bluuebunny/crossref_metadata_2025 huggingface dataset:
Distinct δ T cell receptor repertoires in monozygotic twins concordant for coeliac disease
Authors: Holtmeier W , Rowell D L , Nyberg A , Kagnoff M F
Published: November 2003

One of the hallmarks of coeliac disease, both active and treated, is an increased number and proportion of γ/δ intraepithelial T lymphocytes in the small intestinal mucosa, and an increased number of γ/δ T cells in the small intestinal mucosa of coeliac disease patients has been associated with the inheritance of specific HLA class II DQ alleles. Nonetheless, the contribution of genetic factors to the development of the T cell receptor (TCR) δ repertoire in coeliac disease is not known. We have assessed the contribution of genetic factors to development of the TCR δ repertoire in coeliac disease, by characterizing the junctional diversity of TCR δ transcripts expressed in the intestine and peripheral blood of a pair of monozygotic (MZ) twins concordant for coeliac disease. TCR Vδ1, Vδ2 and Vδ3 transcripts from small intestinal and colon biopsies, and from peripheral blood mononuclear cells, were amplified by polymerase chain reaction (PCR) and the complementarity determining region (CDR)3 domains of TCR δ transcripts were analysed by denaturing PAGE and direct nucleotide sequencing. The repertoire of TCR δ transcripts and CDR3 amino acid motifs in the intestine and peripheral blood of MZ twins concordant for coeliac disease exhibited no overlap. The TCR δ repertoire in each twin was oligoclonal, and complexity of the junctional regions of their TCR δ transcripts was typical of the repertoire in healthy adults. Thus, genetically identical individuals with coeliac disease have distinct, non-overlapping TCR δ repertoires. Moreover, genetic factors that determine disease susceptibility do not appear to select for specific TCR δ sequences or CDR3 amino acid motifs.
Cytotoxic effect of autocrine and macrophage-derived nitric oxide on cultured rat mesangial cells
Authors: Hruby Z , Beck K-F
Published: November 2003

Expression of the inducible form of nitric oxide synthase (iNOS) has been found to be up-regulated in cytokine-stimulated mesangial cells (MC) and in experimental glomerulonephritis. Since direct toxicity of nitric oxide (NO) has been implicated in damage of bacteria, neoplastic and intact pancreatic cells, we investigated whether NO is cytotoxic to cultured MC, which may be relevant to pathogenesis of glomerular injury. MC isolated from rat glomeruli generated substantial amounts of nitrite, the stable NO end-product, when cells were stimulated with IL-1β and tumour necrosis factor-alpha (TNF-α). Total DNA synthesis was significantly reduced in the presence of IL-1β and TNF-α, and this effect was completely reversed by NG-monomethyl-l-arginine (l-NMMA), an inhibitor of iNOS. Stimulation of MC with IL-1β and TNF-α caused remarkable toxicity to these cells, measured by the MTT test (3-(4,5-dimethylthiazol-2-yl)2,5-diphenyl tetrazolium bromide cleavage, specific cytotoxicity 41.5 ± 20.3%), and much less prominent MC lysis (3H-thymidine release, specific cytolysis 11.5 ± 5.3%). Toxic effects of cytokines were fully reversible by the iNOS inhibitor. Lipopolysaccharide (LPS) and interferon-gamma (IFN-γ), but not IL-1β and TNF-α, induced rat peritoneal macrophages to produce large amounts of nitrite. In co-culture, such prestimulated macrophages had significantly cytotoxic (MTT test 62.9 ± 19.9%) and cytolytic (3H-thymidine release 57.9 ± 13.8%)effects on MC. Again, this toxicity was totally inhibited in the presence of l-NMMA. We conclude from these results that cytokine-stimulated generation of NO by MC or macrophages is directly toxic to MC, and may play a role in pathogenesis of glomerular injury involving mesangiolysis.
Novel autoantigens of perinuclear anti-neutrophil cytoplasmic antibodies (P-ANCA) in ulcerative colitis: non-histone chromosomal proteins, HMG1 and HMG2
Authors: Sobajima J , Ozaki S , Osakada F , Uesugi H , Shirakawa H , Yoshida M , Nakao K
Published: November 2003

Anti-neutrophil cytoplasmic antibodies (ANCA) in sera from ulcerative colitis (UC) patients have been described as reacting with proteins in the granules of human neutrophils such as cathepsin G and lactoferrin and with yet unidentified antigens. Here we report the existence of a new member of perinuclear ANCA (P-ANCA) in UC patients. In the previous study, we found that UC patients had a novel P-ANCA against neutrophil 28-kD protein. In this study, we purified the same antigens from HL-60 lysates by using reversed phase high-performance liquid chromatography, and revealed that the 28-kD antigen consisted of two different proteins. The N-terminus amino acids of these proteins are identical with those of high mobility group (HMG) non-histone chromosomal proteins HMG1 and HMG2. Immunoblotting analysis of human neutrophil lysates using rabbit anti-HMG1/2 antisera revealed a single band of 28 kD, and the 28-kD band detected by immunoblotting analysis using patient's serum IgG completely disappeared after preincubation with a mixture of HMG1 and HMG2. Furthermore, rabbit anti-HMG1/2 antisera showed a perinuclear staining pattern in indirect immunofluorescence studies using ethanol-fixed neutrophils. These data demonstrate that HMG1 and HMG2 are novel target antigens of P-ANCA. HMG1 and HMG2 are distributed in the nuclei and cytoplasm of eukaryotic cells and act as transcription factors. Their intracellular localization and functions are distinct from those of the previously reported granular antigens of P-ANCA.
Biochemical cure of recurrent acromegaly by resection of cervical spinal canal metastases
Authors: Dayan Colin , Guilding Tess , Hearing Stephen , Thomas Paul , Nelson Richard , Moss Tim , Bradshaw John , Levy Andy , Lightman Stafford
Published: November 2003

