAMI K. , YAMANOUCHI K. , WATANABE M. , KONDO T.
Published: November 2003

Interleukin‐8 (IL‐8) is regarded as an important mediator of inflammation because of its potent and specific chemotactic activity on neutrophils. In the present investigation, human umbilical vein endothelial cells (HUVEC) stimulated with thrombin were found to produce IL‐8, in a dose‐ and time‐dependent manner. After stimulation with 10 U/ml thrombin for 24 hr, the level of IL‐8 in the conditioned medium was 14 ng/ml, or enough to elicit PMN chemotaxis in vitro. Northern blot analysis revealed that thrombin as well as IL‐1β elevates the level of IL‐8 mRNA preceding the formation of IL‐8 protein. A synthetic peptide SFLLRN [human thrombin receptor‐activating peptide (TRAP)] was found to mimic the action of thrombin. Preincubation with anti‐thrombin compounds such as hirudin and antithrombin‐III–heparin almost completely suppressed the action of thrombin without affecting the actions of other stimuli including IL‐1β, phorbol 12‐myristate 13‐acetate (PMA) and TRAP. Diisopropylfluorophosphate‐treated thrombin did not stimulate IL‐8 production. Calphostin‐C, a protein kinase C (PKC) inhibitor, attenuated the production of IL‐8 by thrombin, TRAP and PMA, but left the action of IL‐1β unchanged. These results strongly suggest that catalytic activation of thrombin receptor by thrombin results in PKC‐dependent IL‐8 production accompanied by an increase in IL‐8 mRNA level.
Production of granulocyte–macrophage colony‐stimulating factor by T cells is regulated by B7 and IL‐1β
Authors: KRUGER M. , VAN GOOL S. , PENG X. H. , COOREVITS L. , CASTEELS‐VAN DAELE M. , CEUPPENS J. L.
Published: November 2003

Granulocyte–macrophage colony‐stimulating factor (GM‐CSF) has proliferation‐ and differentiation‐inducing effects on immature myeloid cells in the bone marrow, and it can modulate the function of several types of mature myeloid cells. We have stimulated purified human T cells with immobilized anti‐CD3 or mitogenic anti‐CD2 (a combination of monoclonal antibodies 9‐1 and 9.6) which could induce GM‐CSF production. The cytokines interleukin‐1β (IL‐1β) and IL‐2 strongly enhanced GM‐CSF production, while IL‐4, IL‐6, GM‐CSF, interferon‐γ (IFN‐γ) and tumour necrosis factor (TNF) had no effect. Activation of protein kinase C by phorbol myristate acetate or triggering of CD28 on T cells by monoclonal antibody 9.3 provided accessory signals for enhanced GM‐CSF production in activated T cells. Most important, the addition of mouse cells transfected with human B7‐1 (CD80), a natural ligand for CD28, provided a potent accessory signal for GM‐CSF production by activated T cells, which could not be blocked by cyclosporin A. The effect of IL‐1β was in fact indirect, and resulted from enhanced IL‐2 production, while the effect of B7 resulted from both IL‐2‐dependent and IL‐2‐independent pathways. We conclude that antigen‐presenting cells (APC) can up‐regulate GM‐CSF production through IL‐1β and through CD28 triggering by B7 molecules. As GM‐CSF itself up‐regulates B7 expression and IL‐1β production by APC, a bidirectional regulatory feedback pathway between APC and T cells seems to modulate GM‐CSF production.
Chemical carcinogens and antigens induce immune suppression via Langerhans' cell depletion
Authors: WOODS G. M. , QU M. , RAGG S. J. , MULLER H. K.
Published: November 2003

