bluuebunny/crossref_metadata_2025 huggingface dataset:
Seasonal field metabolic rate and dietary intake in Arabian Babblers (Turdoides squamiceps) inhabiting extreme deserts
Authors: Anava A. , Kam M. , Shkolnik A. , Degen A. A.
Published: November 2003

1. Arabian Babblers (Turdoides squamiceps Cretzsch.; mean adult body mass = 72·5 g) inhabit extreme deserts of Israel. They consume invertebrates and fruits and, at least at our study site, do not drink. It was hypothesized that babblers (1) in general, use relatively less energy and water than other birds of its body mass; and (2) consume a more water‐rich diet (mainly fruits) in summer and more energy‐rich diet (mainly invertebrates) in winter. Doubly labelled water was used to determine seasonal field metabolic rate (FMR) and water influx rate (WIR) and to estimate dietary selection in free‐living Arabian Babblers.

2. Babblers in winter weighed significantly more than in summer, and males weighed more than females in both seasons. Tritiated water (TOH) space, as a proportion of body mass, was higher in males than in females in summer but no difference between sexes was found in winter. Males in summer had a higher TOH space, proportionally, than males in winter but there was no difference between seasons in females. Mass‐specific WIR did not differ between sexes in any season and averaged 0·475 ml g−1 d−1 in winter which was significantly higher than the 0·283 ml g−1 d−1 in summer.

3. The mean daily energy expenditure of the babblers did not differ either between seasons or between sexes within seasons and averaged 1·61 kJ g−1 d−1 in winter and 1·68 kJ g−1 d−1 in summer. It was calculated that each babbler consumed an average of 5·09 g dry matter invertebrates and 1·83 g dry matter fruits in summer (for a 68·2‐g bird; mean adult body mass in summer) and 3·49 g dry matter invertebrates and 6·61 g dry matter fruits in winter (for a 76·9‐g bird; mean adult body mass in winter).

4. When compared with other avian species, FMR and WIR of babblers were lower than bird species in general, but were similar to those of other desert birds. It was calculated that proportional dietary intake, on a dry matter basis, included 0·79 insects and 0·21 fruits in summer and 0·35 insects and 0·65 fruits in winter. Therefore, the babblers consumed a relatively energy‐rich diet in summer and water‐rich diet in winter which refuted our hypothesis. Most of the metabolizable energy was provided by invertebrates in both seasons; invertebrates provided more water in summer but fruits provided more in winter.
Dynamics of chromosomes and microtubules visualized by multiple‐wavelength fluorescence imaging in living mammalian cells: effects of mitotic inhibitors on cell cycle progression
Authors: Haraguchi Tokuko , Kaneda Toru , Hiraoka Yasushi
Published: November 2003

Background:

Mitotic events are accomplished by a coordinated interaction of molecular components within a cell. The continuous observation of specific molecular components in individual living cells provides a unique opportunity to examine the temporal and spatial coordination of these components. In order to be able to observe dynamic events in living mammalian cells, we have developed a computerized fluorescence microscope system.

Results:

Using this fluorescence microscope system, we observed the dynamic behaviour of chromosomes and microtubules in living HeLa cells throughout an entire cell cycle. Chromosomes and microtubules were stained with Hoechst 33342 and rhodamine‐conjugated tubulin, respectively. Microinjected rhodamine‐tubulin was incorporated into microtubules at all stages of the cell cycle. In addition, rhodamine‐tubulin stained the centrosomes during the G2 phase of the cell cycle and allowed us to observe centrosome movement. We also examined the effects of mitotic inhibitors on cell cycle progression. In a living cell treated with an inhibitor of type II DNA topoisomerase (ICRF‐193) chromosomes formed an aberrant metaphase plate and failed to segregate. However, following mitotic spindle disassembly, the chromosomes decondensed normally. While the cell began to divide into two portions, cell division was eventually aborted and the daughter cells, still connected by a cytoplasmic bridge, rejoined into a single cell. These results demonstrate how a polyploid nucleus is generated in the presence of ICRF‐193. On the other hand, microtubule polymerization inhibitors, such as colchicine, nocodazole and vinblastine, arrested the cell in the metaphase with condensed chromosomes; cytochalasin D did not affect the mitotic behaviour of either chromosomes or microtubules.

Conclusions:

Our observations of living cells demonstrate that the failure of chromosome segregation in and of itself does not prevent an exit from the metaphase, whereas the inhibition of microtubule assembly arrests cells at metaphase. Thus, a metaphase checkpoint is monitored by functional spindle microtubules but not by the completion of chromosome segregation.
Alteration of telomeric sequences and senescence caused by mutations in RAD50 of Saccharomyces cerevisiae
Authors: Mary Kironmai K. , Muniyappa K.
Published: November 2003

Background:

Vegetatively dividing cells of Saccharomyces cerevisiae carrying a mutation in RAD50 grow significantly more slowly in rich medium and are sensitive to DNA damage inflicted by X‐ray or chemical mutagens. RAD50 function is essential for the formation and repair of meiosis‐specific double‐strand breaks and chromosome stability.

Results:

We present evidence for two new phenotypes associated with the rad50Δ mutant; shortened telomeres and cell senescence. Comparison of TG1–3 telomeric sequences in an isogenic pair of RAD50 and rad50Δ haploid strains showed that they were considerably shortened in the latter. Although rad50Δ mutation conferred cell enlargement and slow growth, cell doubling was faster but caused an increase in the frequency of cell death. Telomeres were restored to the wild‐type size in hemizygous RAD50/rad50Δ and rad50S/rad50Δ strains; however, they showed a significant increase in rad50S/rad50S diploid with a concomitant rise in cell viability. Telomeres were stabilized in hemizygous RAD50/rad50Δ and rad50S/rad50Δ diploids during prolonged growth, suggesting that even a half‐dosage of RAD50 is sufficient to conserve the telomere size during successive cell divisions. Furthermore, cells bearing the rad50Δ mutation revealed abnormalities in nuclear segregation and, in the presence of hydroxyurea, displayed phenotypes consistent with defects in S‐phase checkpoint control.