Pituitary carcinoma with metastatic endocrine secreting tissue is rare. Eight cases of malignant, growth hormone‐secreting tumours, all metastatic within the central nervous system have been previously described. Complete surgical resection was not possible and the patients died within 4 years of presentation with distant spread. Here we describe the first case of an apparent surgical cure of a somatotroph carcinoma metastatic to the cervical spine, documented by biochemical assessment and magnetic resonance and radioligand imaging. The possibility that intrathecal metastasis of somatotroph tumours may be responsible for some cases of treatment resistant acromegaly is discussed.
Coexistence of bilateral adrenal phaeochromocytoma and Idiopathic hyperaldosteronism
Authors: Tan Gerry H. , Aidan Carney J. , Grant Clive S. , Young Jr William F.
Published: November 2003

The coexistence of phaeochromocytoma and primary aldosteronism is rare. In the four cases reported in the English literature, adrenal gland involvement was unilateral. The case we report is unusual in that both adrenal glands were affected. Our patient was a 63‐year‐old man who presented with long‐standing hypertension and hypokalaemia. Evaluation for secondary hypertension showed biochemical evidence of both phaeochromocytoma and primary aldosteronism. Bilateral adrenal masses were found on imaging studies. Right total and left partial adrenalectomy were performed. Pathological examination revealed bilateral phaeochromocytomas, with cortical findings consistent with mild cortical hyperplasia. Annual biochemical evaluation over a 2‐year period has shown no evidence of recurrent disease. To our knowledge, this is the first reported case in the English‐language literature of phaeochromocytomas involving both adrenal glands coexisting with hyperaldosteronism.
Functional integrity of granulosa cells from polycystic ovaries
Authors: Almahbobi Ghanim , Anderiesz Cleola , Hutchinson Paul , McFarlane James R. , Wood Carl , Trounson Alan O.
Published: November 2003

OBJECTIVE The polycystic ovarian syndrome is frequently associated with human infertility and is a partially characterized syndrome of unknown aetiology. The aim of this study was to describe the functional integrity of granulosa cells from polycystic ovaries.

PATIENTS Follicular aspirates were collected from polycystic ovaries of ovulatory (n = 24) and anovulatory (n = 7) patients. Follicular aspirates were also collected from normal ovaries of untreated (n = 24) and superovulated (n = 10) subjects. All patients were enrolled for the recovery of their oocytes for in vitro maturation and fertilization.

MEASUREMENTS FSH receptors and apoptosis were measured in the granulosa cells of the different patients. FSH‐stimulated oestradiol and LH‐stimulated progesterone production by granulosa cells of the different patients were also measured.

RESULTS The binding of 125I‐labelled human recombinant FSH to granulosa cells from anovulatory subjects with polycystic ovaries was significantly higher than that found in granulosa cells from normal (180%) and superovulated (163%) ovaries. However, the ligand binding to granulosa cells from ovulatory subjects with polycystic ovaries was not significantly higher than that found in normal granulosa cells. Also, granulosa cells obtained from anovulatory subjects with polycystic ovaries cultured with FSH produced more oestradiol than normal granulosa cells but oestradiol production was similar to that of granulosa cells from superovulated ovaries (mean ± SEM, 244.94 ± 22.02, 24.23 ± 2.92, 211.87 ± 50.39 nmol/l/24 h, respectively). Flow cytometric analysis showed that the proportions of viable and apoptotic granulosa cells (mean ± SDM, 70 ± 5 and 7 ± 1%, respectively) were similar in normal subjects and in those with polycystic ovaries.

CONCLUSION We conclude that most of the granulosa cells of polycystic ovaries are healthy and non‐apoptotic, expressing high levels of FSH receptors and highly responsive to this hormone in culture. These data provide direct evidence that most of the follicles of polycystic ovaries are not atretic.
Correlation of endoscopic cytology and histology in oesophageal cancer: results in Porto Alegre, RS—Brazil
Authors: FRANTZ M. A. , PROLLA J. C.
Published: November 2003

A retrospective study of oesophageal cytopathology at the Hospital de Clínicas de Porto Alegre (HCPA), RS, Brazil, from 1989 to 1992 was made to assess the sensitivity, specificity, predictive values and accuracy of endoscopic cytology and biopsy; and study the correlation between cytopathological and histopathological diagnosis. Specimens from 94 patients were available for review. The final diagnosis was based on surgical pathology and follow up. The 81 patients with cancer of the oesophagus had the following sex distribution: 64 males and 17 females (a 3.7–1 ratio). No tumour was found in 13 patients. The following conclusions were made: (i) there is excellent correlation between cytology and histology in oesophageal lesions sampled by endoscopy; (ii) a correct positive cytologic report was obtained in 77 (95%) of the 81 proven oesophageal cancers; a false‐negative or unsatisfactory result was given in four patients. A false‐positive diagnosis of cancer was not made. There were 13 true‐negative reports. These findings result in a sensitivity of 95% with 95% confidence intervals (CI) of 90.26–99.74%; a specificity of 100% (CI of 98.5–100%); a positive predictive value of 100% (CI of 99.3–100%); a negative predictive value of 76% (CI of 55.7–96.3%); (iii) a correct positive histological report was obtained in 67 (83%) of the 81 proven oesophageal cancers; a false‐negative or unsatisfactory result was given in 14 patients. A false‐positive diagnosis of cancer was not made. There were 13 true‐negative reports. These findings result in a sensitivity of 83% with 95% CI of 74.82–91.18%; a specificity of 100% (CI of 98.5–100%); a positive predictive value of 100% (CI of 99.25–100%); a negative predictive value of 48% (CI of 29.16–64.84%); (iv) of 81 patients with proven cancer, in 79 (98%) at least one of the methods was positive. In only two patients with cancer were both methods negative. These findings result in a combined sensitivity of 98% (CI of 94.92–100%); a specificity of 100% (CI of 98.5–100%); a positive predictive value of 100% (CI of 99.31–100%); and a negative predictive value of 87% (CI of 70–100%). Our series confirms the value of the combined use of cytology and biopsy for the investigation of oesophageal lesions. However, it should be remembered that even with the combined use of cytology and biopsy there are some tumours that will be negative by both procedures: we had only two such cases, confirming the rarity of such an event.
Phyllodes tumour: cytologic and histologic presentation of 22 cases, and immunohistochemical demonstration of p53
Authors: SHABALOVA I. P. , CHEMERIS G. J. , ERMILOVA V. D. , RODIONOVA L. M. , PAVLIKOVA N. A. , SYRJÄNEN K. J.
Published: November 2003