The ability of the chemical carcinogen dimethylbenz(a)anthracene (DMBA) to deplete Langerhans' cells (LC) from murine skin is crucial to the development of antigen‐specific suppression. This depletion is a consequence of the LC recognizing the DMBA as antigenic and migrating to the draining lymph nodes to attempt to elicit T‐cell activation. This depletion also occurred following exposure to high doses of the contact sensitizers 2,4‐dinitrofluorobenzene (DNFB), 2,4,6‐trinitrochlorobenzene (TNCB) and fluorescein isothiocyanate (FITC). However, LC depletion was not significant at lower doses, even though these doses were sufficient to induce strong contact sensitivity responses. Application of the contact sensitizer, DNFB, through skin depleted of LC (by pretreatment with either the carcinogen DMBA or the antigen TNCB) failed to induce contact sensitivity. This immune non‐responsiveness was antigen specific, and could be transferred by spleen cells to naive mice, which were unable to respond to DNFB. Mouse skin treated with doses of TNCB, that did not cause LC depletion but still induced a normal contact hypersensitivity, retained its ability to initiate a normal immune response to DNFB. Together these findings demonstrate that carcinogens share some properties with antigens as they both cause LC depletion and interact with the immune system. Furthermore, it is this LC depletion, rather than carcinogen treatment, that is a critical factor which leaves the skin immunologically compromised and favours the induction of antigen‐specific suppression.
Antigen presentation in the murine oral epithelium
Authors: ERIKSSON K. , AHLFORS E. , GEORGE‐CHANDY A. , KAISERLIAN D. , CZERKINSKY C.
Published: November 2003

We have previously reported that the buccal mucosa can support delayed type hypersensitivity (DTH) reactions to contact sensitizers. In the present study, we show that cells isolated from the buccal epithelium are able to present soluble exogenous antigens to specific T cells. Single cell suspensions obtained by enzymatic dispersion of buccal epithelial sheets could present the native protein antigen hen‐egg lysozyme (HEL) to the I‐Ak‐restricted CD4+ T‐cell hybridoma specific for a.a 46–61 on HEL. T‐cell activation resulted in interleukin‐2 (IL‐2) production which could be inhibited by anti‐major histocompatibility complex (MHC) class‐II antibodies of pertinent specificity. Immunohistochemical staining of whole buccal epithelial sheets revealed that all MHC II positive cells had a dendritic morphology and expressed ATPase activity, indicating that these cells represent a major antigen‐presenting cell (APC) population in this tissue. Furthermore, single cell suspensions isolated from buccal epithelium (BEC) after local in vivo administration of either a native soluble protein, a synthetic dodecapeptide, or a contact sensitizer were able to activate antigen‐specific T cells ex vivo. Kinetic analyses indicated that maximal APC activity in the oral epithelium occurred within 1 hr after local antigen administration, and had essentially vanished after 24 hr. Conversely, APC activity was undetectable in draining cervico‐mandibular lymph node cell suspensions recovered 1 hr after local antigen injection but became manifest after 3–24 hr. These observations suggest that dendritic cells can acquire antigens in the buccal epithelium and migrate to draining lymph nodes where they present processed antigen to MHC class II‐restricted T cells. This APC population may thus be a critical element in the initiation of Th1‐driven DTH responses in the oral mucosa.
HIV‐1 envelope glycoprotein gp120 down‐regulates CD4 expression in primary human macrophages through induction of endogenous tumour necrosis factor‐α
Authors: KARSTEN V. , GORDON S. , KIRN A. , HERBEIN G.
Published: November 2003

Among immunological abnormalities present in human immunodeficiency virus type 1 (HIV‐1)‐infected individuals are dysregulation of cytokine production and CD4 down‐regulation in both T‐helper cells and monocytes/macrophages. The HIV‐1 envelope glycoprotein 120 (gp120) has the ability to induce different cytokines in peripheral blood mononuclear cells and in monocytes/macrophages in vitro which in some instances have been reported to down‐regulate macrophage CD4 expression. This study provides evidence that HIV‐1 recombinant gp120 (rgp120) down‐regulates both surface and total CD4 expression in primary tissue culture‐differentiated macrophages (TCDM) at the level of transcription. The CD4 down‐regulation observed in TCDM occurred between 6 and 12 hr after rgp120 treatment preceded by a peak of endogenous tumour necrosis factor‐α (TNF‐α) observed at 3–6 hr post‐treatment. We demonstrate that the TCDM CD4 down‐regulation observed after rgp120 treatment was inhibited by the use of an anti‐huTNF‐α monoclonal antibody (mAb), but not by mAb directed against other cytokines induced by rgp120, such as interleukin‐1β (IL‐1β) and interferon‐α (IFN‐α). The present findings roughly parallel those observed both in the sera of patients and in the monocytes/macrophages isolated from HIV‐positive individuals, suggesting that gp120 by stimulating endogenous TNF‐α production could be a good candidate for the CD4 down‐regulation observed in the monocytes/macrophages of HIV‐1‐infected individuals. In contrast to CD4 down‐regulation in HIV‐infected lymphocytes, which results from a direct effect of viral genes on CD4 expression, soluble factors such as cytokines induced during HIV infection might explain the monocyte/macrophage CD4 dysregulation observed in acquired immune deficiency syndrome.
Catecholamines are synthesized by mouse lymphocytes and regulate function of these cells by induction of apoptosis
Authors: JOSEFSSON E. , BERGQUIST J. , EKMAN R. , TARKOWSKI A.
Published: November 2003

