bluuebunny/crossref_metadata_2025 huggingface dataset:
Adenomatous polyposis coli and EB1 localize in close proximity of the mother centriole and EB1 is a functional component of centrosomes
Authors: Louie Ryan K. , Bahmanyar Shirin , Siemers Kathleen A. , Votin Violet , Chang Paul , Stearns Tim , Nelson W. James , Barth Angela I. M.
Published: February 2004

Adenomatous polyposis coli (APC) and End-binding protein 1 (EB1) localize to centrosomes independently of cytoplasmic microtubules (MTs) and purify with centrosomes from mammalian cell lines. Localization of EB1 to centrosomes is independent of its MT binding domain and is mediated by its C-terminus. Both APC and EB1 preferentially localize to the mother centriole and EB1 forms a cap at the end of the mother centriole that contains the subdistal appendages as defined by ϵ-tubulin localization. Like endogenous APC and EB1, fluorescent protein fusions of APC and EB1 localize preferentially to the mother centriole. Depletion of EB1 by RNA interference reduces MT minus-end anchoring at centrosomes and delays MT regrowth from centrosomes. In summary, our data indicate that APC and EB1 are functional components of mammalian centrosomes and that EB1 is important for anchoring cytoplasmic MT minus ends to the subdistal appendages of the mother centriole.
Involvement of the cohesin Rad21 and SCP3 in monopolar attachment of sister kinetochores during mouse meiosis I
Authors: Parra María Teresa , Viera Alberto , Gómez Rocío , Page Jesús , Benavente Ricardo , Santos Juan Luis , Rufas Julio S. , Suja José A.
Published: February 2004

SCP3 is a meiosis-specific structural protein appearing at axial elements and lateral elements of the synaptonemal complex. We have analysed the behaviour of SCP3 and the cohesin subunit Rad21 in mouse spermatocytes by means of a squashing technique. Our results demonstrate that both proteins colocalize and are partially released from chromosome arms during late prophase I stages, although they persist at the interchromatid domain of metaphase I bivalents. Thus, Rad21 cannot be considered a `mitotic'-specific variant, but coexists with Rec8. During late prophase I SCP3 and Rad21 accumulate at centromeres, and together with the chromosomal passenger proteins INCENP and aurora-B kinase, show a complex `double cornet'-like distribution at the inner domain of metaphase I centromeres beneath the associated sister kinetochores. We have observed that Rad21 and SCP3 are displaced from centromeres during telophase I when sister kinetochores separate, and are not present at metaphase II centromeres. Thus, we hypothesise that Rad21, and the superimposed SCP3 and SCP2, are involved in the monopolar attachment of sister kinetochores during meiosis I, and are not responsible for the maintenance of sister-chromatid centromere cohesion during meiosis II as previously suggested.
The 12 kDa FK506-binding protein, FKBP12, modulates the Ca2+-flux properties of the type-3 ryanodine receptor
Authors: Van Acker Kristel , Bultynck Geert , Rossi Daniela , Sorrentino Vincenzo , Boens Noel , Missiaen Ludwig , De Smedt Humbert , Parys Jan B. , Callewaert Geert
Published: February 2004

