bluuebunny/crossref_metadata_2025 huggingface dataset:
Daycare diagnostic laparoscopy: experience with the first 1000 patients in a tropical gynaecological service
Authors: Babarinsa I.A. , Ajayi A.B. , Adewole I.F. , Adeleye J.A.
Published: November 2003

Objective

To review outpatient diagnostic laparoscopy in a tropical gynaecological service.

Design

A retrospective, descriptive study.

Setting

A teaching hospital in Nigeria, West Africa.

Subjects

Patients in the reproductive age group, predominantly with symptoms of infertility and chronic pelvic pain.

Main outcome measures

The safety and acceptability of outpatient diagnostic laparoscopy, especially as related to the most common complications, the status of the laparoscopist and instances where the procedure initially could not be performed satisfactorily.

Results

Over a period of 10 years, 1033 patients underwent laparoscopy. Most procedures were performed by senior registrars (46.4%). The most common associated problem (22.7%) was allergic cutaneous reaction to pethidine. Initially, laparoscopy could not be performed in 4% of patients, presumably due to inadequate analgesia. Failure to achieve a pneumoperitoneum was, however, the main technical problem associated with a primary patient drop‐out rate of 2.32%.

Conclusion

Daycare diagnostic laparoscopy appears to be acceptable in tropical gynaecological practice. It is recommended for more widespread use with a view to saving the patient's time, reducing costs and minimizing anaesthetic risks.
Laparoscopic Burch colposuspension for urinary stress incontinence: results from 109 consecutive cases
Authors: Lam A. M. , Rosen D. M. B.
Published: November 2003

Objectives To assess the technical feasibility, safety, potential benefits and outcome of laparoscopic Burch colposuspension. Methods Between 1 June 1993 and 1 June 1996 109 women who presented with urodynamically confirmed urinary stress incontinence were prospectively assessed and underwent laparoscopic Burch colposuspension. Results The operation was successfully completed laparoscopically in 107 women. One case was converted to laparotomy due to injury to the inferior epigastric artery and another was precluded from the study due to unexpected intra‐abdominal metastatic disease found at laparoscopy. A total of 19 patients had undergone previous unsuccessful anti‐incontinence surgery. The overall success rate was 98.2% with follow‐up duration between 6 weeks and 36 months (mean 16 months). The overall incidence of perioperative complications was 11.1%. The average time required to perform colposuspension alone was 40 min. The operation was associated with minimal blood loss and postoperative analgesic requirements. The average length of hospital stay was 2.3 days, and the average postoperative bladder catheterization was 1.5 days. Most patients were able to resume normal lifestyle within 1–2 weeks. Conclusion These results confirm the findings of an earlier study that laparoscopic Burch colposuspension is a safe and technically feasible alternative to the open technique. The laparoscopic approach offers the surgeon excellent view and access to the operating field, and the patients the benefits of shortened postoperative catherization time, hospital stay and early return to normal activities. Follow‐up to date indicates a success rate comparable to the traditional open technique.
Laparoscopic extraction of a calcified autoamputated ovary
Authors: Abd‐el‐Maeboud K. H. I. , Salama A.
Published: November 2003

A case of ovarian autoamputation, as a result of torsion of the adnexa, in a 23‐year‐old infertile patient is presented. To our knowledge, this is the first case with this rare diagnosis to be managed via laparoscopy.
The repair of intrinsic sphincter dysfunction with hypermobile urethra by the laparoscopic Burch–Ball procedure
Authors: Ross J. W.
Published: November 2003

Treatment of intrinsic sphincter dysfunction in association with genuine stress incontinence has a high surgical failure rate. The Ball modification of the laparotomy Burch colposuspension has been used with a high degree of success by some authors. In the present study, 11 patients were shown to have genuine stress incontinence by transperineal ultrasound, cough stress test, and multichannel urodynamics. Each subject had a hypermobile urethra, an abnormal valsalva leak‐point pressure, and a low urethral closure pressure. A laparoscopic Burch–Ball colposuspension was performed and patients were then re‐evaluated at 6 weeks and 1 year. There was a 90% objective cure rate at 1 year and the hospital stay and the postoperative recovery time was short. This preliminary study suggests that the laparoscopic procedure is as effective as laparotomy.
Immunohistochemical phenotype of malignant mesothelioma: predictive value of CA125 and HBME‐1 expression
Authors: BATEMAN A.C. , AL‐TALIB R.K. , NEWMAN T. , WILLIAMS J.H. , HERBERT A.
Published: November 2003