Conclusion:

This report presents evidence of the involvement of a gene relevant to recombinational repair in the maintenance of telomeres.We conclude that the phenotypes displayed by yeast rad50Δ cells have intriguing similarities among the human cell lines representing DNA repair‐deficient chromosome instability syndromes.
Characterization of the mouse prostaglandin F receptor gene: a transgenic mouse study of a regulatory region that controls its expression in the stomach and kidney but not in the ovary
Authors: Hasumoto Ken‐yuh , Sugimoto Yukihiko , Gotoh Megumi , Segi Eri , Yamasaki Atsushi , Yamaguchi Masahiro , Honda Hiroaki , Hirai Hisamaru , Negishi Manabu , Kakizuka Akira , Ichikawa Atsushi
Published: November 2003

Background:

The actions of prostaglandin F2α are mediated by a cell‐surface receptor (FP), but little is known about the regulation of FP gene expression. To clarify the mechanisms underlying tissue specific transcription of the mouse FP gene, we isolated and characterized mouse genomic DNA clones encoding FP.

Results:

Structural analysis revealed that the mouse FP gene is composed of three exons and two introns, and spans more than 11 kilobases. By primer extension and PCR analyses, the major transcription start site was identified as a cytosine nucleotide, but additional sites of transcription initiation were found in the ovary. There was no apparent difference in the FP gene transcription initiation site between the ovary, kidney and stomach. Sequence analysis of the putative promoter region showed only two potential SP‐1 binding sites, but no other typical well‐known consensus sequences. We generated transgenic mice with the potential promoter region of the FP gene connected to the lacZ reporter gene. Northern blot analysis showed that the pattern of expression of the transgene corresponded to that of FP expression, except in the ovary. Upon analysis by in situ hybridization, the lacZ gene transcripts were found to be expressed in the fundic glands in the stomach, and the cortical tubules in the kidney, in which endogenous FP transcripts were also expressed. On the contrary, expression of lacZ transcripts was not detected in the corpora lutea, where the highest expression of FP mRNA was observed.

Conclusions:

These studies suggest that a separate control mechanism exists for FP expression in the ovary, distinct from the expression in the stomach and kidney.
Inhibition of transpositional recombination by OrfA and OrfB proteins encoded by insertion sequence IS3
Authors: Sekine Yasuhiko , Izumi Ken‐ichiro , Mizuno Takaaki , Ohtsubo Eiichi
Published: November 2003

Background:

An insertion element IS3 is flanked by terminal inverted repeat (IR) sequences. IS3 encodes two, out‐of‐phase, overlapping open reading frames, orfA and orfB, from which three proteins are produced. OrfAB is a transframe protein produced by −1 translational frameshifting between orfA and orfB, and it is known to be IS3 transposase. OrfA and OrfB are the proteins produced without frameshifting, but their functions have not been elucidated.

Results:

A plasmid carrying an IS3 mutant that produces only transposase generates miniplasmids—which are the IS3‐mediated intramolecular transposition products—as well as characteristic IS3 circles and linear IS3 molecules. OrfA inhibited the generation of these small molecules to a lesser degree, but OrfB did not. OrfB, together with OrfA, however, inhibited the generation more strongly than OrfA alone. OrfA also inhibited the intermolecular transposition of mini‐IS3 with the chloramphenicol‐resistance gene flanked by IRs to a reduced frequency, and OrfB together with OrfA inhibited it almost completely. OrfA and/or OrfB did not, however, repress transcription from the promoter in the left‐terminal region preceding orfA.

Conclusions:

The results obtained above show that OrfA and OrfB are not repressors but are inhibitors of transpositional recombination promoted by transposase. OrfA with an α helix‐turn‐α helix DNA‐binding motif may compete with transposase to bind to terminal IRs. OrfA, together with OrfB that has a DDE motif conserved in retroviral integrases, may inhibit the formation of an active transpososome consisting of transposase, two terminal IRs and target DNA for the strand transfer reaction. IS3 with a limited size, 1258 bp in length, uses strategies of translational frameshifting and coupling to produce transposase as well as negative regulators to make its copies at a low level, which minimizes a deleterious effect of transposition on bacterial hosts.
Chromatin assembly factors: a dual function in nucleosome formation and mobilization?
Authors: Ito Takashi , Tyler Jessica K. , Kadonaga James T.
Published: November 2003

Chromatin assembly is a process that interfaces DNA replication, gene expression and progression through the cell cycle, and it is therefore critically involved in many important biological phenomena. This brief review provides a general background to the study of chromatin assembly, as well as an overview of putative chromatin assembly factors. Interestingly, recent data suggest that the ATP‐utilizing chromatin assembly factor, ACF, functions not only in nucleosome formation, but also in the ATP‐dependent remodelling of chromatin that facilitates DNA‐utilizing processes, such as transcription, replication, recombination, and repair.
How representative are Pipelle endometrial biopsies? A retrospective analysis of 324 biopsies followed by transcervical resection of the endometrium or hysterectomy
Authors: Krampl E. , Søby B. , Istre O.
Published: November 2003

Objective

To evaluate the diagnostic accuracy of Pipelle endometrial biopsies before surgical treatment of uterine bleeding disorders.