A retrospective study of 22 cases of phyllodes tumour (PT) was undertaken to evaluate the potential value of fine needle aspiration (FNA) cytology in the diagnosis of benign and borderline PT. Histological material was available from 12 patients with typical benign PT (group 1), six patients with less typical changes (group 2) and four cases of borderline PT (group 3). Cytological presentation of PT in these cases was similar to that described by other cytologists, although abundant cellular material was obtained in only eight FNAs, naked nuclei were present in nine cases only, and atypical or suspicious cytological features were found in seven cases. Comparative analysis of p53 was made in nine patients with PT, five cases with other benign breast lesions and five with malignant lesions. p53 reaction was positive in five of nine patients with PT (all cases from groups 2 and 3), compared with two of five cases of carcinoma. p53 was negative in all patients with PT from group 1 and the five other benign cases. We suggest that cytopathologists should be careful when a myxoid stromal component is present in cytological smears.
The cytokine‐mediated imbalance between coagulant and anticoagulant mechanisms in sepsis and endotoxaemia
Authors: LEVI M. , VAN DER POLL T. , TEN CATE H. , VAN DEVENTER S. J. H.
Published: November 2003

Disseminated intravascular coagulation (DIC) is a frequently occurring complication of sepsis and may contribute to multiple organ failure. More insight into the pathogenesis of this derangement of the coagulation system is necessary to develop more effective therapeutic strategies for this condition. Recently, more detailed knowledge on the pathogenetic pathways involved in DIC has been obtained by the study of models of experimental bacteraemia and endotoxaemia in human subjects and non‐human primates. The mechanisms that lead to activation of coagulation, potentiated by the simultaneous depression of physiological inhibitory systems and to impaired function of the fibrinolytic system, are outlined in this review. In addition, the mediatory role of various cytokines in the derangement of coagulation is discussed.
Gamma‐interferon inhibits Pneumocystis carinii attachment to lung cells by decreasing expression of lung cell‐surface integrins
Authors: POTTRATZ S. T. , WEIR A. L.
Published: November 2003

Pneumocystis carinii (PC) is a leading cause of pneumonia in immunocompromised patients. Previous work has shown that fibronectin (Fn) and Fn‐binding integrins mediate PC attachment to lung cells in vitro. Gamma‐interferon (γ‐IFN) is a major factor in host defence against PC infection. To determine the effect of γ‐IFN on PC attachment to lung cells, the alveolar epithelial cell line A549 was incubated with γ‐IFN (0–104 U mL−1) and attachment of 51Cr‐labelled PC to the A549 cells was quantified. PC attachment was significantly decreased (P < 0.01) by addition of γ‐IFN with no evidence of injury to either the PC or A549 cells. Effects of γ‐IFN on PC attachment were observed after 24 h and reached a maximum after 48 h of incubation. To investigate the mechanism of this decrease, we examined integrin expression on γ‐IFN‐treated A549 cells. A549 cell expression of the α5 and β1 integrin subunits was decreased, whereas expression of the αv subunit was unchanged. Northern blot analysis showed a similar decrease in mRNA for the α5 and β1 integrins. Therefore, γ‐IFN‐mediated inhibition of PC infection may, in part, result from inhibition of PC attachment to alveolar epithelial cells caused by γ‐IFN‐induced decreases in alveolar integrin expression.
Apert's syndrome: differential in vitro production of matrix macromolecules and its regulation by interleukins
Authors: Bodo M. , Carinci F. , Baroni T. , Giammarioli M. , Bellucci C. , Bosi G. , Pezzetti F. , Becchetti E. , Evangelisti R. , Carinci P.
Published: November 2003

During embryonic development, variations in the composition of the extracellular matrix (ECM) macromolecules influence bone tissue differentiation. We present novel findings on the in vitro phenotypic expression of periosteal fibroblasts obtained from patients affected by Apert’s syndrome, a rare craniofacial malformation, and the effects that interleukins (ILs) induce on the phenotype. Apert fibroblasts synthesized greater quantities of glycosaminoglycans (GAGs) and intracellular type I collagen, and also produced more type III collagen and fibronectin. The amount of hyaluronic acid (HA) secreted by Apert fibroblasts was much higher than that secreted by normal fibroblasts, but, as the absolute values of heparan sulphate (HS), chondroitin sulphate (CS) and dermatan sulphate (DS) also rose in Apert media, the HA–sulphated GAG ratio was similar in the media obtained from both populations. Both ILs triggered elevations of HA in normal cells, although relative percentage secretion remained unaltered, but significantly reduced HA secretion by Apert cells. IL‐1 significantly increased CS in normal and Apert media, whereas IL‐6 enhanced HS and DS in media of both populations. HA–sulphated GAG ratio decreased in Apert media after IL treatment. Both ILs boosted fibronectin production by Apert fibroblasts, whereas IL‐1 increased type III but not type I collagen. Taken together, these data demonstrate that the synthesis and secretion of ECM macromolecules are markedly altered in Apert fibroblasts. The fact that treatment with ILs further modifies the Apert phenotype suggests that ILs may be implicated in the pathophysiology of the malformations during skull morphogenesis.
Physical laws of cholesterol gallstone fragmentation
Authors: NEUBRAND M. , GREINWALD I. , LOBENTANZER H. , PAUMGARTNER G. , HERMEKING H. , SAUERBRUCH T.
Published: November 2003