Histological diagnosis of malignant mesothelioma and differentiation from adenocarcinoma is often difficult. Definitive pathological confirmation of malignant mesothelioma requires demonstration of an appropriate immunohistochemical phenotype. Selection of an optimum panel of immunohistochemical antibodies for the reliable identification of malignant mesothelioma is hindered by the absence of a specific immunohistochemical label for mesothelioma cells. Recently, we have found that the ovarian carcinoma cell antibody CA125 labels malignant mesothelioma cells, and the antibody HBME‐1 has been developed as a sensitive mesothelial cell marker. We have compared the immunohistochemical staining patterns achieved with CA125 and HBME‐1 to those obtained using a panel of eight further antibodies in 17 malignant mesotheliomas and 14 primary and secondary adenocarcinomas within lung and pleura. CA125 labelled malignant mesothelioma cells in 15 of 17 cases (88%), and adenocarcinoma cells in seven of 14 cases (50%). HBME‐1 labelled mesothelioma cells in all 17 cases (100%) but also labelled adenocarcinoma cells in 10 of 14 cases (71%). BerEP4 positively labelled one malignant mesothelioma but was negative in the remaining 16 cases and positively labelled nine of 14 adenocarcinomas (64%). Monoclonal anti‐CEA, AUA‐1, CA19.9 and LeuM1 labelled no malignant mesotheliomas and were positive in 10 (71%), nine (64%), eight (57%) and six (43%) of 14 cases of adenocarcinoma, respectively. Diastase‐PAS staining detected neutral mucin in none of the malignant mesotheliomas but in 10 (71%) of the 14 adenocarcinomas. We conclude that CA125 and HBME‐1 do not label mesothelial cells with sufficient specificity to be useful for differentiating malignant mesothelioma from adenocarcinoma, although negative staining with HBME‐1 makes a diagnosis of malignant mesothelioma unlikely. As there remains an absence of a specific positive mesothelial cell marker this distinction is still most reliably made using a panel of antibodies including at least two of the following: anti‐CEA, AUA‐1, BerEP4, LeuM1 and CA19.9, in combination with histochemical assessment of neutral mucin production.
Immunohistochemical assessment of proliferative activity in mammary adenomyoepithelioma
Authors: KOYAMA M. , KUROTAKI H. , YAGIHASHI N. , AIZAWA S. , SUGAI M. , KAMATA Y. , OYAMA T. , YAGIHASHI S.
Published: November 2003

Aims: Two cases of adenomyoepithelioma of the breast were examined by immunohistochemistry to evaluate proliferative activity of epithelial and myoepithelial components. Methods and results: The tumours showed a bicellular pattern of gland‐forming epithelial cells and proliferative myoepithelial cells with clear cytoplasm. They showed foci of monotonous growth of myoepithelial cells devoid of glands with low mitotic rate (1 ∼ 2/10 high‐power fields) and mild cytological atypia. Immunohistochemically, the glandular cells were positive for epithelial membrane antigen, cytokeratin (KL‐1 and CAM5.2) and carcinoembryonic antigen, whereas tumour cells with clear cytoplasm were reactive with muscle‐specific actin (MSA), alpha smooth muscle actin, vimentin, and S100 protein but negative for desmin. Proliferative activities assessed by MIB‐1(Ki‐67)/MSA positive cell index were greater in myoepithelial cells in both cases (19.2% and 17.7%) as compared to those in epithelial cells (MIB‐1/CAM5.2 index: 10.2% and 9.5%). Conclusions: These results might account for the previous findings that myoepithelial components predominate over the epithelial ones in an advanced stage of this tumour as well as in recurrent or metastatic lesions.
Anti‐cytokeratin 5/6: a positive marker for epithelioid mesothelioma
Authors: CLOVER J. , OATES J. , EDWARDS C.
Published: November 2003

Aims: Previous studies using frozen material have suggested that cytokeratin 5 is expressed by pleural mesothelioma but not by adenocarcinoma. In the present study, reactions for cytokeratins 5 and 6 were investigated in 33 pleural mesotheliomas and 27 secondary adenocarcinomas of the pleura using formalin‐fixed, paraffin‐embedded material and a commercially available antibody (anti‐cytokeratin 5/6). Methods and results: All of the adenocarcinomas originated in the peripheral lung. The epithelioid component of the mesotheliomas gave a strongly positive reaction; the reaction in sarcomatoid or desmoplastic areas was absent or weak. Twenty‐two of the adenocarcinomas were negative, in four there was a weak, equivocal reaction, and in one there was patchy positivity. Conclusions: We conclude that this antibody is potentially a useful positive marker for the identification of the epithelioid variant of mesothelioma in formalin‐fixed and paraffin‐embedded material.
Matrix metalloproteinases and tissue inhibitors of metalloproteinases in thyroid C‐cells and medullary thyroid carcinomas
Authors: TOMITA T.
Published: November 2003

Aims: Hyperplastic C‐cells of the thyroid and medullary thyroid carcinomas (MTCs) were studied immunocytochemically for calcitonin, carcinoembryonic antigen (CEA), chromogranin A, matrix metalloproteinase (MMP)‐2 and ‐9, and tissue inhibitor of metalloproteinase (TIMP)‐1 and ‐2. Methods and results: Non‐neoplastic C‐cells were positive for all these substances whereas MTC tumour cells were relatively weaker stained, and thyroid follicular cells were negative for all the substances studied. We have recently reported that pancreatic islet cells and islet cell tumours were positive for MMPs and TIMPs, and, in addition, anterior pituitary cells and pituitary adenomas and parathyroid gland and its adenomas were also positively stained. The MMP‐ and TIMP‐positive endocrine cells correspond to Pearse's APUD cells, derived from neural crest and endodermal cells. Conclusions: MMPs and TIMPs may well be added as newly recognized markers for neuroendocrine cells. The possible function of MMP‐TIMP homeostasis in C‐cells and MTCs is discussed.
Expression and prognostic value of c‐erbB‐2 oncogene product in human phaeochromocytomas
Authors: CASTILLA‐GUERRA L. , MORENO A.M. , FERNÁNDEZ‐MORENO M.C. , UTRILLA J.C. , FERNÁNDEZ E. , GALERA‐DAVISON H.
Published: November 2003