Design

Retrospective analysis.

Setting

County Hospital unit specializing in operative hysteroscopy.

Interventions

Routine outpatient endometrium biopsies were taken from 628 patients because of uterine bleeding disorders. Of these patients, 249 underwent transcervical resection of the endometrium (TCRE) and 75 had a hysterectomy within 6 months.

Results

Comparing the two subgroups ‘malignant’ and ‘benign’, the positive predictive value for endometrial malignancy was 100% (95% CI, 73.5–100). Insufficient tissue was obtained in five cases (1.5%); in three there were submucous fibroids, in one sampling took place after TCRE, and one patient had atrophic endometrium.

Conclusion

Adequate preoperative histological assessment of the endometrium is feasible with Pipelle biopsies, and they are highly accurate in detecting endometrial malignancy.
Laparoscopic supracervical (subtotal) hysterectomy: a first series of 500 cases
Authors: Donnez J. , Nisolle M. , Smets M. , Polet R. , Bassil S.
Published: November 2003

From July 1990 to April 1996, 500 laparoscopic supracervical (subtotal) hysterectomies (LASH) were performed. The technique and the advantages are described here. Because of the feasibility of this technique, the quick recovery of the patient and low rate of complications, this strictly laparoscopic approach must be widely considered when a hysterectomy is indicated and if there is no risk of remaining cervical cancer.
Histological phenotypes of enteric smooth muscle disease causing functional intestinal obstruction in childhood
Authors: SMITH V.V. , MILLA P.J.
Published: November 2003

Aims: Functional intestinal obstruction or chronic idiopathic intestinal pseudo‐obstruction is due to defects either in the enteric innervation or in intestinal smooth muscle. We have studied full‐thickness intestinal biopsies from 27 patients with functional intestinal obstruction due to enteric smooth muscle disease by routine histology and electron microscopy together with histochemical and immunohistochemical techniques to detect changes in the intestinal smooth muscle. Methods and results: Two patients appeared to have an acquired intestinal myopathy as a result of an autoimmune process. In 25 the disorders were congenital, of these seven had segmental abnormalities limited to the rectum and distal colon and 18 had a diffuse disease affecting both the small and large bowel. We identified five apparent histological phenotypes of enteric muscle disease, three of which represent abnormalities in morphogenesis resulting in alterations in intestinal muscle layering and two exemplify intrinsic myocyte defects and/or changes in the extracellular matrix. Conclusions: Careful phenotyping of these patients is important in devising optimal treatment and in understanding the underlying defect as well as the possible genetic mechanisms resulting in these abnormalities. Recognition of autoimmune smooth muscle disease is helpful, since making the diagnosis influences the patient's management.
High‐grade papillary cystadenocarcinoma of the tongue
Authors: POLLETT A. , PEREZ‐ORDONEZ B. , JORDAN R.C.K. , DAVIDSON* M.J.
Published: November 2003

Aims: Salivary gland tumours of the tongue are rare. The most common type is low‐grade mucoepidermoid carcinoma followed by adenoid cystic carcinoma. Papillary cystadenocarcinoma of salivary glands are uncommon lesions with low‐grade histological and clinical features. We report a high‐grade papillary cystadenocarcinoma in an 80‐year‐old man who presented with a tongue mass and metastatic disease in the neck. Methods and results: He was treated with partial glossectomy and bilateral neck dissection but developed local and regional recurrences 6 months later. The tumour had a prominent cystic appearance and had areas of necrosis. The cyst lumen was occupied by numerous papillae lined by pseudostratified columnar cells with a high nuclear–cytoplasmic ratio. The cytoplasm was eosinophilic, the nuclei were pleomorphic and exhibited irregular nuclear membranes, vesicular chromatin and prominent eosinophilic nucleoli. The mitotic activity was high and there were occasional abnormal mitotic figures. Metastatic carcinoma was present in four lymph nodes. The differential diagnosis of this unusual lesion includes cystadenoma, salivary duct carcinoma and metastases. Conclusions: This case and a review of the literature indicates that papillary cystadenocarcinomas of salivary gland origin exhibit a wider morphologic spectrum than described in the latest World Health Organization (WHO) classification which defines these lesions as low‐grade neoplasms.
Multifocal alveolar hyperplasia associated with lymphangioleiomyomatosis in tuberous sclerosis
Authors: LANTUEJOUL S. , FERRETTI G. , NEGOESCU A. , PARENT B. , BRAMBILLA E.
Published: November 2003

Multifocal alveolar hyperplasia associated with pulmonary lymphangioleiomyomatosis is reported in a 21‐year‐old woman with tuberous sclerosis. Beside the cystic lesions of lymphangioleiomyomatosis, the tomography showed nodules up to 8 mm in both upper lobes. A proliferation of type II pneumonocytes and Clara cells lining the alveolar walls in an adenoma‐like pattern was observed. Nuclear atypia, mitoses and necrosis were not observed, providing evidence against multicentric bronchioloalveolar carcinoma or micronodular atypical alveolar adenomatous hyperplasia. Whereas the lymphangioleiomyomatosis lesions showed strong positivity for HMB45 and expressed oestrogen and progesterone receptors, the alveolar hyperplasia was negative for these markers as it was for carcinoembryonic antigen, p53 and MIB1 antibodies. Multifocal alveolar hyperplasia in tuberous sclerosis is probably a benign hamartomatous lesion in our case without progression on a 2‐year follow‐up. Its histogenesis is unknown, but is possibly related to chromosome instability.
Quantification of immunocompetent cells in testicular germ cell tumours
Authors: TORRES A. , CASANOVA J.F. , NISTAL M. , REGADERA J.
Published: November 2003