We have characterised the functional regulation of the type-3 ryanodine receptor by the 12 kDa FK506-binding protein. Wild-type type-3 ryanodine receptor and mutant type-3 ryanodine receptor in which the critical valine at position 2322 in the central 12 kDa FK506-binding protein binding site was substituted by aspartate, were stably expressed in human embryonic kidney cells. In contrast to the wild-type receptor, the mutant receptor was strongly impaired in binding to immobilised glutathione S-transferase 12 kDa FK506-binding protein. Caffeine-induced 45Ca2+-efflux was markedly increased in cells expressing mutant type-3 ryanodine receptor whereas the maximal-releasable Ca2+ was not affected. Confocal Ca2+ imaging provided clear evidence for a much higher sensitivity of the mutant receptor, which showed global Ca2+ release at about 20-fold lower caffeine concentrations than the wild-type receptor. Spontaneous Ca2+ sparks were observed in both wild-type- and mutant-expressing cells but the number of sparking cells was about 1.5-fold higher in the mutant group, suggesting that the degree of FK506 binding controls the stability of the closed state of ryanodine receptor channels. Furthermore, overexpression of 12 kDa FK506-binding protein decreased the number of sparking cells in the wild-type-expressing cells whereas it did not affect the number of sparking cells in cells expressing the mutant receptor. Concerning spark properties, the amplitude and duration of Ca2+ sparks mediated by mutant channels were significantly reduced in comparison to wild-type channels. This suggests that functional coupling between different mutant type-3 ryanodine receptor channels in a cluster is impaired. Our findings show for the first time that the central binding site for the 12 kDa FK506-binding protein of type-3 ryanodine receptor, encompassing the critical valine proline motif, plays a crucial role in the modulation of the Ca2+ release properties of the type-3 ryanodine receptor channel, including the regulation of both global Ca2+ responses and spontaneous Ca2+ sparks.
The bHLH TAL-1/SCL regulates endothelial cell migration and morphogenesis
Authors: Lazrak Monia , Deleuze Virginie , Noel Danièle , Haouzi Delphine , Chalhoub Elias , Dohet Christiane , Robbins Ian , Mathieu Danièle
Published: February 2004

The basic helix-loop-helix tal-1 gene (or scl), known for its fundamental role in embryonic and adult hematopoiesis in vertebrates, is also required for embryonic vascular remodeling. In adults, TAL-1 protein is undetectable in quiescent endothelium but it is present in newly formed vessels including tumoral vasculature, indicating its involvement in angiogenesis. Here, we demonstrate that TAL-1 expression is tightly regulated during in vitro angiogenesis: it is low during the initial step of migration and is upregulated during formation of capillary-like structures. We investigated whether ectopic expression of either wild-type TAL-1 or a dominant-negative mutant lacking the DNA-binding domain (Δ-bas) modulates the activity of human primary endothelial cells in the angiogenic processes of migration, proliferation and cell morphogenesis. Overexpression of either wild-type or Δ-bas TAL-1 affected chemotactic migration of primary endothelial cells without modifying their proliferative properties. Ectopic expression of wild-type TAL-1 accelerated the formation of capillary-like structures in vitro and, in vivo, enhanced vascularisation in mice (Matrigel implants) associated with a general enlargement of capillary lumens. Importantly, transduction of the mutant Δbas completely impaired in vitro angiogenesis and strongly inhibited vascularisation in mice. Taken together, our data show that TAL-1 modulates the angiogenic response of endothelial cells by stimulating cell morphogenesis and by influencing their behavior in migration. This study highlights the importance of TAL-1 regulation in postnatal vascular remodeling and provides the first physiological evidence that links TAL-1 activity to endothelial cell morphogenic processes.
An investigation of first‐order false belief understanding of children with congenital profound visual impairment
Authors: Green Sarah , Pring Linda , Swettenham John
Published: February 2004

This study assessed theory of mind understanding in children with congenital profound visual impairment (CPVI): children who have had no access to visual information throughout development. Participants were 18 children with CPVI and no other impairments, aged between 5 and 11 years, and 18 children with normal vision, matched individually on chronological age, verbal IQ and verbal mental age. Three first‐order false belief tasks were presented twice each; the three tasks varied in the extent of deception and involvement of the child. Six of the children with CPVI failed one or more of the false belief tasks; all sighted children passed all of the tasks. The manipulations of deception and involvement did not influence the performance of the children with CPVI. Participant characteristics of the children with CPVI were examined in relation to their performance on the false belief tasks: chronological age and type of school attended were not found to be related to performance; verbal IQ and verbal mental age were found to differ in children with good and poor performance on the false belief tasks. The results are consistent with either a general pattern of delay in theory of mind development for children with CPVI, or with a subset of children who have longer‐term difficulties in this area.
Bullying and social identity: The effects of group norms and distinctiveness threat on attitudes towards bullying
Authors: Ojala Kris , Nesdale Drew
Published: February 2004