Efficient fragmentation is the most important prerequisite for successful treatment of gallstones by extracorporeally induced shock waves. No data are available on the amount of energy necessary for stone disintegration and on the threshold energy below which no further fragmentation occurs. We therefore performed an in vitro investigation on human cholesterol gallstones to elucidate physical laws governing shock‐wave lithotripsy. First, the focal pressure of the lithotripter was measured to calculate the energy traversing a stone. Second, 96 gallstones from 16 gall bladders were analysed with respect to physicochemical composition, radiological features and ultrasound before fragmentation was performed. Energy for stone disintegration was constant within each stone family but varied between 4.6 J mL−1 and 36.8 J mL−1 in different families. This energy correlated linearly with stone volume. None of the radiological and physicochemical factors revealed a clear‐cut correlation of the different energies necessary for similar stone disintegration. The threshold energy differed between 0.26 mJ and 1.04 mJ per pulse. In conclusion, stone volume was the best parameter predicting stone fragmentation. However, in cholesterol stones with a similar composition the required energy per volume varies considerably together with the threshold energy. Radiological and ultrasound parameters appear to be of minor importance in explaining these differences.
Position‐dependent reproductive success of flowers in Dactylorhiza maculata (Orchidaceae)
Authors: Vallius E.
Published: November 2003

1. This study explores the importance of flower position for reproductive success of individual flowers of the deceptive terrestrial orchid, Dactylorhiza maculata (L.) Soó.

2. Upper flowers had smaller lips and produced lighter pollinia than those in other parts of the inflorescence, probably due to architectural effects.

3. Dry weight of seed capsules decreased from bottom to top in the inflorescence in both open‐pollinated and hand‐pollinated plants. However, removal of flowers from the middle and lowest parts of the inflorescence considerably increased seed production of the upper flowers. Decreased seed set was due to decreased resource availability because of the long maintenance time of opened but not pollinated lower flowers.

4. Experimental self‐pollination resulted in reduced seed set compared to that following pollination with cross‐pollen, but decreased seed set was independent of flower position within an inflorescence.

5.  Dactylorhiza maculata with large inflorescences did not have better pollination success. The production of more flowers than the plant is able to fully support may have evolved as an ovary reserve in case of flower herbivory, or as a ‘bet‐hedging’ strategy through which the plant can increase seed production in years with good pollinator availability.
Do endogenous seasonal cycles of food intake influence foraging behaviour and intake by grazing sheep?
Authors: Iason G. R. , Sim D. A. , Gordon I. J.
Published: November 2003

1. Large herbivores living in temperate regions show different degrees of seasonal biological variation, including voluntary food intake (VFI). The decline of VFI in winter has been hypothesized to be an evolved response to lower food availability or quality, which can act as an internal constraint on food intake.

2. The hypotheses were tested that (i) animals that have a greater inherent seasonal variation of VFI, measured indoors under ad libitum conditions, would also have a greater seasonal variation in intake and grazing behaviour under field conditions, and (ii) greater seasonal variation in intake and grazing behaviour under field conditions would be expressed at a higher level of food availability.

3. The intake and grazing behaviour in summer and winter, of three breeds of sheep, were compared at two levels of food availability (at pasture heights of 3·7 and 5·4 cm). The breeds were known to have contrasting degrees of seasonal variation in food intake when fed ad libitum; the VFI of the Shetland (SH) and Scottish Blackface (BF) sheep varies greatly between seasons whereas that of the Dorset Horn (DH) is less seasonally variable.

4. All three breeds consistently increased their rates of biting and duration of grazing activity in the winter, taking many more smaller bites each day than in the summer, and both digestibility and intake were lower in winter than in summer.

5. Contrary to expectation, the DH ewes had the highest seasonal difference of dry matter intake at pasture, whereas the SH breed had the lowest variation of intake between seasons.

6. This experiment provides no evidence that differences between seasons in intake and foraging behaviour in the field vary with the animals’ degree of endogenous seasonal variation in VFI. Variation between seasons was consistent at both levels of resource availability, suggesting that it resulted from seasonal changes in food quality (digestibility) rather than biomass availability. It is not easy to extrapolate from laboratory feeding studies, where animals’ own physiological constraints apply, to foraging ecology in the field, where constraints imposed by the environment may be more important.
HsMCM6: a new member of the human MCM/P1 family encodes a protein homologous to fission yeast Mis5
Authors: Tsuruga Hiromichi , Yabuta Norikazu , Hosoya Shiki , Tamura Kazutoshi , Endo Yuichi , Nojima Hiroshi
Published: November 2003

Background:

The tight regulatory mechanism that prevents more than one round of chromosomal DNA replication per cell cycle is thought to require the function of Mcm/P1 proteins. We report here the structural and functional analyses of HsMcm6, a human homologue of the Mis5 of Schizosaccharomyces pombe.