The immunocompetent cells present in the different histological patterns of 43 testicular germ cell tumours were evaluated. CD3 + and CD45RO + (UCHL1 +) T lymphocytes, CD68 + and MAC 387 + macrophages, CD20 + (L26 +) B lymphocytes, and kappa and lambda + plasma cells were counted. The number of immunocompetent cells per mm2 of tumour tissue, excluding the necrotic areas, was evaluated. Microscopic fields were randomly selected by two observers. In order to guarantee randomization each surface was divided into parts, numbered through a lattice, and some fields were chosen via a random numbers table. This procedure yielded significantly different counts from those obtained on subjective selection. The number of T‐lymphocytes and macrophages was higher in seminomas than in the non‐seminomatous testicular germ cell tumours (P < 0.05) Embryonal carcinomas had more T‐lymphocytes than immature teratomas. No significant differences were found among testicular germ cell tumours with regards to the B‐lymphocytes, with the exception of the high number of B‐lymphocytes in mature teratomas. Kappa + and lambda + plasma cells were few in the testicular germ cell tumours. Randomization in the quantification of immunocompetent cells in testicular germ cell tumours is a good means for evaluation of immune response in all the tumour mass, not only in the areas with the most intense inflammatory cell infiltrate, and permits comparison of testicular germ cell tumours with other malignant tumours. Study of immunocompetent cells in every histological type of testicular germ cell tumour is useful in comparing them with other extra‐testicular germ cell tumours.
Chronic lymphocytic sialoadenitis in HCV‐related chronic liver disease: comparison with Sjögren's syndrome
Authors: SCOTT C.A. , AVELLINI C. , DESINAN L. , PIRISI M. , FERRACCIOLI G.F. , BARDUS P. , FABRIS C. , CASATTA L. , BARTOLI E. , BELTRAMI C.A.
Published: November 2003

With the aim of morphologically characterizing chronic sialoadenitis in patients with hepatitis C virus (HCV) chronic liver disease, labial salivary gland biopsies from 22 chronic HCV liver disease and from 10 primary Sjögren's syndrome patients were compared. Only focus score (number of aggregates with more than 50 lymphocytes per 4 mm2 of glandular tissue) and grading of inflammation were able to discriminate significantly between the two patient groups. Duct ectasia, acinar depletion, presence of lymphoid aggregates with less than 50 lymphocytes and of lymphoid infiltration within intralobular salivary duct epithelium were evident in both disease groups and appeared to be non‐specific, mostly age‐related changes. In both patient groups plasma cell and lymphocyte typing showed similar features: T lymphocytes represented most of the lymphoid population, B lymphocytes were few unless follicles were present. Higher focus score values were associated with a plasma cell switch from an IgA to an IgM and/or IgG predominance. A greater morphological similarity was seen between biopsies of the primary Sjögren's syndrome group and those of female rather than male chronic HCV liver disease patients. Salivary gland tissue in HCV patients responds to damage in a fashion similar to primary Sjögren's syndrome, the only difference being a lesser degree of inflammation.
Novel polymer‐grafted starch microparticles for mucosal delivery of vaccines
Authors: HERITAGE P. L. , LOOMES L. M. , JIANXIONG J. , BROOK M. A. , UNDERDOWN B. J. , MCDERMOTT M. R.
Published: November 2003

Recent studies have demonstrated that systemic and mucosal administration of soluble antigens in biodegradable microparticles can potentiate antigen‐specific humoral and cellular immune responses. However, current microparticle formulations are not adequate for all vaccine antigens, necessitating the further development of microparticle carrier systems. In this study, we developed a novel microparticle fabrication technique in which human serum albumin (HSA) was entrapped in starch microparticles grafted with 3‐(triethoxysilyl)‐propyl‐terminated polydimethylsiloxane (TS‐PDMS), a biocompatible silicone polymer. The immunogenicity of HSA was preserved during the microparticle fabrication process. Following intraperitoneal immunization of mice, TS‐PDMS‐grafted microparticles (MP) dramatically enhanced serum IgG responses compared with ungrafted MP and soluble HSA alone (P < 0.001). When delivered orally, both TS‐PDMS‐grafted and ungrafted microparticles elicited HSA‐specific IgA responses in gut secretions, in contrast to orally administered soluble antigen. Indeed, TS‐PDMS‐grafted microparticles stimulated significantly stronger serum IgG (P < 0.005) and IgA (P < 0.001) responses compared with those elicited by ungrafted microparticles. These findings indicate that TS‐PDMS‐grafted starch microparticles have potential as systemic and mucosal vaccine delivery vehicles.
Modulation of WC1, a lineage‐specific cell surface molecule of γ/δ T cells, augments cellular proliferation
Authors: HANBY‐FLARIDA M. D. , TRASK O. J. , YANG T. J. , BALDWIN C. L.
Published: November 2003