Aims: Amplification of c‐erbB‐2 proto‐oncogene has been reported in endocrine tumours, but the results were unclear and no predictive prognostic value has been established in the case of phaeochromocytoma. We investigated the immunohistochemical expression of c‐erbB‐2 oncogene in 34 cases of human phaeochromocytoma (27 sporadic, seven familial type MEN (multiple endocrine neoplasm)) in order to find out if it could be used to differentiate sporadic and familial forms and whether c‐erbB‐2 expression is related to tumour biological behaviour. Methods and results: All the cases showed diffuse, generally heterogeneous, intracytoplasmic granular c‐erbB‐2 staining. The percentage of tumour cells expressing c‐erbB‐2 was used as the comparative variable. The percentage of c‐erbB‐2 positive cells had a statistically significant (P < 0.001) relationship with tumour aggressiveness, as manifested by the presence of distant metastasis or association with other malignant neoplasms. We also found significantly higher levels (P = 0.007) of c‐erbB‐2 overexpression in MEN phaeochromocytoma than in sporadic cases. Conclusions: These results clarify the important role of c‐erbB‐2 proto‐oncogene in the pathogenesis of human phaeochromocytoma and confirm the unfavourable prognostic significance of c‐erbB‐2 expression.
Umbilical cord ‘pseudo‐vasculitis’ following second trimester fetal death: a clinicopathological and immunohistochemical study of 13 cases
Authors: GENEST D.R. , GRANTER S. , PINKUS G.S.
Published: November 2003

Amniotic fluid bacterial infection is an occasional cause of second trimester septic abortion. We describe an autolysis‐related histological artifact, umbilical cord ‘pseudo‐vasculitis’, which can erroneously implicate amniotic bacterial infection in fetal death. Clinicopathological features of 13 second trimester fetal deaths with umbilical cord pseudo‐vasculitis are reported. In four cases (31%), an incorrect pathological diagnosis of umbilical vasculitis had initially been rendered. Umbilical cords from five cases of pseudo‐vasculitis and one comparison fetus (18‐week septic abortion with true umbilical vasculitis), were studied with chloroacetate esterase and with immunohistochemical staining for myeloperoxidase, muscle‐specific actin (HHF35) and smooth muscle actin. Histologically, umbilical pseudo‐vasculitis exhibited numerous small, rounded, degenerating cells with irregular, multilobed nuclei (closely resembling neutrophils) located within the umbilical vessel wall. Immunohistochemical studies demonstrated that all cells resembling neutrophils were of smooth muscle origin. Moderate to severe fetal autolysis was present in all cases of umbilical pseudo‐vasculitis, suggesting that this finding represents autolysis of umbilical vascular smooth muscle secondary to post mortem fetal retention. Vascular smooth muscle in other fetal and placental locations did not demonstrate the finding, suggesting that this striking degenerative artifact of smooth muscle is restricted to the umbilical cord.
Calcifying fibrous pseudotumour of visceral peritoneum
Authors: KOCOVA L. , MICHAL M. , SULC M. , ZAMECNIK M.
Published: November 2003

Aims: Calcifying fibrous pseudotumour, a recently characterized rare but distinctive lesion, has been reported previously only in soft tissues and in the pleura. We describe four cases of calcifying fibrous pseudotumour which arose in the visceral peritoneum of the small bowel and stomach. Methods and results: In contrast to the calcifying fibrous pseudotumour of soft tissues which generally occurs in children, all four cases in our report occurred in adults. We consider the calcifying fibrous pseudotumour to be a distinctive tissue reaction. Conclusions: Its characteristic appearance of concentric whorls of lamellated collagen associated with psammoma bodies and other forms of dystrophic calcification is entirely different from all other lesions displaying hyaline collagenous deposits, such as inflammatory pseudotumour, desmoid, hyalinizing granuloma or fibrous plaques of peritoneum.
Diagnostic criteria for pseudomalignant osteoblastoma
Authors: CHEUNG F.M.F. , WU W.C. , LAM C.K. , FU Y.K.
Published: November 2003

Aims: Osteoblastoma is a rare bone tumour and differentiation from osteosarcoma is occasionally a diagnostic problem. The difficulty is further compounded when such a lesion microscopically shows cytological or architectural atypia. Methods and results: A case of pseudomalignant osteoblastoma occurring in the left iliac crest of a 34‐year‐old Chinese woman is presented. Radiographic findings were non‐diagnostic, but favoured a benign yet locally aggressive lesion. Histologically the diagnosis was based on the findings of classical osteoblastoma areas harbouring pseudomalignant osteoblasts in the absence of mitoses, but there were co‐existing atypical architectural features also. Conclusion: The significance and pathogenesis of the ‘permeative pattern’ are discussed. Difficulties in differentiating borderline lesions are emphasized.
Production and characterization of a new monoclonal antibody effective in recognizing the CD3 T‐cell associated antigen in formalin‐fixed embedded tissue
Authors: STEWARD M. , BISHOP R. , PIGGOTT N.H. , MILTON I.D. , ANGUS B. , HORNE C.H.W.
Published: November 2003

Phenotypic analysis of lymphoproliferative disorders is now considered mandatory for accurate classification which is the basis for optimum patient management. This is presently carried out in most cases using a range of antibodies recognizing B and T‐cell antigens effective in paraffin sections, and an antibody to CD3 is currently a key member of such panels, indicating T‐cell phenotype. Current antibodies to CD3 are polyclonal with the inherent disadvantages of this type of reagent compared to monoclonal antibodies. In this study, we have used a recombinant fusion protein representing part of the epsilon subunit of the CD3 molecule to generate a novel monoclonal antibody (NCL‐CD3‐PS1) effective in paraffin sections. The antibody has been characterized biochemically and by immunohistochemistry using a wide range of normal and pathological tissues. Lineage and phenotype specificity have been supported in our study and results from other laboratories are awaited with interest.
Impact of donor MHC class I or class II antigen deficiency on first‐ and second‐set rejection of mouse heart or liver allografts
Authors: QIAN S. , FU F. , LI Y. , LU L. , RAO A. S. , STARZL T. E. , THOMSON A. W. , FUNG J. J.
Published: November 2003