Drawing from social identity theory (Tajfel & Turner, 1979), an experiment was carried out to determine the extent to which children's attitudes towards bullying could be moderated by in‐group norms and perceived threat to group distinctiveness. The study investigated the responses of 120 male primary school students aged 10‐13 years from five schools. The children read a story about a popular in‐group and an unpopular out‐group which involved the manipulation of three variables: the norms of the in‐group (bullying vs. fairness); distinctiveness threat (out‐group similarity vs. out‐group difference); and the behaviour of the in‐group character towards the out‐group character (bullying vs. helpful). It was predicted that a perceived threat to group distinctiveness, represented by similarity between the in‐group and the out‐group, and salient group norms that prescribed either bullying or fairness, would moderate the acceptability of bullying behaviours. Two story response measures were analysed: in‐group character liking and whether the in‐group character would be retained as a group member following his behaviour. The strongest support for social identity theory was revealed in the retention of in‐group character variable. The in‐group character was much more likely to have been retained as a group member when he behaved in accordance with group norms. Evidence was also found that bullying was more acceptable when directed at an out‐group member who was similar and therefore possibly represented a threat to the in‐group.
Exploring the coherence of young children's explanatory abilities: Evidence from generating counterfactuals
Authors: Sobel David M.
Published: February 2004

Researchers who advocate the hypothesis that cognitive development is akin to theory formation have also suggested that young children possess distinct systems for explaining physical, psychological, and biological principles (see, e.g., Wellman & Gelman, 1992). One way this has been investigated is by examining how children explain human action: Children explain intentional and accidental actions by appealing to psychological principles, and explain impossible physical or biological action in terms of the underlying principles of those domains (Schult & Wellman, 1997). The current investigation examined the coherence of children's explanatory systems by eliciting explanations of possible and impossible physical, psychological, and biological events. Then, in a separate set of stories, children were asked to generate counterfactual alternatives for characters who wanted to perform an event, but did not, either because of a mishap or because the event was impossible. Overall, children were better at generating explanations for why events were impossible than recognizing that no alternative could be generated for impossible events. However, there was some evidence that children's explanatory abilities predicted whether they could correctly reject cases where no counterfactual alternative could be generated. The results lend support to the hypothesis that children's causal knowledge is coherently organized in domain‐specific knowledge structures.
The role of the physical context in supporting young children's use of spatiotemporal organization in recall
Authors: Kannass Kathleen N. , Plumert Jodie M. , McDermott Jessica , Moore Bethany , Durich Nathan
Published: February 2004

Four experiments were conducted to investigate the role of the physical context in supporting 3‐ to 5‐year‐olds' use of spatiotemporal organization in recall. Children were familiarized with several target items and their corresponding landmarks arranged along a path in a model park. After familiarization, an experimenter removed the target items from the park. In Experiment 1, 3‐ and 4‐year‐olds recalled the missing items with the park either in view or out of view. When the park model was in view, 4‐ year‐olds used the order of the items along the path to structure their recall. In Experiment 2, 4‐ and 5‐year‐olds recalled the missing items with the landmarks arranged either in the same order as in familiarization or in a new order. Children used the order of landmarks along the path at test to structure their recall, even though the order of landmarks changed from familiarization to test. Experiment 3 was identical to Experiment 2, except that the path was removed from the park. Five‐year‐olds used the order of landmarks along the path at test to structure their recall when the order of landmarks remained the same from familiarization to test, but had much more difficulty doing so when the order of landmarks changed from familiarization to test. Using a more difficult task, Experiment 4 revealed that spatiotemporal organization was positively related to amount recalled. Together, these findings suggest that the structure of the physical environment plays an important role in supporting young children's use of spatiotemporal organization in recall.
Discovery and maintenance of the many‐to‐one counting strategy in 4‐year‐olds: A microgenetic study
Authors: Blöte Anke W. , Van Otterloo Sandra G. , Stevenson Claire E. , Veenman Marcel V. J.
Published: February 2004