WC1, also known as T19, is the only unique γ/δ T‐cell differentiation antigen described to date other than the γ/δ T‐cell receptor. We present evidence that modulation of WC1 results in augmented proliferation of γ/δ T cells. Immobilized IL‐A29, a monoclonal antibody (mAb) specific for WC1, augmented proliferation of γ/δ T cells in the autologous mixed leucocyte reaction (AMLR) as well as proliferation induced by either anti‐CD3 or anti‐CD5 mAb. In contrast, anti‐CD5 mAb did not increase proliferation in the AMLR even though both CD5 and WC1 are members of the scavenger receptor cysteine‐rich family of proteins and are expressed by bovine peripheral blood γ/δ T cells. IL‐A29 did not induce proliferation when assessed alone or in the presence of either phorbol myristate acetate (PMA) or interleukin‐2. IL‐A29 also did not induce detectable calcium mobilization when evaluated in the presence of monocytes, PMA, or following cross‐linking of IL‐A29 with anti‐immunoglobulin antibody. We conclude that WC1 is a γ/δ T‐cell lineage‐specific cell‐surface differentiation antigen which is involved in activation of γ/δ T cells using an as yet unidentified pathway.
CD31 (PECAM‐1) is a differentiation antigen lost during human CD4 T‐cell maturation into Th1 or Th2 effector cells
Authors: DEMEURE C. E. , BYUN D. G. , YANG L. P. , VEZZIO N. , DELESPESSE G.
Published: November 2003

The CD31 antigen (PECAM‐1) has been reported to be a stable marker for a human CD4 T‐cell subpopulation unable to produce interleukin‐4 (IL‐4). We show here that CD31 expression is not stable inasmuch as CD4 T‐cell lines and clones derived from cell‐sorted neonatal CD31+ cells lose CD31 upon repetitive cycles of stimulation and IL‐2 expansion. Moreover, various cytokines (IL‐1α, IL‐4, IL‐6, transforming growth factor‐β) fail to reinduce CD31 on CD31− clones. Whereas all CD31+ CD4 T cells rapidly express high levels of the CD45RO antigen and down‐regulate the l‐selectin antigen after priming, CD31 disappears more slowly because only part of the cells lose CD31 expression upon each cycle of stimulation. Loss of CD31 reflects a functional maturation of CD45RO+ cells since, in a system which favours the development of Th2 effectors, IL‐4 is produced by CD31− but not CD31+ effector T cells, whereas interferon‐γ is produced by both types of cells. However, CD31 is not a Th1 marker since it is not expressed on several Th1 antigen‐specific clones. We conclude that CD31 is a maturation marker expressed on the great majority of naive CD45RO− CD4 T cells and on a subset of CD45RO+ CD4 T cells that are at an intermediate stage of maturation.
Protein kinase C isotypes θ, δ and η in human lymphocytes: differential responses to signalling through the T‐cell receptor and phorbol esters
Authors: KEENAN C. , LONG A. , VOLKOV Y. , KELLEHER D.
Published: November 2003

The repertoire of novel and atypical protein kinase C (PKC) isotypes present in human T cells and their subcellular localization have not been fully characterized. We detected calcium‐independent PKC activity in whole cell fractions from unstimulated peripheral blood lymphocytes (PBL). Towards an understanding of the role of PKC isoforms in lymphocyte activation we have studied the expression of calcium‐independent PKC isoforms θ, δ and η in PBL. With isoform‐specific antibodies we detected the presence of PKC θ and δ in whole cell fractions from unstimulated human PBL by Western blot analysis. In addition, immunocytochemical analysis confirmed the presence of the novel PKC isoform PKC η in PBL. Using immunocytochemistry, PKC θ, δ and η had distinct patterns of redistribution following activation by phorbol myristate acetate (PMA). However, signalling through the T‐cell receptor (TCR) did not appear to induce such changes in isoenzyme redistribution. These findings indicate that activation of lymphocytes either through the TCR–CD3 complex or with PMA induces different signalling pathways with respect to calcium‐independent isoenzymes. Signalling through receptors other than the CD3 complex may be involved in activation of these isotypes.
Applications of scanning optical microscopy in materials science to detect bulk microdefects in semiconductors
Authors: Török P. , Mule'Stagno L.
Published: November 2003

We review the application of scanning optical microscopy to bulk microdefect detection in semiconductor materials. After an extensive literature review we summarize theoretical aspects of the scanning infra‐red microscope and describe the theory of contrast formation. We also show experimental examples of scanning infra‐red images taken by different modes of the microscope and give an experimental confirmation of the contrast theory.
Host immune response evasion strategies in Ornithodoros erraticus and O. moubata and their relationship to the development of an antiargasid vaccine
Authors: ASTIGARRAGA AMAIA , OLEAGA‐PÉREZ ANA , PÉREZ‐SÁNCHEZ RICARDO , BARANDA JOSÉ. A. , ENCINAS‐GRANDES ANTONIO
Published: November 2003

We analysed in mice why the salivary gland extract (SGE‐2) from Ornithodoros erraticus and O. moubata induce a protective response with Freund's adjuvants (FAs) in swine while the saliva, in natural conditions, does not. Such protection has been ascribed to the fact that administration of SGE‐2 plus FAs permits the recognition of certain salivary components that under natural conditions are not immunogenic. The present findings confirm this hypothesis since in mice, which are unable to recognize the above components, the SGE‐2‐FAs do not induce any protection. We rule out the possibility that the cause of this could lie in the absence of prostaglandin E2 in the SGE‐2 (vs saliva) since it is not present in either fluid. Neither could it be due to a change in antibody isotype since those induced by parasites bites and by the SGE‐2‐FAs are the same (IgG2a > IgG1 > IgG2b; not IgG3, IgM, IgE). No IgG2a were seen when the SGE‐2 were administered alone or with alum or ricin. It is therefore suggested that first responses would be Th1 and the second ones Th2, although no IgE is seen in the latter responses either. The parasites do not require complement to feed; by contrast, they block its activation and skin cellular infiltrates, such as those elicited by IL‐8, MCP‐1 and C5a, do not affect them, regardless of the presence or not of antitick antibodies.
Anaplasma marginale: effect of the treatment of cattle with an interferon γ‐neutralizing monoclonal antibody or the nitric oxide synthetase inhibitor aminoguanidine on the course of infection
Authors: GALE K.R. , LEATCH G. , DIMMOCK C.M. , WOOD P.R.
Published: November 2003