The influence of donor major histocompatibility complex (MHC) class I‐ or class II‐deficiency on the initiation of first‐ and second‐set rejection of mouse heart and liver allografts was examined. C3H (H‐2k) mice received heterotopic cardiac or orthotopic liver grafts from unmodified B10 (H‐2b), B6 (H‐2b), b2m (H‐2b; class I deficient) or AB0 (H‐2b; class II deficient) donors. Organ survival was also investigated in C3H recipients that had been presensitized by a normal B10 skin graft 2–3 weeks before heart or liver transplantation. The absence of cell surface MHC class I or class II resulted in significant prolongation of primary cardiac allograft survival. Three of seven (43%) MHC class I‐deficient, and two of five (40%) class II‐deficient heart grafts were accepted indefinitely (survival time >100 days). Thus both MHC class I and class II molecules appear to be important for the elicitation of first‐set rejection in the heart allograft model. All liver allografts survived >100 days in normal recipients. In C3H recipients that had been presensitized by a B10 skin graft, however, both heart and liver grafts from AB0 (class II deficient) donors underwent accelerated rejection (median survival time [MST] 3 and 4 days, respectively). In contrast, liver grafts from class I‐deficient mice (b2m) were still accepted indefinitely by B10 skin‐presensitized C3H recipients, whereas class I‐deficient hearts survived significantly longer than those from class II‐deficient or normal donors. These data demonstrate that the expression of donor MHC class I, and not class II is crucial in initiating second‐set organ allograft rejection. In vitro monitoring revealed that at the time of organ transplant, both splenocytes and serum of the skin‐presensitized animals displayed high cytotoxicity against AB0 (class II‐deficient) but not against b2m (class I‐deficient) targets.
Ability of spleen, peritoneal cavity, and lymph node B cells to reconstitute serum immunoglobulin in SCID mice
Authors: RIGGS J. , STOWERS R.
Published: November 2003

The impact of intrinsic B lymphocyte heterogeneity and of microenvironmental influences on serum immunoglobulin production by B cells was examined by intravenous (i.v.) and intraperitoneal (i.p.) transfer of BALB/c and BALB.xid (X‐chromosome‐linked immune‐defective; XID) lymph node (LN), splenic (SP) and peritoneal cavity (PerC) cells into severe‐combined immune‐defective (SCID) mice. The results indicate that each B‐cell source restores all immunoglobulin classes within 5 weeks of transfer, the rates for each isotype, however, differ between the B‐cell sources. Serum IgM levels were restored most rapidly by PerC cell transfer, followed by SP and LN cell transfer. In addition, normal immunoglobulin levels were reached in the absence of complete lymphoid reconstitution. Serum immunoglobulin phenotypes characteristic of the donor strain, e.g. reduced IgM and IgG3 production by XID B cells, were maintained after transfer into the SCID recipient. Microenvironmental influences were indicated by reduced immunoglobulin production after i.p. transfer and after i.v. transfer into irradiated SCID recipients. The data show that both B‐cell type and microenvironment play significant roles in generating the heterogeneous pool of B cells required for humoral immunity.
Expression and function of membrane regulators of complement on rat astrocytes in culture
Authors: ROGERS C. A. , GASQUE P. , PIDDLESDEN S. J. , OKADA N. , HOLERS V. M. , MORGAN B. P.
Published: November 2003

Human astrocytes express CD59, decay accelerating factor and membrane cofactor protein to restrict the damaging effect of complement (C) activation on their cell surface. 5I2 antigen (5I2 Ag) is the functional analogue of the latter two proteins in rats. We here demonstrate the surface expression on rat astrocytes of CD59 and 5I2 Ag and use sodium dodecyl sulphate–polyacrylamide gel electrophoresis and Western blotting to confirm their identity and to quantify expression. Rat CD59 (MW 20 000) was expressed at 720 × 103 molecules per cell and 5I2 Ag (MW 58 000 and 64 000) at 625 × 103 molecules per cell. Reverse transcription–polymerase chain reaction using specific oligonucleotide primers demonstrated expression of mRNA for each protein. Twenty‐four‐hour stimulation with inflammatory cytokines (interferon‐γ, tumour necrosis factor‐α, interleukins‐1β, ‐2 and ‐6) or phorbol myristate acetate had no significant effect on the level of expression of either protein as determined by Western blotting. Lysis caused by classical pathway activation of C in human or rat serum was enhanced by blocking the function of CD59 and 5I2 Ag on rat astrocytes with monoclonal antibodies.
Fetal lambs are depleted of IgM+ cells following a single injection of an anti‐IgM antibody early in gestation
Authors: PRESS C. MCL. , REYNOLDS J. D. , MCCLURE S. J. , SIMPSON‐MORGAN M. W. , LANDSVERK T.
Published: November 2003

B‐cell depleted fetal sheep were created following a single injection of an anti‐IgM monoclonal antibody early in gestation. Six sheep fetuses were given a single intraperitoneal injection of a monoclonal antibody directed against IgM at 63 days of gestation (gestation in sheep = 150 days). The fetuses were killed at 138–142 days of gestation and lymphoid tissues were collected for subsequent light microscopy and immunohistochemical examination. The ileal and jejunal Peyer's patch (PP) follicles in four of the six injected fetuses were markedly reduced in size. Cells in the rudimentary follicles of the ileal PP of these animals showed no reactivity for IgM and most were negative for CD45. The dome regions contained many T cells, which were predominantly CD8+ cells and included γδ T cells. The interfollicular areas of the PP of the markedly affected fetuses contained large populations of T cells. The spleen and lymph nodes were also markedly depleted of IgM+ cells and these tissues contained only a small, scattered population of weakly IgM+ cells. Follicular accumulations of IgM+ cells were absent. Large populations of T cells were present in the white pulp of the spleen and cortex of the lymph nodes. The liver did not contain IgM+ cells and the medulla of the thymus was depleted of IgM+ cells. The results of this study suggest that a surface IgM+ B‐cell population is present in the sheep fetus at 63 days of gestation, which is essential for the colonization of the ileal PP and subsequent B‐cell development.
Late‐term fetal thymectomy does not prevent the development of gut‐homing T cells after birth
Authors: CAHILL R. N. P. , KIMPTON W. G. , WASHINGTON E. A. , DUDLER L. , TRNKA Z.
Published: November 2003