The immune and the nervous systems are anatomically closely related and interact with each other by molecules common to both systems, such as cytokines and neurotransmitters. The purpose of this study was to investigate the participation of catecholamines in the neuroimmunological network. The ability of immune cells to produce catecholamines was examined by a highly sensitive capillary electrophoresis assay, which permits detection of easily oxidized catecholamines in the zeptomole (10−21) range. In addition, the effects of catecholamines on in vitro proliferation, differentiation and apoptosis of lymphocytes were assessed. Mouse spleen cells and macrophages contained on average 7 × 10−17 and 2 × 10−17 mole dopamine per cell, respectively. In the former cell population also norepinephrine was found. Several mouse B‐ and T‐cell hybridomas were also shown to contain endogenously produced dopamine in levels ranging from 7 × 10−20 to 2 × 10−18 mole dopamine per cell. In addition, one of the T‐cell hybridomas proved to synthesize norepinephrine. The dopamine production of lymphocytes was blocked by the tyrosine hydroxylase inhibitor α‐methyl‐p‐tyrosine, whereas incubation with the precursor L‐DOPA increased the dopamine content. Incubation with L‐DOPA, dopamine and norepinephrine dose‐dependently suppressed mitogen induced proliferation and differentiation of mouse lymphocytes. Even short‐time pretreatment of lymphocytes with L‐DOPA and dopamine strongly suppressed lymphocyte proliferation and cytokine production. Incubation of lymphoid cells with L‐DOPA, dopamine and norepinephrine dose‐dependently induced apoptosis which, at least partly, explains the suppressive effects of catecholamines on lymphocyte function. Our results demonstrate that catecholamines: (i) are actively produced by lymphocytes and (ii) have the capacity to act as auto‐ and/or paracrine regulators of lymphocyte activity through induction of apoptosis.
Heat‐shock protein expression on the membrane of T cells undergoing apoptosis
Authors: POCCIA F. , PISELLI P. , VENDETTI S. , BACH S. , AMENDOLA A. , PLACIDO R. , COLIZZI V.
Published: November 2003

Heat‐shock proteins (hsp) represent a highly conserved family of proteins, normally localized in the cytoplasm and nucleus, whose expression is induced in situations involving cell stress. This paper reports the unusual translocation of hsp to the cell membrane of T cells undergoing apoptosis. We observed that glucocorticosteroid‐induced thymocyte death is associated to the surface expression of hsp  60 and hsp  70 in a discrete fraction of apoptotic cells. hsp surface expression is closely related to a thymic subset of immature CD3low/− T cells. The expression of surface hsp  60 appears early after treatment with dexamethasone (3 hr) whereas the membrane expression of hsp  70 follows different kinetics and peaks later. Morphological analysis of the hsp+ apoptotic cells suggest that this subset represents late‐stage apoptotic cells at their minimal volume before fragmentation into apoptotic bodies. Membrane expression of hsp is also associated with apoptosis in peripheral blood mononuclear cells from AIDS patients cultured in vitro. Altogether, we show that a discrete fraction of cells undergoing apoptosis expresses membrane hsp  60 and hsp  70, supporting the hypothesis that apoptosis causes a radical alteration in the expression of cell surface molecules. Surface hsp expressed during apoptosis may constitute a novel immune‐context able to generate packages of self‐ and exogenous antigens, originating from degradation of altered cells.
Analysis of the local kinetics and localization of interleukin‐1α, tumour necrosis‘qc factor‐α and transforming growth factor‐β, during the course of experimental pulmonary tuberculosis
Authors: HERNANDEZ‐PANDO R. , OROZCO H. , ARRIAGA E.K. , SAMPIERI A. , LARRIVA‐SAHD J. , MADRID‐MARINA V.
Published: November 2003