Cattle undergoing initial infection with the rickettsia Anaplasma marginale were treated with either a monoclonal antibody (MoAb) with neutralizing activity for bovine interferon gamma (IFN‐γ) or aminoguanidine (AG), a specific inhibitor of the inducible form of nitric oxide synthetase (iNOS). Plasma levels of MoAb and AG were measured over the time of administration. The course of A. marginale infection was not altered in the MoAb‐treated cattle relative to untreated controls. In cattle treated with AG however, A. marginale infection was significantly ameliorated, as judged by lower parasite levels and decreased anaemia in these cattle relative to the controls. The implications of these findings in relation to the basis for immunity against this economically important haemoparasite are discussed.
Antiphytoviral activity of 1‐morpholinomethyl‐tetrahydro‐2(1H)‐pyrimidinone (DD13)
Authors: Yordanova A. , Karparov A. , Stoimenova E. , Starcheva M.
Published: November 2003

The antiphytoviral activity of 1‐morpholinomethyl‐tetrahydro‐2(1H)‐pyrimidinone (DD13) in a test system including protoplast cultures, surviving tissues and greenhouse plants was examined. The inhibitory effect was quantitatively investigated by immunofluorescence and enzyme linked immunosorbent assay. The antiviral action in vitro was 96%. The first 6 h after inoculation was the most sensitive period of the tomato mosaic virus (ToMV) reproduction cycle. DD13 possessed a protective effect in 97–100% plants infected with ToMV and cucumber mosaic virus (CMV).
Sources of resistance to Pseudomonas syringae pv. phaseolicola races in Phaseolus vulgaris
Authors: TAYLOR J. D. , TEVERSON D. M. , DAVIS J. H. C.
Published: November 2003

One thousand and forty‐eight Phaseolus bean accessions were evaluated for resistance to six races of Pseudomonas syringae pv. phaseolicola. The accessions originated from regions of the Americas and Africa where the disease is important and included wild type accessions and some known resistance sources. Resistance, graded on a five‐point scale, was of two types: qualitative, which was shown to be race‐specific, and quantitative. Race specific resistance genes (R‐genes) were detected in 49.4% of accessions with the following gene frequencies: R1 (10.3%), R2 (0.3%), R3 (25.0%), R4 (35.0%) and R5 (0.2%).

Evidence for quantitative variation in resistance, in the absence of specific R‐genes, was shown by the distribution of infection scores, 76% of accessions showing maximum susceptibility (grades 4–5), 23% showing intermediate resistance (grades 2–4), and 1% showing high levels of quantitative resistance (grades 1–2). The last 1% of accessions showed interactions which were not race‐specific and it is suggested that they may possess race non‐specific resistance. It is possible that several of the accessions in this category carry the recessive gene derived from PI 150414. Other accessions were of unknown parentage and may represent new sources of quantitative, potentially race non‐specific, resistance. It is suggested that the combination of race specific and race non‐specific resistance could provide an effective strategy for establishing durable resistance.
Biological control of damping‐off of sugar beet by Pseudomonas putida applied to seed pellets
Authors: SHAH‐SMITH D. A. , BURNS R. G.
Published: November 2003

Pseudomonas putida 40RNF applied to seed pellets reduced the occurrence of Pythium damping‐off of sugar beet. A density of 6 × 107 40RNF per pellet reduced Pythium damping‐off from 70 to 26% when seeds were sown in artificially infested soil (250 propagules Pythium ultimum per g dry soil). The efficacy of 40RNF was dependent on its density in the seed pellet (in the range 2 × 104–6 × 108 per pellet) and on the number of propagules of Pythium in soil. 40RNF declined to or stabilized at approximately 1 × 106 per pellet 3 days after planting, and this was independent of the inoculum density. This indicated that the crucial steps resulting in damping‐off of sugar beet caused by Pythium ultimum must occur within 3–4 days of sowing. 40RNF reduced pericarp colonization by P. ultimum by 43% 48 h after planting and caused a 68% decrease in the number of sporangia of P. ultimum in the surrounding soil (0.0–5.0 mm). P. putida 40RNF also reduced pre and post‐emergence damping‐off (from 69.5 to 37.5%) caused by indigenous populations of Pythium species in an infested soil and this was as effective as the fungicide hymexazol (69.5 to 40%).
Histaminergic Regulation of NK‐Cells: Protection Against Monocyte‐Induced Apoptosis
Authors: HANSSON M. , ASEA A. , HERMODSSON S. , HELLSTRAND K.
Published: November 2003

Human natural killer (NK) cells (with CD3−/56+ phenotype) acquired features characteristic of apoptosis after incubation with autologous monocytes, as revealed by apoptotic nuclear morphology and degradation of DNA into oligonucleosomal fragments. The monocyte‐induced apoptosis in NK‐cells was prevented by the biogenic amine histamine at concentrations exceeding 0.1 μM. The protective effect of histamine was blocked by the H2‐receptor (H2R) antagonist ranitidine but not by AH202399 A, a chemical control to ranitidine devoid of H2R affinity. It is concluded that histaminergic mechanisms may serve to protect NK cells from damage inflicted by products of the oxidative metabolism of monocytes.
Activation of the Complement, Coagulation, Fibrinolytic and Kallikrein–Kinin Systems During Attacks of Hereditary Angioedema
Authors: WAAGE NIELSEN E. , THIDEMANN JOHANSEN H. , HØGÄSEN K. , WUILLEMIN W. , HACK C. E. , MOLLNES T. E.
Published: November 2003