Tissue‐specific circulation of T cells is a critical element in the integration of systemic immune responses. Current models of T‐cell migration suggest that homing specificities of T cells for tissues such as gut and skin are generated outside the thymus as a result of activation of virgin T cells by antigen in lymph nodes. We have used the sheep fetus (which is immunologically virgin and contains no memory or effector T‐cell subsets) to examine the migration of 51Cr‐labelled T cells in vivo. We report that gut‐homing T cells are not present in the fetus and that gut‐homing T cells from postnatal lambs home normally to fetal gut. Fetal thymectomy performed immediately prior to birth failed to prevent the development of gut‐homing T cells in postnatal life. Gut‐homing specificities on T cells are thus acquired extrathymically.
Repopulation of blood lymphocyte sub‐populations in rheumatoid arthritis patients treated with the depleting humanized monoclonal antibody, CAMPATH‐1H
Authors: BRETT S. , BAXTER G. , COOPER H. , JOHNSTON J. M. , TITE J. , RAPSON N.
Published: November 2003

Patients with severe rheumatoid arthritis who had failed treatment with conventional therapies were treated with a course of five or 10 daily intravenous infusions of CAMPATH‐1H, a humanized antibody against the CD52 antigen, resulting in profound depletion of peripheral blood mononuclear cells. During the subsequent 18 months, lymphocytes were analysed for sub‐populations by fluorescence‐activated cell sorter (FACS) and for proliferation in response to polyclonal T‐cell stimulation with anti‐CD3 or staphylococcal enterotoxin B (SEB). Treatment resulted in almost complete depletion of lymphocytes from the blood followed by gradual repopulation. CD16+ natural killer (NK) cells and CD14+ monocytes returned to pretreatment levels within 1–2 months. CD19+ B cells returned to within 50% of pre‐treatment levels by day 66 and to within normal range by day 150, whereas CD8+ T cells recovered to 50% of pretreatment levels by day 66, but did not show any further increase during the rest of the study period. The most profound effects were on the CD4+ T lymphocyte sub‐population, as the mean CD4+ count did not increase above 20% of pre‐treatment level at any time during the study period (550 days), at all the doses tested. The T cells which initially repopulated the blood 1–2 months after treatment, nearly all expressed the activation markers human leucocyte antigen (HLA)‐DR and CD45RO, although the percentage of T cells expressing these molecules gradually declined to normal levels over time. Proliferative responses to polyclonal T‐cell stimulation (anti‐CD3 and SEB) were also significantly reduced in the first few months after treatment, but recovered to pre‐treatment levels by day 250. The relationship between these observations and the clinical response is discussed.
The Epstein–Barr virus‐binding site on CD21 is involved in CD23 binding and interleukin‐4‐induced IgE and IgG4 production by human B cells
Authors: HENCHOZ‐LECOANET S. , JEANNIN P. , AUBRY J.‐P. , GRABER P. , BRADSHAW C. G. , POCHON S. , BONNEFOY J.‐Y.
Published: November 2003

Human CD21 has previously been described as a receptor for the C3d,g and iC3b proteins of complement, as a receptor for the gp350/220 envelope glycoprotein of the Epstein–Barr virus (EBV) and also as a receptor for interferon‐α (IFN‐α). Structurally, CD21 consists of 15 to 16 short consensus repeats (SCR) of 60 to 75 amino acids followed by a transmembrane domain and an intracytoplasmic region. We reported that CD23, a low‐affinity receptor for IgE (FcεR2), is a new functional ligand for CD21. We recently found that the sites of interaction of CD23 on CD21 are on SCR 5 to 8 and 1–2. The first site is a lectin–sugar type of interaction and the second site is a protein–protein interaction. We report here that amongst the other ligands for CD21 (EBV, C3d,g and IFN‐α), only EBV is able to inhibit the binding of CD23 to CD21. Furthermore, even a peptide from gp350/220 of EBV known to bind to CD21 is able to decrease CD23 binding to CD21. Since CD23/CD21 pairing is important in the control of IgE production, we tested the effect of the EBV‐derived peptide on immunoglobulin production from peripheral blood mononuclear cells and purified tonsillar B cells. Interestingly, the EBV‐peptide inhibited IgE and IgG4 production induced by interleukin‐4, in a dose‐dependent manner. The same results were obtained using either peripheral blood mononuclear cells or purified tonsillar B cells. Another CD21 ligand, C3, did not affect binding of CD23 to CD21 nor the production of IgE and IgG4. This study indicates that blocking CD23 binding to CD21 SCR 2 on human B cells selectively modulates immunoglobulin production.
The role of tumour necrosis factor‐α in combination with interferon‐γ or interleukin‐1 in the induction of immunosuppressive macrophages because of Mycobacterium avium complex infection
Authors: TOMIOKA H. , MAW W. W. , SATO K. , SAITO H.
Published: November 2003