A mouse model of pulmonary tuberculosis induced by the intratracheal instillation of live and virulent mycobacteria strain H37‐Rv was used to examine the relationship of the histopathological findings with the local kinetics production and cellular distribution of tumour necrosis factor‐α (TNF‐α), interleukin‐1α (IL‐1α) and transforming growth factor‐β (TGF‐β). The histopathological and immunological studies showed two phases of the disease: acute or early and chronic or advanced. The acute phase was characterized by inflammatory infiltrate in the alveolar–capillary interstitium, blood vessels and bronchial wall with formation of granulomas. During this acute phase, which lasted from 1 to 28 days, high percentages of TNF‐α and IL‐1α immunostained activated macrophages were observed principally in the interstium‐intralveolar inflammatory infiltrate and in granulomas. Electron microscopy studies of these cells, showed extensive rough endoplasmic reticulum, numerous lysosomes and occasional mycobacteria. Double labelling with colloid gold showed that TNF‐α and IL‐1α were present in the same cells, but were confined to separate vacuoles near the Golgi area, and mixed in larger vacuoles near to cell membrane. The concentration of TNF‐α and IL‐1α as well as their respective mRNAs were elevated in the early phase, particularly at day 3 when the bacillary count decreased. A second peak was seen at days 14 and 21–28 when granulomas appeared and evolved to full maturation. In contrast, TGF‐β production and numbers of immunoreactive cells were low in comparison with the advanced phase of the disease. The chronic phase was characterized by histopathological changes indicative of more severity (i.e. pneumonia, focal necrosis and extensive interstitial fibrosis) with a decrease in the TNF‐α and IL‐1α production that coincided with the highest level of TGF‐β. The bacillary counts were highest as the macrophages became large, vacuolated foamy cells, and containing numerous bacilli with immunoreactivity to mycobacterial lipids and lipoarabinomannan (LAM). These macrophages displayed poor and scarce TNF‐α and IL‐1α immunostaining but still strong immunoreactivity to TGF‐β. These cytokine production kinetics and the spatial relationship between immunostained cells and lung lesions corroborate the important role of TNF‐α and IL‐1α in the constitution of granulomas and immune protection during the early phase of the infection, and also suggest an important if not primary role for TGF‐β in the immunopathogenesis of the advanced forms of pulmonary tuberculosis.
Mechanisms of complement resistance induced by non‐lethal complement attack and by growth arrest
Authors: MARCHBANK K. J. , VAN DEN BERG C. W. , MORGAN B. P.
Published: November 2003

Non‐lethal complement (C) attack on K562 cells has been shown to induce a transient resistance to lethal amounts of C. We have previously shown that incubation of K562 with phorbol 12‐myristate 13‐acetate (PMA) caused an increase in both CD59 expression and resistance to C killing and we were interested to examine whether non‐lethal C attack caused a similar effect. We here demonstrate that expression of the C inhibitors decay‐accelerating factor (DAF), membrane cofactor protein (MCP) and CD59 was unaltered on K562 after non‐lethal C attack and that neutralization of these inhibitors with specific blocking antibodies did not reverse the induced resistance. In an effort to understand the mechanisms of resistance we searched for other conditions that might induce C resistance in K562 cells. Growth‐arrested cells showed a similar degree of resistance to C killing. The levels of DAF and MCP on these cells were unaltered whereas expression of CD59 was markedly reduced. Non‐lethal C attack on these growth‐arrested cells induced a further increase in resistance to C killing, suggesting that the mechanisms of resistance were not identical. Indeed, resistance of non‐lethally attacked cells was completely lost within 8 hr of attack whereas resistance of growth‐arrested cells was detectable for up to 48 hr after returning to cell cycle. These data demonstrate that C resistance induced by two distinct strategies is not mediated by the known membrane C inhibitors. Resistance may be a result of the expression of a novel inhibitor or due to metabolic depletion, a likely common consequence of non‐lethal C attack and induction of growth arrest, implying that cells take an active role in C‐mediated killing.
Eosinophilia, parasite burden and lung damage in Toxocara canis infection in C57Bl/6 mice genetically deficient in IL‐5
Authors: TAKAMOTO M. , OVINGTON K. S. , BEHM C. A. , SUGANE K. , YOUNG I. G. , MATTHAEI K. I.
Published: November 2003