Five patients with hereditary angioedema (HAE) were studied during attacks and remission as were healthy controls. The high levels of C1/C1‐INH complexes, low C4 and high ratio C4 activation products (C4bc)/C4 also differed significantly during remission compared to controls.During attacks C4bc/C4 increased (922–2007; P=0.022, remission versus attacks, median values throughout), C2 and CH50 dropped (111–31%; P=0.043 and 110–36%; P=0.016, respectively), TCC (C5b‐9) increased (0.88–1.23 AU/ml; P=0.028). Cleavage of HK increased to be almost complete during attacks (20–90%; P=0.009). While factor XIa/serpin‐complexes did not increase, a more than twofold rise in thrombin/antithrombin‐complexes (0.20–0.50 μg/l; P=0.009) and in plasmin/alpha‐2‐antiplasmin‐complexes (7.3–17 nmol/l; P=0.028) was observed. For the first time cascade activation in HAE was studied simultaneously, and corroborates that attacks lead to activation of the kallikrein‐kinin system, fibrinolysis and early part of the classical complement pathway. In addition, the authors present novel data of terminal complement and coagulation activation, the latter apparently not via FXIa.
High Dose IL‐2‐Activated Murine Natural Killer (A‐NK) Cells Accumulate Glycogen and Granules, Lose Cytotoxicity, and Alter Target Cell Interaction In Vitro
Authors: UNGER M. L. , HOKLAND M. , BASSE P. H. , NANNMARK U. , JOHANSSON B. R.
Published: November 2003

Activated natural killer (A‐NK) cells, defined by immunophenotype and selected by adherence to the plastic, were cultured from murine splenocytes for up to 10 days with the addition of 1000 U/ml of recombinant human IL‐2 at 48 h intervals. During culture days 2–4 with high DNA synthesis the initially non‐granulated small cells established large granular lymphocyte (LGL) morphology and then differentiated further into giant hypergranulated cells with huge accumulations of glycogen. Timed EM observations indicated that specific dual‐compartment (lytic) granules arose by a sequence of events starting with neo‐synthesis of small progenitors with a dense core and a few membranous lamellae at one pole. Core and vesicular regions probably expanded independently to give the mature organization of the granule. Eventually, the vesicular region of granules contained large amounts of multi‐lamellar material and probable debris, and the dense core could be multiplied. Intracellular proteoglycans, visualized with Cupromeronic Blue cytochemistry, were organized in a three‐dimensional network within the dense cores. In contrast with earlier reports, and in spite of several‐fold increased granularity, the in vitro cytotoxicity of the A‐NK cells against YAC‐1 and B16 cells decreased after the third day of culture. A‐NK cells with glycogen accumulations caused focal clearing in melanoma monolayers whereas younger effectors adhered to the targets. It is concluded that high dose IL‐2 stimulation causes more far‐going progressive morphological and functional differentiations of the A‐NK cells than has previously been observed with bearing for the use of these cells in experimental adoptive immunotherapy.
Arthritis Susceptibility in Mice Expressing Human Type II Collagen in Cartilage
Authors: MALMSTRÖM V. , HO K. K. Y. , LUN J. , TAM P. P. L. , CHEAH K. S. E. , HOLMDAHL R.
Published: November 2003

Collagen type II (CII) induced arthritis (CIA) in mice is an experimental model for rheumatoid arthritis. Induction with non‐self (e.g. human) CII induces severe arthritis whereas the mice are less susceptible to induction with self CII (i.e. mouse). To analyse whether an autoimmune response to human CII can develop and is pathogenic the authors have established transgenic mice expressing human CII in cartilage and backcrossed them into two different gene backgrounds susceptible to CIA (DBA/1 and C3H.Q). The transgenic human CII expression was restricted to cartilage and did not disturb cartilage morphology or lead to chondrodystrophy. In addition, development of stress‐induced arthritis was not affected by the transgene. The cartilage specific expression of human CII reduced, but did not eliminate, the susceptibility to CIA irrespective of the species source (human, bovine, chick, rat) of CII used for immunization. A common denominator between these heterologous CII in comparison with mouse CII is the previously defined CII 256–270 epitope. An expression level dependent T‐cell tolerance was seen in this epitope as well as to the entire CII. However, all human transgenic mouse lines could still mount significant autoreactive T‐ and B‐cell responses. Approximately 10% of the transgenic mice developed arthritis after immunization with human CII. These findings show, therefore, that cartilage‐located human CII induce tolerance but can nevertheless be a target for development of arthritis.
Conversion of In Vitro Cultured Human Monocytes into Effective Presenters of an HER2/neu‐Encoded CTL Peptide Epitope
Authors: WASSERMAN K. , CORSI M. M. , SZEKELY L. , KONO K. , MAES H. H. , KIESSLING R.
Published: November 2003

Tumour‐derived peptides have been surveyed, in a variety of systems, for their ability to elicit cytokine release from class I restricted T cells. Analogous studies on ovarian carcinoma have employed the antigen‐processing defective T2 cell line. Purified dendritic cells (DC) have been reported to act as highly effective APC. A facile method was developed whereby DC‐like cells were generated from monocyte precursors. Herein, evidence is presented suggesting DC‐like cells are superior to T2 with respect to their ability to present a defined CTL epitope associated with ovarian carcinoma.
IL‐10 Plays a Role in the Modulation of Human Granulomatous Hypersensitivity Against Schistosoma mansoni Eggs Induced by Immune Complexes
Authors: REZENDE S. A. , SILVA‐TEIXEIRA D. N. , DRUMMOND S. C. , GOES A. M.
Published: November 2003