Results:

We demonstrate here that the transcription of the HsMCM6 gene was repressed in quiescent cells but was rapidly induced at the G1/S phase by growth factor stimulation. The 5′ regulatory region of the HsMCM6 gene was found to harbour four putative E2F binding motifs, and these were responsible for the promoter activity. The HsMcm6 protein level oscillated during the cell cycle, with a peak at the G1/S phase. We also showed that the cell‐cycle dependent change of subcellular localization of HsMcm6 resembles those of other Mcm/P1 proteins. HsMcm6 consists of two forms, a form extractable by Nonidet P‐40 and the nucleus‐bound form. A demonstration of the association of HsMcm6 with HsMcm2 and HsMcm7 in vivo supports the idea that they behave as a heteromeric complex. We mapped the HsMCM6 gene at 2q12–14.

Conclusion:

The results indicate that the behaviour of HsMcm6 is reminiscent of replication licensing factor like other Mcm/P1 family members.
DAP‐1, a novel protein that interacts with the guanylate kinase‐like domains of hDLG and PSD‐95
Authors: Satoh Kiyotoshi , Yanai Hiroyuki , Senda Takao , Kohu Kazuyoshi , Nakamura Tsutomu , Okumura Nobuaki , Matsumine Akihiko , Kobayashi Shigeru , Toyoshima Kumao , Akiyama Tetsu
Published: November 2003

Background:

The human homologue of the Drosophila discs large tumour suppressor protein (hDLG) and closely related proteins such as postsynaptic density protein 95 kDa (PSD‐95) are associated with N‐methyl‐d‐aspartate receptors (NMDA‐R) and Shaker‐type K+ channels, and are thought to be involved in their clustering.

Results:

We have identified a protein named DAP‐1 that binds to the guanylate kinase‐like domains of hDLG and PSD‐95. DAP‐1 was found to associate with hDLG, PSD‐95, NMDA‐R and adenomatous polyposis coli protein (APC). Furthermore, we found that DAP‐1 is specifically expressed in the brain and colocalizes with PSD‐95 and APC in mouse cerebellum. We also found that DAP‐1 is colocalized with PSD‐95 and NMDA‐R at the synapses in cultured rat hippocampal neurons.

Conclusion:

Our findings suggest that DAP‐1 may play several roles in the molecular organization of synapses and neuronal cell signalling by interacting with hDLG and PSD‐95, which in turn are associated with receptors, ion channels and APC.
Phage DNA packaging
Authors: Fujisawa Hisao , Morita Miyo
Published: November 2003

Phage DNA packaging occurs by DNA translocation into a preformed protein shell—a prohead—with the aid of a packaging enzyme or a terminase. The packaging enzyme is composed of two subunits: the large subunit has ATP‐binding, prohead binding, and DNA cleavage activities, and the small subunit is a DNA binding protein. DNA translocation is driven by ATP hydrolysis. In general, phage DNA replication mechanisms lead to the accumulation of concatemers. Concatemers are processed to mature DNA during and depending upon DNA packaging. This review will focus on the molecular mechanism of concatemer processing and the coupling of ATP hydrolysis to DNA translocation.
Saccharomyces cerevisiae recA homologues RAD51 and DMC1 have both distinct and overlapping roles in meiotic recombination
Authors: Shinohara Akira , Gasior Stephen , Ogawa Tomoko , Kleckner Nancy , Bishop Douglas K.
Published: November 2003

Background:

Rad51 and Dmc1 are Saccharomyces cerevisiae homologues of the Escherichia coli recombination protein RecA. Mutant analysis has shown that both proteins are required for normal meiotic recombination, for timely and efficient formation of synaptonemal complex and for normal progression out from meiotic prophase.

Results:

We have further characterized rad51 and dmc1 single mutants. A dmc1 mutation confers more severe defects in double strand break (DSB) resolution, crossover recombination and meiotic progression than does a rad51 mutant; in contrast, during return to growth, a rad51 mutation confers more severe defects in viability and intrachromosomal recombination than does a dmc1 mutation. Analysis of a rad51 dmc1 double mutant, in parallel with single mutants, shows that the double mutant is more defective with respect to the formation of crossovers during meiosis and, especially strikingly, with respect to interhomologue and intrachromosomal recombination during return to growth. Consistent with the observation of DMC1‐dependent recombination in a rad51 mutant, subnuclear complexes of Dmc1 protein were detected for the first time in this mutant. In contrast to the effects on recombination, the effect of the double mutant on meiotic progression was similar to that of the rad51 single mutant.

Conclusion:

Rad51 and Dmc1 each make unique contributions to meiotic recombination. However, the two proteins are capable of substituting for one another under some circumstances, implying that they most likely share at least one recombination function. Recombination and cell cycle phenotypes are all consistent with the possibility that a dmc1 mutation causes an arrest of the post‐DSB recombination complexes at a later, more stable stage than does a rad51 mutation.
Posterior colpoceliotomy: a technical variation in minimally invasive myoma surgery
Authors: Tuttlies F. , Keckstein G. , Ulrich U.
Published: November 2003

This report describes our experience in colpoceliotomy for myoma extraction following laparoscopic enucleation. Between September 1991 and August 1992 about 17 colpoceliotomies were performed. Patient follow‐up varied from 2 to 11 months. No serious complications were seen. Large myomas up to 11 cm in diameter and, in combination with the endobag, even multiple myomas or other specimens can be removed using this technique.
A new direction in laparoscopic suturing and knot tying
Authors: Murphy D. L.
Published: November 2003

An intracorporeal knot can be tied using instruments, employing a method that mimics the actions of the hooked index finger as used in the one‐handed, hand tie, of open surgery. The short end of the suture, when held in the instrument jaws, can be looped around the bite section of the suture. This rotation is facilitated by a spiral spur member, which is affixed to the instrument jaw, and which captures the bite. For maximum efficiency one uses paired left and right instruments, where the spiral spurs are directed clockwise and anticlockwise, respectively. Instrument rotation and minimal instrument transfers are required for this technique. The surgical half‐hitch can be created by two entirely different actions, the instrument‐ or the hand‐tied method. Each of these methods could be considered to be the converse of the other. Ergonomically designed instruments are essential for laparoscopic surgery. A role for instrument axial rotation is recognized and extended in this new method of laparoscopic knot tying. This novel technique contrasts with all known knotting methods.
Return to work following laparoscopic hysterectomy
Authors: Rosen D. M. B. , Cario G. M. , Carlton M. A. , Lam A. M.
Published: November 2003

Objective

To determine an accurate figure for the time of expected return to work or normal activity following a laparoscopic hysterectomy.