The role of some cytokines including tumour necrosis factor‐α (TNF‐α), interleukin‐1α (IL‐1α), interferon‐γ (IFN‐γ), transforming growth factor‐β (TGF‐β) and interleukin‐6 (IL‐6) in the generation of immunosuppressive macrophages (MΦs) in host spleen cells of Mycobacterium avium complex (MAC)‐infected mice was studied. MΦ populations with potent suppressor activity against concanavalin A (Con A)‐induced mitogenesis of splenocytes (SPCs) were elicited not only in euthymic but also in athymic nude mice during MAC infection. The suppressor MΦs are, therefore, inducible not only through a T‐cell‐dependent mechanism but also through T‐cell‐independent mechanism. However, MAC‐induced MΦs of athymic mice displayed about four times lower suppressor activity than those of euthymic mice, indicating that mature T cells are important for MΦ activation to the highly immunosuppressive state. Anti‐TNF, anti‐IFN‐γ, and anti‐TGF‐β antibodies (Abs) but not anti‐IL‐6 Ab inhibited in vivo generation of MAC‐induced immunosuppressive MΦs, and the neutralizing efficacy was in the order of anti‐IFN‐γ Ab > anti‐TNF Ab > anti‐TGF‐β Ab. The effects of TNF‐α, IL‐1α, IL‐6, and IFN‐γ alone or combinations of them upon the acquisition of the suppressor activity by cultured splenic MΦs were studied. When normal splenic MΦs were treated with each cytokine for 3 days, TNF‐α, IFN‐γ, and IL‐1α alone caused a slight elevation of their suppressive activity. Treatment of the normal MΦs with the combination of either TNF‐α + IL‐1α or TNF‐α + IFN‐γ yielded a marked increase in the suppressor activity, followed by IL‐1α + IFN‐γ. These findings indicate the imortant roles of TNF‐α, IFN‐γ, and IL‐1α in the generation of MAC‐induced suppressor MΦs.
Secretion of cytokines by natural killer cells primed with interleukin‐2 and stimulated with different lipoproteins
Authors: DE SANCTIS J. B. , BLANCA I. , BIANCO N. E.
Published: November 2003

Natural killer (NK) cells were shown to secrete differentially interleukins (IL), IL‐1α, IL‐1β, IL‐2, IL‐8, interferon‐γ (IFN‐γ), tumour necrosis factor‐α (TNF‐α), granulocyte–macrophage colony‐stimulating factor (GM‐CSF), and leukaemia inhibitory factor (LIF) upon stimulation with optimal concentrations of chylomicrons (CM), very‐low‐density lipoprotein (VLDL), low‐density lipoprotein (LDL), high‐density lipoprotein (HDL) or acetyl‐modified low‐density lipoprotein (AcLDL). CM, VLDL, LDL and AcLDL induced LIF secretion which was absent in non‐stimulated cells. CM, VLDL, and LDL did not affect IL‐1α secretion. CM stimulated IL‐8>TNF‐α>IL‐1α>IL‐2=IFN‐γ, and decreased seventeenfold GM‐CSF secretion. VLDL stimulated IL‐8 secretion>IL‐1α=IL‐2>IFN‐γ>TNF‐α and decreased fivefold GM‐CSF secretion. LDL stimulated IL‐8 secretion>IL‐1α>IL‐2=IFN‐γ, it did not modify TNF‐α and inhibited five hundred‐fold GM‐CSF secretion. HDL stimulated IL‐2 secretion=IFN‐γ>IL‐8, it decreased GM‐CSF secretion>IL‐1α>IL‐1β>TNF‐α without affecting LIF. AcLDL stimulated IL‐8 secretion >TNF‐α>IL‐1α>IL‐2=IFN‐γ=IL‐1β, and decreased GM‐CSF secretion eightfold. When NK cells were primed with 10, 100 or 500 IU/ml of IL‐2 before the addition of lipoproteins, a decrease in the secretion of cytokines was observed as compared with cells primed with IL‐2 only. Differences in cytokine secretion were observed among the diverse type of lipoproteins used for cell stimulus. Thus, lipoproteins may condition NK cytokine secretion and cell activation.
Alleviation of insulitis in NOD mice is associated with expression of transgenic MHC E molecules on primary antigen‐presenting cells
Authors: PILSTRÖM B. , BÖHME J.
Published: November 2003

Major histocompatability complex (MHC) class II genes are important in the pathogenesis of insulin‐dependent diabetes mellitus (IDDM) both in the mouse and in man. The non‐obese diabetic (NOD) mouse, which is a good model for human IDDM, has a particular MHC class II with an A complex consisting of Aαd and the unique Aβg7 chain, as well as an absent E molecule due to a deletion in the Ea promoter region. Transgenic insertion of a functional Ea gene protects against insulitis and diabetes, but when the transgene expression is restricted to certain compartments of the immune system by deleting parts of the promoter region, the protection against insulitis is disrupted. We have analysed three promoter‐mutated lines where one lacks expression on B cells and has a reduced expression on approximately 1/3 of the dendritic cells and macrophages (Sma), one lacks thymic cortical expression and has a slightly reduced B‐cell expression (ΔX), and one lacks expression in the thymic medulla, on macrophages, dendritic cells and about half of the B cells (ΔY). None of these lines is protected against insulitis, but Sma and ΔX display a reduced intensity of insulitis, with an average of 10–15% of the islets infiltrated in each mouse, while ΔY resembles non‐transgenic mice with 30–35% infiltrated islets. Bone‐marrow chimeras between Sma and ΔY mice demonstrate that peripheral cells of Sma origin reduce insulitis significantly when developed in the ΔY host, while insulitis is enhanced when ΔY bone marrow is given to Sma mice. This shows that E expression on the primary antigen‐presenting macrophages and dendritic cells is of crucial importance to the alleviation of insulitis.
Expression of rat CD59: functional analysis confirms lack of species selectivity and reveals that glycosylation is not required for function
Authors: RUSHMERE N. K. , TOMLINSON S. , MORGAN B. P.
Published: November 2003