It has been demonstrated that the chronic intestinal form of schistosomiasis is associated with the establishment and maintenance of a variety of immunoregulatory mechanisms that lead to a diminished granulomatous reaction to parasite eggs. Using an in vitro model of granuloma reaction we showed that immune complexes (IC) isolated from the sera of chronic intestinal schistosomiasis patients were able to reduce the granulomatous hypersensitivity (developed by peripheral blood mononuclear cells (PBMC) from schistosomiasis patients) to soluble egg antigen (SEA)‐conjugated polyacrylamide beads (PB–SEA). This inhibitory activity, mediated by IC, was also observed in the proliferative response of PBMC stimulated with SEA and soluble worm antigen preparation (SWAP). Furthermore, we observed a decrease in TNF‐α and an increase in IL‐10 production by PBMC treated with IC in an in vitro granuloma reaction. This phenomenon was also seen in a cell proliferation assay when PBMC were treated with IC and stimulated with S. mansoni antigens. These results demonstrate that circulating IC may down‐regulate PBMC reactivity to S. mansoni antigens by changing the cytokine pattern produced by these cells.
Quality assurance of clinical transfusion practice by implementation of the privilege of blood prescription and computerized prospective audit of blood requests
Authors: Marconi M. , Almini D. , Pizzi M. N. , Riccardi D. , Bergamaschi W. , Giovanetti A. M. , Rebulla P. , Sirchia G.
Published: November 2003

Guidelines, algorithms and recommendations have been issued in the attempt to ensure appropriateness of transfusion practice, but the results are less than satisfactory, mainly due to the difficulty to turn paper procedures into actual practice. In our hospital we have tried to overcome this difficulty through the implementation of a quality assurance programme which includes giving the privilege of nonurgent blood prescription to a limited number of physicians and a computerized prospective audit of blood requests. The latter is performed through verification of the compliance of blood requests, which are designed to include a patient's laboratory and clinical data, with hospital guidelines for the proper use of blood.  In the 12 months since implementation of the computerized prospective audit the transfusion service has evaluated 7884 requests. Of these, 63.4% (n = 4998) were for red blood cells, 21.1% (n = 1664) for platelets and 15.5% (n = 1222) for fresh frozen plasma. The prospective audit showed that 96.8% and 98.1% of requests for red units and platelets were appropriate, respectively. Conversely, approximately 27% of plasma requests did not comply with guidelines, mainly because the evidence of coagulopathy was missing. However, inappropriateness of plasma requests for elective general surgery decreased from 39% at the onset of the programme to 14% in the last trimester considered. Moreover, the evaluation by retrospective audit of the proportion of patients transfused with both red blood cells and plasma in the perioperative period out of those transfused with red blood cells only, as an indicator of unwanted reconstitution of whole blood, showed that this proportion decreased from 47.6% (320/672) in the 12 months before implementation of computerized audit to 37.8% (244/646) in the following 12 months (difference = −9.8%, 95% confidence interval of the difference from −4.5% to −15.1%; P < 0.005 by χ2 test). Our initial experience, together with the present system, shows that (1) the restriction of nonurgent blood prescription to a group of clinicians more educated in transfusion medicine than average clinicians practicing in a large multispecialty hospital is feasible; (2) prospective audit is a useful tool for assuring the quality of blood requesting.
Provider‐sponsored virtual communities for chronic patients: improving health outcomes through organizational patient‐centred knowledge management
Authors: Winkelman Warren J. , Choo Chun Wei
Published: November 2003

Patients with long‐term chronic disease experience numerous illness patterns and disease trends over time, resulting in different sets of knowledge needs than patients who intermittently seek medical care for acute or short‐term problems. Health‐care organizations can promote knowledge creation and utilization by chronic patients through the introduction of a virtual, private, disease‐specific patient community. This virtual socialization alters the role of chronic disease patients from external consumers of health‐care services to a ‘community of practice’ of internal customers so that, with the tacit support of their health‐care organization, they have a forum supporting the integration of knowledge gained from the experiences of living with chronic disease in their self‐management. Patient‐centred health‐care organizations can employ the virtual community to direct and support the empowerment of chronic patients in their care.
Characterization of a defensin in bark and fruit tissues of peach and antimicrobial activity of a recombinant defensin in the yeast, Pichia pastoris
Authors: Wisniewski Michael E. , Bassett Carole L. , Artlip Timothy S. , Webb Robert P. , Janisiewicz Wojciech J. , Norelli John L. , Goldway Martin , Droby Samir
Published: November 2003

Herein the cloning and characterization of a defensin gene (PpDfn1) from a cDNA library made from peach (Prunus persica[L.] Batsch) winter bark tissues is described. A partial clone obtained from the library was extended to full length by 5′ Rapid Amplification of cDNA Ends (RACE). The open reading frame of 237 bp codes for a 79 amino acid peptide related to the defensin family of proteins. Sequence comparison of the encoded protein using blast analysis revealed significant homology to defensins from other plant species. RNA gel blot analysis indicated that the gene is seasonally expressed in bark tissues of 1‐year‐old shoots, and is also expressed in early fruit development. Results of quantitative RT‐PCR and protein blot analysis were similar to those of RNA gel blot analyses for the bark tissues. A recombinant version, rDFN1 was expressed in the yeast, Pichia pastoris. It was found that rDFN1 inhibited germination of the fungal pathogens Penicillium expansum and Botrytis cinerea, but not the Gram‐negative bacterium Erwinia amylovora. The potential physiological role of PpDFN1 and its antimicrobial properties are discussed.