Design

A telephone survey of 100 patients who had undergone laparoscopic hysterectomy between September 1994 and July 1996.

Results

The mean time for return to work or normal activity was 3.3 weeks. Amongst those who worked, this figure was reduced to 2.9 weeks. Patients also reported that they could have returned to work at 2.3 weeks on average. These differences were statistically significant. There was also a significant difference between the recovery times of those reporting some complication and those who did not.

Conclusion

This study allows the surgeon to confidently advise patients of expected recovery times following laparoscopic hysterectomy. It also highlights the benefits of preoperative counselling, both in terms of patient motivation to recover and in terms of forewarning of minor complications and the effect of such complications on recovery time and therefore patient satisfaction.
Skin chemical suture using Histoacryl in laparoscopic surgery
Authors: Ghirardini G. , Battioni M. , Radighieri G.
Published: November 2003

Objective

To verify whether the use of Histoacryl tissue adhesive could replace skin suture of abdominal wounds in laparoscopy.

Design

Histoacryl was used to repair abdominal wounds in 60 consecutive cases of diagnostic or operative laparoscopy.

Results

The procedure was successful in 59 of the 60 cases.

Conclusions

Our data suggest that this method reduces the time needed for suture, and shows better aesthetic results and patient acceptance.
A case of malignant struma ovarii
Authors: Yamada T. , Saeki M. , Okuda K.
Published: November 2003

We report the case of a 31‐year‐old nulligravida who underwent laparoscopic ovarian cystectomy on account of the tentative diagnosis of mature ovarian cystic teratoma. However, malignant struma ovarii was suggested postoperatively. Additional conservative surgery was performed after 2 weeks, but no residual focus was detected. There has been no sign of recurrence during a 1‐year follow‐up period.
Laparoscopic‐assisted hysterectomy: disposable staples or diathermy?
Authors: Molloy D. , Doody Mary Lou
Published: November 2003

A total of 114 patients underwent laparoscopic‐assisted vaginal hysterectomy in a 38‐month period from December 1991, to February 1994. In 65 patients, disposable staples were used to achieve pedicle haemostasis and in 49 cases bipolar diathermy was used. No learning curve cases were excluded. Three additional patients were converted to open hysterectomy. No difference in outcome for the two groups was noted. Average operating time was slightly longer when diathermy was used. Average postoperative bed days were 2.8 days for the staple group and 2.7 days for the diathermy group. The cost‐effectiveness of the procedure may be improved when non‐disposable equipment is used for pedicle haemostasis.
Experience with combined use of laparoscopy and hysteroscopy for the investigation of infertility
Authors: Saridogan E. , Bolaji I. I. , Boto T. , Djahanbakhch Ovrang
Published: November 2003

Objective To present our experience with the combined use of laparoscopy and hysteroscopy as the primary investigation for tubal and uterine factors in infertility. Design This is a prospective study of patients investigated for infertility using hysterolaparoscopy for evaluation of infertility. Setting Assisted Conception and Infertility Unit at Newham General Hospital, London. Subjects These were 117 consecutive patients, of whom 53 (45.3%) had primary and 64 (54.7%) had secondary infertility. Results Laparoscopic examination revealed at least one abnormal finding in 77 (66.3%) patients and 29 (26.6%) patients had abnormal findings at hysteroscopy. Concurrent laparoscopy and hysteroscopy were completed in 108 patients in whom eight (7.4%) had abnormal hysteroscopic findings but normal laparoscopic findings. Five out of six (83.3%) patients who had intrauterine adhesions had a history of previous uterine surgery. Out of 67 patients with abnormal laparoscopy, 21 (31.3%) had abnormal findings at hysteroscopy compared with eight out of 41 (20.0%) with normal laparoscopy (χ2 = 1.678; P > 0.05). There was no significant difference in the prevalence of intrauterine abnormality detected hysteroscopically between the patients with primary and secondary infertility (χ2 = 2.899; P > 0.05). Conclusions The findings of this study suggest that routine use of hysteroscopy adds little information to the management of infertile patients and its concomitant use should be based on the history or physical examination findings. However, certain intrauterine abnormalities do not have warning signs. Until the significance of these intrauterine abnormalities in infertility is proven, routine use of hysterolaparoscopy for infertility investigation will remain a controversial issue.
Laparoscopic‐assisted vaginal hysterectomy: a low‐complication hysterectomy
Authors: Trehan A.K.
Published: November 2003

Object To determine the safety of laparoscopic‐assisted vaginal (LAVH) in a district general hospital. Design Retrospective analysis. Setting District general hospital in the UK. Subjects A total of 119 LAVH patients, who were unsuitable for vaginal hysterectomy according to conventional British criteria. Result The mean operating time taken was 91 min. Mean duration of stay was 2.2 days. The overall complication rate was 7.5%; most complications were of a minor nature. Conclusions We feel that in experienced hands LAVH is a safe procedure and, within our indications, is the hysterectomy method of choice in the district hospital setting.
Epithelial hyperplasia of usual type expresses both S100‐α and S100‐β in a heterogeneous pattern but ductal carcinoma in situ can express only S100‐α in a monotonous pattern
Authors: ICHIHARA S. , KOSHIKAWA T. , NAKAMURA S. , YATABE Y. , KATO K.
Published: November 2003