This study reports the expression and functional characterization of the rat analogue of the human complement regulatory molecule CD59. We here decribe the expression in chinese hamster ovary (CHO) cells of rat CD59 and a modified rat CD59 in which an N‐glycosylation site at Asn‐16 has been deleted by point mutation. The complement‐inhibiting capacity of these two forms of rat CD59 has been analysed and compared. Expressed rat CD59 efficiently inhibited complement lysis of CHO cells when rat serum was used as a source of complement and also inhibited lysis by complement from all other species tested, confirming that rat CD59 displayed little or no species restriction of activity. Blocking of expressed rat CD59 with a monoclonal antibody abrogated the inhibition of lysis for all sources of complement, confirming that the expressed molecule was responsible for the protection. The glycosylation mutant had a much reduced molecular weight on sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS–PAGE) (12 000 MW as compared with 20 000–28 000 MW for unmutated), confirming that it was unglycosylated. However, the glycosylation mutant had complement‐inhibitory activity which was at least as potent as that of the unmutated molecule, demonstrating that the large, N‐linked carbohydrate moiety was not required for function.
Interferons α/β inhibit IL‐7‐induced proliferation of CD4− CD8− CD3− CD44+ CD25+ thymocytes, but do not inhibit that of CD4− CD8− CD3− CD44− CD25− thymocytes
Authors: SU D.‐M. , WANG J. , LIN Q. , COOPER M. D. , WATANABE T.
Published: November 2003

Type 1 interferons (IFN‐α/β) have recently been shown to inhibit interleukin‐7 (IL‐7)‐induced growth and survival of early B‐lineage cells. The CD3− CD4− CD8− (triple negative; TN) thymocytes from normal mice strongly proliferated upon stimulation with IL‐7 in suspension culture. Such an IL‐7‐induced proliferation was suppressed by the addition of IFN‐α/β, but a fraction of the TN thymocytes still showed proliferation. The IL‐7‐induced growth of TN thymocytes from scid mice, which lack the CD44− CD25− subpopulation, was completely inhibited by the addition of IFN‐α/β. The IL‐7 induced proliferation of CD4− CD8− thymocytes from T‐cell receptor (TCR) transgenic mice, the majority of which are CD3+ CD44− CD25−, was resistant to IFN‐α/β‐mediated suppression. In fetal thymus organ cultures (FTOC), the addition of IL‐7 greatly increased the population of CD4− CD8− CD44+ CD25+ thymocytes and IFN‐α/β inhibited this IL‐7‐driven expansion. In contrast, the addition of IL‐7 markedly decreased the percentages of CD4− CD8− CD3− CD44− CD25− cells, and IFN‐α/β reversed the effect and increased the subpopulations of CD44− CD25+ and CD44− CD25.− Finally, IFN‐β mRNA was found to be expressed in the thymus. The data suggest that type 1 interferons inhibit IL‐7‐driven proliferation of TN thymocytes, but do not block the normal differentiation process.
Evidence for a reduced chemokine response in the lungs of beige mice infected with Mycobacterium avium
Authors: FLORIDO M. , APPELBERG R. , ORME I. M. , COOPER A. M.
Published: November 2003

The basis of the increased susceptibility of beige mice to Mycobacterium avium infections is still not clearly understood. In this study we examined the growth of three virulent strains of M. avium in beige mice and normal C57BL/6 controls. Depletion of natural killer (NK) cells by administration of anti‐asialo GM1 antisera did not affect the growth of M. avium in any of the groups of animals. Similarly, interferon‐γ (IFN‐γ) gene‐disrupted mice were more susceptible to infection than control mice but the growth of M. avium was not further affected by NK‐cell depletion. In terms of effector immunity, beige mice showed enhanced expression of IFN‐γ and tumour necrosis factor‐α (TNF‐α) when compared with wild‐type C57BL/6 mice. In agreement with these results, I‐A and interferon‐inducible protein (IP‐10) expression was also higher in beige mice than in wild‐type animals, as was expression of the chemokines macrophage inflamatory protein‐2 (MIP‐2) and macrophage chemotactic protein (MCP‐1) during latter stages of the infection. However, over the first few weeks of the infection, when the susceptibility of the beige mouse lung first becomes evident, MIP‐1β and MIP‐2 chemokine expression in the lungs was lower in beige mice than in wild‐type animals. These data indicate, therefore, that the increased susceptibility of beige mice to M. avium infection in the lung is not due to lack of NK‐cell activity, nor can it be explained in terms of the effector cytokine response. Instead, the lower early expression of the neutrophil chemoattractants MIP‐1β and MIP‐2 in the lungs of beige mice tends to suggest that the enhanced susceptibility of these mice to M. avium infection may be due in part to defective recruitment of neutrophils or other cells responsive to these specific chemokines.
Characterization of Marek’s disease herpesvirus‐specific cytotoxic T lymphocytes in chickens inoculated with a non‐oncogenic vaccine strain of MDV
Authors: OMAR A. R. , SCHAT K. A.
Published: November 2003