C57Bl/6 mice genetically deficient in interleukin (IL)‐5 (IL‐5−/−) and mice with the normal IL‐5 gene (IL‐5+/+) were infected with embryonated eggs of Toxocara canis. IL‐5+/+ mice developed a marked eosinophilia in their peripheral bloods and bone marrows after infection. In contrast, the number of eosinophils at these sites actually decreased during the acute phase of infection in IL‐5−/− mice. A smaller number of eosinophils infiltrated the lung, liver, heart and skeletal muscle of infected IL‐5−/− mice than those of infected IL‐5+/+ mice. Eosinophils were not produced incultures of bone marrow cells from either IL‐5+/+ or IL‐5−/− mice which were stimulated with excretory–secretory antigen of T. canis larvae. The capacity of cells from the bone marrow to differentiate into eosinophils when stimulated in vitro with recombinant murine IL‐5 was the same whether the cells were from IL‐5+/+ or IL‐5−/− mice. Taken together, these results show that an IL‐5‐like molecule is not produced by the T. canis larvae and that IL‐5 produced by host cells is solely responsible for the eosinophilia in mice infected with this nematode. The number and location of T. canis larvae were not altered in the absence of IL‐5. In contrast, lung damage in infected IL‐5−/− mice was less extensive than that in infected IL‐ 5+/+ mice, although structures resembling Charcot–Leyden crystals were seen in the lungs of both IL‐5+/+ and IL‐5−/− mice. These results suggest that eosinophils play a role in the pathology in mice infected with T. canis.
Calcium pyrophosphate dihydrate crystals activate MAP kinase in human neutrophils: inhibition of MAP kinase, oxidase activation and degranulation responses of neutrophils by taxol
Authors: JACKSON J. K. , TUDAN C. , SAHL B. , PELECH S. L. , BURT H. M.
Published: November 2003

The activation of MAP kinase in human neutrophils stimulated by both uncoated and plasma‐opsonized crystals of triclinic calcium pyrophosphate dihydrate (CPPD) was investigated. The effect of taxol on MAP kinase activation and on the responses of neutrophils stimulated by plasma‐opsonized crystals was determined. MAP kinase activation was identified and quantified in Mono Q chromatography separated fractions of neutrophils that had been incubated with CPPD crystals by measuring []> γ‐32P]adenosine triphosphate (ATP) phosphorylation of myelin basic protein and using immunoblotting techniques. Human neutrophils were incubated with taxol (0–50 μm), added to plasma‐opsonized CPPD (50 mg/ml) and MAP kinase activation, chemiluminescence, superoxide anion generation, lysozyme and myeloperoxidase release were monitored. Both uncoated and plasma coated CPPD crystals induced a large increase in MAP kinase activity in neutrophils over control levels within 1 min of incubation. Pretreament of neutrophils with taxol was able to suppress this activation of MAP kinase. Taxol produced a concentration‐dependent inhibition of opsonized CPPD‐induced neutrophil chemiluminescence, superoxide anion production and myeloperoxide release. Taxol at 28 μm also significantly inhibited chemiluminescence, superoxide anion production and myeloperoxidase release from neutrophils stimulated by opsonized zymosan. This is the first report of crystal‐induced activation of MAP kinase in neutrophils. Microtubule‐associated processes, such as signal transduction, secretion and phagocytosis are involved in particulate‐induced neutrophil responses. We have suggested that the inhibitory effect of taxol observed in this work is due to its stabilizing effect on microtubules and disruption of MAP kinase activation associated with microtubules.
Preliminary results and analysis of a new set of sonic instruments for root‐end cavity preparation
Authors: von Arx T. , Kurt B. , Ilgenstein B. , Hardt N.
Published: November 2003