Immunohistochemical identification of myoepithelial cells using α‐smooth muscle actin provides little information about the nature of solid or quasi‐solid portions of epithelial hyperplasia and ductal carcinoma in situ (DCIS) because actin‐rich myoepithelial cells are usually demonstrated only in the stromal–epithelial junction of both lesions. We studied the differential distribution of α‐subunit (S100‐α) and β‐subunit (S100‐β) of S100 protein in actin‐negative areas of usual epithelial hyperplasia and DCIS by employing the streptavidin method with monospecific rabbit antibodies against each subunit. All usual epithelial hyperplasias (n=17) were composed of heterogeneous epithelial cell types; cells expressing S100‐α and/or S100‐β were intermingled with non‐expressing cells, resulting in a mosaic‐like pattern. On the contrary, DCIS (n=32) uniformly lacked immunoreactive S100‐β; S100‐α was diffusely expressed in 24 (68.8%) DCIS (three solid/comedo, 13 cribriform, four endocrine, one micropapillary, three papillary variants) and negative in the remaining eight (31.2%) DCIS (one cribriform, two micropapillary, four papillary and one apocrine variants). In conclusion, in contrast to usual epithelial hyperplasia that expresses both S100‐α and S100‐β in a heterogeneous pattern, DCIS can express only S100‐α in a monotonous pattern, possibly signifying unidirectional differentiation toward secretory glandular epithelium.
Prognostic comparison of three classifications for medullary carcinomas of the breast
Authors: LIDANG JENSEN M. , KIÆR H. , ANDERSEN J. , JENSEN V. , MELSEN F.
Published: November 2003

The aim of this study was to make prognostic comparisons between the modified scheme of Pedersen et al. the definitions of Tavassoli and the Ridolfi criteria for medullary carcinomas. Sixty breast carcinomas primarly diagnosed as medullary carcinomas were reclassified into typical medullary carcinoma (TMC), atypical medullary carcinoma (AMC) and non‐medullary carcinoma (NMC) according to the three classifications. The Ridolfi classification proved to be superior to the two other schemes in discriminating survival differences between the three groups TMC, AMC and NMC. All 13 patients with TMC are still alive indicating an excellent prognosis, while 29% and 39% of the 47 patients in the AMC and NMC category, respectively, have died of their disease. In the simplified system of Pedersen et al. the survival at 10 years for TMC patients decreased to 75% and no significant survival difference between the three groups could be demonstrated. As the prognosis for AMC proved to be worse compared to TMC and in fact was similar to NMC with values of 43% at 10 years in the Ridolfi classification, we find no reasons to maintain this category. We conclude that as long as no alternative and more easily applicable diagnostic method exists, pathologists should still apply the Ridolfi criteria on these tumours with medullary features leaving two diagnostic possibilities: TMC or NMC (i.e. poorly differentiated ductal carcinoma). Only lesions that fulfil all six criteria without any doubt should be diagnosed as TMC, thus avoiding overdiagnosis and a resulting risk of undertreatment.
Aggressive angiomyxoma: reappraisal of its relationship to angiomyofibroblastoma in a series of 16 cases
Authors: GRANTER S.R. , NUCCI M.R. , FLETCHER C.D.M.
Published: November 2003

Aggressive angiomyxoma is a distinctive soft tissue tumour associated with a high risk of local recurrence but lacks metastatic potential. This tumour occurs nearly exclusively in the soft tissues of the pelvis and perineum of adult women. The line of differentiation is not firmly established, but a fibroblastic/myofibroblastic origin has been proposed. We report 16 new cases of aggressive angiomyxoma of the pelvic soft tissue in women. In all cases bundles of cells, most often adjacent to vessels, with histological features of smooth muscle cells were identified. In 11 of 14 cases the myoid bundles were immunoreactive for desmin; they were also postive for smooth muscle actin in 10 of 11 cases. In 13 of 14 cases lesional stromal cells showed immunoreactivity for desmin. Three cases showed areas with histological features similar to those of angiomyofibroblastoma of the vulva, thus representing previously undescribed morphological overlap between these two entities. We conclude that aggressive angiomyxoma and angiomyofibroblastoma are related neoplasms in a spectrum of tumours showing myofibroblastic origin. Furthermore, the demonstration of immunoreactivity for desmin in aggressive angiomyxomas implies that this antibody is not helpful in discriminating between these two tumours, and the principal means of distinction remains histomorphological analysis.
Expression of major histocompatibility complex class II antigens on normal porcine intestinal endothelium
Authors: WILSON A. D. , HAVERSON K. , SOUTHGATE K. , BLAND P. W. , STOKES C. R. , BAILEY M.
Published: November 2003