Previously we have reported that reticuloendotheliosis virus (REV)‐transformed cell lines expressing Marek’s disease virus (MDV) genes pp38, meq or gB were lysed by syngeneic MDV‐specific splenocytes from major histocompatibility complex (MHC):B9B19 and MHC:B1B21 chickens. In contrast, REV‐transformed cell lines expressing the MDV gene ICP4 were only lysed by syngeneic MDV‐specific splenocytes from MHC:B21B21 chickens. In this study we report that this syngeneic cell‐mediated immune response is induced by cytotoxic T lymphocytes (CTL). Splenocytes from MDV vaccine strain, SB‐1 inoculated MHC:B19B19 and MHC:B21B21 chickens, depleted for CD4+, CD8+, TCRγδ+, TCRαβ1+ and/or TCRαβ2+ cells, were used as effector cells in chromium‐release assays. Effector cells depleted of CD8+ or TCRαβ1,+ but not TCRγδ+ or TCRαβ2+, markedly reduced the MDV‐specific release. Depletion of CD4+ effector cells did not influence the specific release significantly. This is the first report on identification of virus‐specific CD8+ CTL in chickens inoculated with a non‐oncogenic vaccine strain of MDV.
Scanning electron microscope appearances of Lightspeed instruments used clinically: a pilot study
Authors: Marending M. , Lutz F. , Barbakow F.
Published: November 2003

The cutting heads of the first 18 instruments in six sets of clinically used Lightspeed instruments (sizes 20–65) were examined in a scanning electron microscope (SEM). Three sets had been used by full‐time faculty and discarded as follows: instrument sizes 20–32.5 after 18 treated root canals; instrument sizes 35–47.5 after 36 treated root canals; and instrument sizes 50–65 after 54 treated root canals. Three other sets had been used by a private clinician who discarded all Lightspeed instruments after preparing 20 root canals. The cutting heads of the 108 instruments were examined for the presence of surface debris, metal strips, metal flash, disrupted cutting edges, microfractures, tears, pitting and fretting and fatigue cracks. Fewer instruments ultrasonically cleaned before examination in the SEM had debris than those not cleaned before examination. The prevalence of metal strips and metal flash was similar for both groups. In contrast, microfractures and tears were more prevalent in instruments used by the full‐time faculty group, who used their instruments more frequently before discarding them. Pitting and fretting occurred equally frequently in both groups whilst no fatigue cracks were noted in any of the 108 instruments. Overusage of Lightspeed instruments may predispose the flutes of the cutting heads to microfractures and clinicians should be aware of this possibility.
The observation of intact hepatic endothelial cells by cryo‐electron microscopy
Authors: Braet F. , Bomans P. H. H. , Wisse E. , Frederik P. M.
Published: November 2003

Liver sinusoidal endothelial cells (LSECs) can optimally be imaged by whole mount transmission electron microscopy (TEM). However, TEM allows only investigation of vacuum‐resistant specimens and this usually implies the study of chemically fixed and dried specimens. Cryo‐electron microscopy (cryo‐EM) can be used as a good alternative for imaging samples as whole mounts. Cryo‐EM offers the opportunity to study intact, living cells while avoiding fixation, dehydration and drying, at the same time preserving all solubles and water as vitrified ice. Therefore, we compared the different results obtained when LSECs were vitrified using different vitrification conditions. We collected evidence that manual blotting at ambient conditions and vitrification by the guided drop method results in the production of artefacts in LSECs, such as the loss of fenestrae, formation of gaps and lack of structural details in the cytoplasm. We attribute these artefacts to temperature and osmotic effects during sample preparation just prior to vitrification. By contrast, by using an environmentally controlled glove box and a vitrification robot (37 °C and 100% relative humidity), these specific structural artefacts were nearly absent, illustrating the importance of controlled sample preparation. Moreover, data on glutaraldehyde‐fixed cells and obtained by using different vitrification methods suggested that chemical prefixation is not essential when vitrification is performed under controlled conditions. Conditioned vitrification therefore equals chemical fixation in preserving and imaging cellular fine structure. Unfixed, vitrified LSECs show fenestrae and fenestrae‐associated cytoskeleton rings, indicating that these structures are not artefacts resulting from chemical fixation.
A composite method for the determination of the chirality of single walled carbon nanotubes
Authors: Meyer R. R. , Friedrichs S. , Kirkland A. I. , Sloan J. , Hutchison J. L. , Green M. L. H.
Published: November 2003

An approach to the unambiguous determination of the conformation of individual single walled nanotubes utilizing high‐resolution transmission electron microscopy and digital image processing is described. The exit plane wave of single walled nanotubes restored from a focal series of images is used in a stepwise characterization procedure utilizing both the phase of the real space restoration and its Fourier transform. A combination of these complementary characterization steps yields an accurate measurement of the chiral vector for an individual nanotube.
Polyclonal T‐cell responses to Plasmodium falciparum gametocytes in malaria nonexposed donors
Authors: GOODIER MARTIN R. , TARGETT GEOFFREY A.T.
Published: November 2003

Human peripheral blood T‐cells mount vigorous proliferative responses to asexual stage parasites of P. falciparum regardless of whether the donor has been exposed to the parasite. Here using highly purified P. falciparum gametocytes we show that the same is also true for this stage of the parasite. Gametocytes, like immature trophozoites, preferentially activate CD4+ T cells. γδ T‐cell activation, commonly observed in response to mature schizonts or supernatants from asexual‐stage cultures, does not occur. Furthermore, the CD4+ T‐cell blasts stimulated by gametocytes show a similar pattern of T‐cell receptor variable region (TCRVβ) usage to those stimulated by asexual parasites and there is no preferential usage of TCRVβ elements. The CD4+ T‐cell precursor frequencies for gametocyte and asexual trophozoite responses are remarkably similar. ‘Cross‐reactivity’ of gametocyte and asexual stage responses was confirmed by selective depletion of cells responding to particular stages of the parasite using the cytostatic drug cytosine arabinoside (Ara‐C). These results suggest that the CD4+ T‐cell responses from malaria nonexposed donors are common to gametocytes and asexual trophozoites.