The introduction of microinstruments for root‐end cavity preparation has clearly improved the surgical technique in periradicular surgery with apicectomy. The new generation of sonic and ultrasonic root‐end preparation devices, usually termed ‘retrotips’, has simplified the preparation of a sufficiently deep cavity that follows the original path of the root canal. The present paper describes the use of a new set of diamond‐surfaced retrotips driven by a sonic handpiece. For clinical evaluation a prospective study was carried out, with assessment of pre‐, intra‐ and postoperative data in 50 teeth (43 patients). This paper analyses: (i) the applicability of the new retrotips; (ii) the quality of the root‐end filling assessed radiographically; (iii) the immediate postoperative course. Access with the retrotips to the resected root‐end was found to be excellent in 80% of the treated teeth. The postoperative radiographs showed a root‐end filling of at least 3 mm in 70% of the resected teeth. Healing was uneventful in all cases and most patients presented without any symptoms after 5 days and 10 days. The long‐term treatment outcome will be reported in a future paper.
A methacrylate‐based cement used as a root canal sealer
Authors: Ahlberg K. M. F. , Tay W.‐M.
Published: November 2003

In this study, a methacrylate‐based orthopaedic bone cement was modified for use as a root canal sealer by changing the monomer from n‐butyl methacrylate to tetrahydrofurfuryl methacrylate. The sealing and bonding capabilities, solubility and handling properties of the resin were evaluated. Seventy‐three extracted mature human teeth with single canals were used. The teeth were randomly divided into two experimental groups each of 30 teeth; three control teeth were set aside for the leakage tests. Ten teeth were reserved for the scanning electron microscope studies. All the experimental teeth were root filled with gutta‐percha and the resin sealer. The particle size of the polymer was limited in Group 1 to 75 μm or less and in Group 2 to 45 μm or less. The controls were filled with gutta‐percha only. Silver nitrate solution was used as the leakage indicator. In Group 1 the mean linear penetration of the indicator was 1.10 mm ± 1.04; four roots showed no leakage. In Group 2 the mean linear penetration of the indicator was 0.57 mm ± 0.65; 12 roots showed no leakage. The test bodies showed no weight change after 1 and 3 months' storage in saline at 37°C. The scanning electron microscope examination revealed a bond both between the resin and the dentine and between the resin and the gutta‐percha. The working time of the new resin composition was normally about 50 min, and handling was found to be easy.
Clinical study of refractory apical periodontitis treated by apicectomy Part 1. Root canal morphology of resected apex
Authors: Wada M. , Takase T. , Nakanuma K. , Arisue K. , Nagahama F. , Yamazaki M.
Published: November 2003

The morphology of the root apex was analysed by observation of the anatomy of specimens obtained by apicectomy in cases of refractory apical periodontitis that did not respond to nonsurgical root canal treatment. Apical ramifications were present in 19 (70%) of the roots, while one were found in the remaining eight (3%) roots. This frequency is far higher than that reported by other investigators, suggesting that there is a close relationship between the anatomical complexity of the root canal and the occurrence of refractory apical periodontitis.
Three‐dimensional reconstruction of Widmanstätten plates in Fe–12.3Mn–0.8C
Authors: MANGAN M. A. , LAUREN P. D. , SHIFLET G. J.
Published: November 2003

The three‐dimensional reconstruction of serial sections is applied to examine Widmanstätten plates in a high‐manganese steel. Serial sections were obtained through repeated metallographic polishing and imaging and interpolation was done using the ‘watershed’ algorithm. Electron backscatter diffraction was used to determine grain misorientations and cementite:austenite orientation relationships. This technique yields a unique combination of information concerning crystallography and the three‐dimensional development of a solid body.
In vitro suppression of fungal root pathogens of cereals by Brassica tissues
Authors: KIRKEGAARD J. A. , WONG P. T. W. , DESMARCHELIER J. M.
Published: November 2003