A novel monoclonal antibody (MIL 11) specific for an antigen expressed on porcine endothelial cells is described. The antigen recognized by MIL 11 is most strongly expressed in the intestine but is also expressed on the capillary endothelium of a wide range of tissues. Using two‐ and three‐colour immunofluorescence microscopy we demonstrated the extensive coexpression of MIL 11 and major histocompatibility complex (MHC) class II antigens on normal porcine capillary endothelium in the intestine, trachea, thymus and small veins, while endothelium of large vessels and the heart were negative for MHC class II. In contrast to humans and rodents, available reagents do not detect MHC class II on the intestinal epithlium of pigs. However, porcine intestinal endothelium expressed both DR and DQ antigens. A population of strongly class II‐positive cells was also detected immediately adjacent to the endothelium in the lamina propria. Three‐colour immunofluorescence microscopy highlighted the close association between endothelium and intestinal CD4+ T cells. Lamina propria T cells were mainly MHC class II positive, whereas those in the epithelial compartment were MHC class II negative.
Mechanisms of pertussis toxin‐induced myelomonocytic cell adhesion: role of Mac‐1(CD11b/CD18) and urokinase receptor (CD87)
Authors: WONG W. S. F. , SIMON D. I. , ROSOFF P. M. , RAO N. K. , CHAPMAN H. A.
Published: November 2003

Stimulation of monoblastic U937 cells with transforming growth factor β1 and 1,25‐(OH)2 vitamin D3 (TGF‐β1/D3) upregulates urokinase receptor (uPAR) and confers urokinase‐dependent adhesiveness to the cells for serum‐or vitronectin‐coated surfaces. Recent studies show that uPAR itself is a high‐affinity adhesion receptor for vitronectin and that urokinase (uPA) is an activator of this adhesive function. In the course of exploring possible G‐protein involvement in this adhesion it was observed that TGF‐β1/D3‐primed U937 cells became adhesive to vitronectin in an uPAR‐dependent manner when exposed to pertussis toxin (PTX). The adherent response is concentration‐ and time‐dependent, and was not due to the ADP‐ribosyltransferase activity of the toxin because the purified B‐subunit of PTX was equally effective. Although promoting adhesion to serum‐ or vitronectin‐coated surfaces, PTX blocked spontaneous cell adhesion to fibrinogen, an endogenous ligand for the Mac‐1 receptor (CD11b/CD18). Flow cytometry study showed that expression of the α‐subunit of Mac‐1 (CD11b) on primed cells was increased by nearly threefold. Monoclonal antibody to CD11b abolished the PTX‐induced cell adhesion and the binding of the primed cells to PTX‐coated plates. Activation of Mac‐1 receptor by its endogenous ligand fibrinogen induced cell adherent response similar to PTX. PTX, but not uPA, triggered a rapid rise in [Ca2+]i in primed U937 cells, and PTX‐induced adhesion was significantly attenuated by 1,2‐bis‐(2‐aminophenoxy)ethane‐NNN ′,N ′‐tetraacetic acid/acetoxy‐methyl ester (BAPTA/AM), a selective membrane‐permeant [Ca2+]i chelator. PTX‐induced cell adhesion was also prevented by antibodies to uPAR and by conditioned medium containing soluble uPAR. Together these data indicate that PTX B‐subunit may bind to Mac‐1 integrin, which leads to a rapid rise in [Ca2+]i and subsequent activation of uPAR for adherence to vitronectin, suggesting a functional link between Mac‐1 and activation of uPAR important to cellular trafficking and host defence in response to Bordetella pertussis infection.
Molecular mechanism of T‐cell control of Chlamydia in mice: role of nitric oxide in vivo
Authors: IGIETSEME J. U.
Published: November 2003

T‐cell‐mediated immunity is crucial for the control of Chlamydia in mice. Recent evidence from studies in an in vitro model of the mucosal epithelium, the polarized epithelial–lymphocyte co‐culture (PELC) system, indicated that protective murine T cells mediated intracellular inhibition of the Chlamydia trachomatis agent of mouse pneumonitis (MoPn) at least partly by activating the interferon‐γ (IFN‐γ)‐inducible nitric oxide synthase (iNOS) pathway. To investigate whether nitric oxide played a role in controlling chlamydial infection in vivo, the protective capacity of a chlamydial‐specific T‐cell clone (clone 2.14‐0) was analysed in mice in the presence of a specific inhibitor of iNOS. The results revealed that the ability of this clone to clear Chlamydia in vivo is in part mediated by induction of nitric oxide (NO) production. The L‐arginine analogue and iNOS inhibitor, NG‐monomethyl‐L‐arginine monoacetate (MLA), increased the chlamydial burden in infected mice and inhibited the ability of clone 2.14‐0 to clear genital MoPn infection in vivo. The results are consistent with the working hypothesis that the IFN‐γ‐inducible iNOS pathway is involved in the control of Chlamydia by T lymphocytes in mice.
Sequence, specificity and crystallization of an oestrone‐3‐glucuronide antibody (3910)
Authors: HE M. , GANI M. , LIVNAH O. , STURA E. A. , BEALE D. , COLEY J. , WILSON I. A. , TAUSSIG M. J.
Published: November 2003

We describe the specificity profile and V region sequences of a high‐affinity monoclonal antibody (mAb), 3910, directed against oestrone‐3‐glucuronide (E3G). Inhibition studies show that the D‐ring is critical for steroid specificity, while the glucuronic acid attached to the A ring is required for high binding affinity, suggesting that both ‘ends’ of the E3G ligand are recognized. The VH domain is encoded by a gene from the VH7183 family, while VL appears to be encoded by the Vk5.1 gene (κII subgroup) with a deletion of six residues from complementarity‐determining region‐1 (CDR1). The VH CDR3 is 10 amino acid residues in length, of which D/N contributes five residues. Comparison of VH CDR of 3910 with those of mAb against progesterone (DB3) and digoxin (26–10, 40–50), for which crystal structures have been determined, suggests that aromatic side chains are important for E3G binding and that tyrosine residues H50, H97 and H100 may interact with the ligand. The Fab fragment of 3910 has been crystallized in its native and steroid (E3G and oestriol‐3‐glucuronide) complexed forms. An X‐ray diffraction data set to 3 Å resolution has been collected for the native Fab.