The superior growth of wheat following Brassica crops compared to that following non‐Brassica crops may be due to the suppression of soilborne fungal pathogens by volatile isothiocyanates (ITCs) released in the soil during hydrolysis of glucosinolates contained in Brassica tissues. We investigated the effects of volatile compounds released from the root, shoot and seed meal tissues of canola (Brassica napus) and Indian mustard (Brassica juncea) on the mycelial growth of five soilborne pathogens of cereals—Gaeumannomyces graminis var. tritici, Rhizoctonia solani, Fusarium graminearum, Pythium irregulare and Bipolaris sorokiniana. Three isolates of each species, originally collected from the roots of wheat (Triticum aestivum) and barley grass (Hordeum leporinum ) in southern Australia, were exposed to volatiles released in vitro when sterile water was added to freeze‐dried Brassica tissues. The root and shoot tissues of both Brassica species were more suppressive at flowering than maturity and mustard tissues were generally more suppressive than canola. The degree of fungal suppression by the various Brassica tissues was related to the concentration and type of isothiocyanates released, which varied with Brassica species, tissue age and tissue type. There were significant differences in the sensitivity of the fungal species and among isolates of each species. Gaeumannomyces and Rhizoctonia were generally the most sensitive to the volatiles released, Pythium and Bipolaris the least. The results indicate that the effectiveness of fungal suppression by Brassica crops will depend upon the species, age and type of Brassica tissue, which influence the type and concentration of isothiocyanates evolved, and the sensitivity of the pathogen.
Biocontrol of sclerotinia stem rot (Sclerotinia minor) in sunflower by seed treatment with Gliocladium virens
Authors: BURGESS D. R. , HEPWORTH G.
Published: November 2003

An isolate of Gliocladium virens from disease‐affected soil in a commercial sunflower field proved highly antagonistic to Sclerotinia minor in culture. Significant disease control was obtained when a conidial suspension was applied over a limited area of potting medium and lateral roots were inoculated outside the treated area at the edge of the pot. Most promising, in terms of minimizing the amount of G. virens required for disease control, was the significant bioprotection obtained by seed treatment. In glasshouse experiments following inoculation of lateral roots with S. minor at 6–7 weeks after sowing, disease incidence was reduced from 60.0% to 15.8% in potting medium and from 66.7% to 12.5% in field soil. Seed treatment was also effective in field microplots, where disease incidence was reduced from 37.0% to 22.2%. G. virens applied on seed did not affect the dry weight of the lateral root system.
Seasonal changes in pathotype complexity in French populations of barley powdery mildew
Authors: CAFFIER V. , HOFFSTADT T. , LECONTE M. , DE VALLAVIEILLE‐POPE C.
Published: November 2003

Airborne conidia of Erysiphe graminis f.sp. hordei were sampled in three regions and a single locality in the northern part of France for 2 years. Sampling was carried out in early spring, in late spring and in autumn, in order to separate the effects of winter barley cultivars, carrying few specific resistance alleles, and of spring barley cultivars, carrying diverse resistance alleles, on the structure of the pathogen population. Although complex pathotypes with three to 10 virulences were selected by spring cultivars, simple pathotypes, including a pathotype with the single unnecessary virulence allele Va22, which formed a clear majority of the samples, remained dominant in early spring, when winter but not spring cultivars were growing. In early spring, simple pathotypes were more prevalent in the north, where the winter cultivars represented 90% of the barley acreage, than in the east, where winter cultivars represented 65%. In the west, the frequency of simple pathotypes was limited compared to the north, possibly because of the resistance allele Mlg in winter cultivars. The high frequency of simple pathotypes in early spring could be explained by a differential adaptation between simple and complex pathotypes or by delayed epidemics on spring cultivars compared to winter cultivars.
Isoenzyme diversity in Xanthomonas campestris pv. mangiferaeindicae
Authors: SOMÉ A. , SAMSON R.
Published: November 2003

A collection of 26 strains of Xanthomonas campestris pv. mangiferaeindicae isolated from three different host species in eight countries was investigated for variation in isozyme patterns. Three enzyme systems were analysed: esterase (EST), phosphoglucomutase (PGM) and superoxide dismutase (SOD). Four groups of strains were identified: nonpigmented strains isolated from mango and pepper‐tree in Australia, Comores, India, Reunion Island, South Africa, and Taiwan; nonpigmented Brazilian strains from mango; nonpigmented strains from ambarella isolated in the French West Indies; heterogeneous yellow pigmented strains from mango (Brazil and Reunion Island). The value of isozyme profiling as markers of the pathogenicity groups in X. c. pv. mangiferaeindicae is discussed.
PCR amplification of the fas‐1 gene for the detection of virulent strains of Rhodococcus fascians
Authors: STANGE R. R. , JEFFARES D. , YOUNG C. , SCOTT D. B. , EASON J. R. , JAMESON P. E.
Published: November 2003

Thirty‐six isolates of the fasciation‐inducing bacterium Rhodococcus fascians were evaluated for the presence and location of the fas‐1 gene, which codes for an isopentenyl transferase, the committed step in cytokinin biosynthesis. The presence of fas‐1 was determined by PCR using a set of primers to the most conserved regions of the gene and by Southern hybridization to genomic digests using the PCR fragment as a probe. Both methods revealed the presence of the gene in 18 virulent isolates and the absence of the gene in 18 avirulent isolates. Thus, there is a strong relationship between the presence of the gene and virulence of the organism. The location of fas‐1 was determined by probing blots of linear and circular DNA. For most of the virulent isolates, the gene was localized to a 200 ± 10 kb linear plasmid. Three virulent isolates lacked a plasmid of this size, but contained fas‐1 either on a linear plasmid of 130 kb or on a large circular plasmid.
Germination of Alternaria linicola conidia on linseed: effects of temperature, incubation time, leaf wetness and light regime
Authors: VLOUTOGLOU I. , FITT B. D. L. , LUCAS J. A.
Published: November 2003

Conidia of Alternaria linicola germinated on both water agar and linseed leaves (detached or attached) over a wide range of temperatures (5–25°C) by producing one to several germ tubes. At temperatures between 10°C and 25°C and under continuous wetness in darkness, germination started within 2 h after inoculation and reached a maximum (100%) by 8 to 24 h, depending on temperature. At 5°C, the onset of germination was later and the rate of germ tube elongation was slower than that at 10–25°C. During germination, conidia of A. linicola were sensitive to dry interruptions of wet periods and to light. Short (2 h) or long (12 h) dry interruptions occurring at any time between 2 and 6 h after inoculation stopped conidial germination and germ tube elongation. With continuous wetness, light periods 2 to 12 h long immediately after inoculation inhibited conidial germination, which was resumed only when a dark period followed subsequently. However, germination and germ tube elongation of A. linicola conidia stopped and the viability of the conidia was lost during exposure to dry light periods immediately after inoculation with spore suspensions. Penetration of leaves by A. linicola was evident after 12 h and occurred mainly through epidermal cells (direct) with or without the formation of appressoria.
Genetic comparison of the peach yellow leaf roll agent with European fruit tree phytoplasmas of the apple proliferation group
Authors: KISON H. , KIRKPATRICK B. C. , SEEMÜLLER E.
Published: November 2003

Restriction fragment length polymorphism (RFLP) analysis of PCR‐amplified ribosomal DNA and Southern blot hybridization using cloned chromosomal DNA fragments from the apple proliferation (AP) phytoplasma as probes were used to investigate the genetic relationship of the California peach yellow leaf roll (PYLR) agent with phytoplasmas causing fruit tree diseases in Europe. This comparison showed that the California PYLR phytoplasma is closely related to apple proliferation (AP), pear decline, and European stone fruit yellows phytoplasmas and that it is a member of the phylogenetic AP group. The PYLR agent could clearly be distinguished from the AP and European stone fruit yellows phytoplasmas by Southern blot hybridization with DNA fragments from the AP phytoplasma and by RFLP analysis of ribosomal DNA employing SspI, BsaAI, and RsaI restriction endonucleases. However, the PYLR phytoplasma was indistinguishable from the pear decline agent by RFLP analysis of PCR‐amplified ribosomal DNA.