This study investigated the development of the many‐to‐one counting strategy in 4‐ year‐old children. In the first experiment, 52 children participated. Their development with respect to two kinds of tasks, a hidden‐items task and a needed‐items task, was studied over four sessions. Children (n = 28) who accurately used the many‐to‐one strategy in Session 4 also participated in the second experiment. These children were presented with more difficult hidden‐ and needed‐items tasks. It was found that children often produced the strategy for the first time on tasks with relatively few items. Most children then kept producing it, even if they initially did not obtain much profit from its use because of counting errors. Increasing task difficulty resulted in children making more counting errors or reverting to invalid strategies depending on the nature of the new task.
Magical thinking in judgments of causation: Can anomalous phenomena affect ontological causal beliefs in children and adults?
Authors: Subbotsky Eugene
Published: February 2004

In four experiments, 4‐, 5‐, 6‐ and 9‐year‐old children and adults were tested on the entrenchment of their magical beliefs and their beliefs in the universal power of physical causality. In Experiment 1, even 4‐year‐olds showed some understanding of the difference between ordinary and anomalous (magical) causal events, but only 6‐year‐olds and older participants denied that magic could occur in real life. When shown an anomalous causal event (a transformation of a physical object in an apparently empty box after a magic spell was cast on the box), 4‐ and 6‐year‐olds accepted magical explanations of the event, whereas 9‐year‐olds and adults did not. In Experiment 2, the same patterns of behaviour as above were shown by 6‐ and 9‐year‐olds who demonstrated an understanding of the difference between genuine magical events and similarly looking tricks. Testing the entrenchment of magical beliefs in this experiment showed that 5‐year‐olds tended to retain their magical explanations of the anomalous event, even after the mechanism of the trick had been explained to them, whereas 6‐ and 9‐year‐olds did not. In Experiment 3, adult participants refused to accept magical explanations of the anomalous event and interpreted it as a trick or an illusion, even after this event was repeated 4 times. Yet, when in Experiment 4 similar anomalous causal events were demonstrated without reference to magic, most adults acknowledged, both in their verbal judgments and in their actions, that the anomalous effects were not a fiction but had really occurred. The data of this study suggest that in the modern industrialized world, magical beliefs persist but are disguised to fit the dominant scientific paradigm.
Two-Dimensional Mapping of Copper and Zinc in Liver Sections by Laser Ablation–Inductively Coupled Plasma Mass Spectrometry
Authors: Kindness Andrew , Sekaran Chandra N , Feldmann Jörg
Published: February 2004

Background: Metals are not homogeneously distributed in organ tissues. Although most mapping techniques, such as histologic staining methods, have been developed for element imaging on a subcellular level, many suffer from either low precision or poor detection limits. Therefore, small variations in elemental distribution cannot be identified. We developed a method for two-dimensional mapping of trace elements to identify the influence of metabolic zonation by the liver on trace element distribution.

Methods: A prepared homogeneous Certified Reference Material (CRM; LGC 7112, pig liver) was used to characterize the laser ablation–inductively coupled plasma time-of-flight mass spectrometry (LA-ICP-MS) in terms of precision. Different isotopes for copper and zinc were monitored, and the use of carbon as an internal standard was investigated to correct for differences in ablation efficiency to identify the most precise mapping technique for liver samples.

Results: For the homogeneous CRM, the reproducibility of the copper and zinc signals was ∼3–24% depending on spot size and number of pulses. When carbon was used as an internal standard, the reproducibility was improved significantly. Line scan signals over a length of 1.5 mm were more precise [relative SD (RSD), 1.6–6.1% for copper (63Cu, 65Cu) and zinc (64Zn, 66Zn) depending on the spot size, the scanning speed, and the element]. Thin section of sheep liver achieved precisions of 27–59% (raster scan) and 9–47% (line scan) RSD for copper, whereas the precision for zinc was significantly better: 8–18% (raster scan) and 4–21% (line scan) RSD. Long line scans and two-dimensional element maps of the thin sections revealed the zonation of copper in sheep liver containing extremely low copper concentrations.

Conclusion: Elemental mapping of trace elements generated by LA-ICP-MS can be very precise so that small changes in the elemental concentration in the tissue can be detected and nonuniform spatial distribution of the elements in tissues can be established.
Apolipoprotein C-III Isofocusing in the Diagnosis of Genetic Defects in O-Glycan Biosynthesis
Authors: Wopereis Suzan , Grünewald Stephanie , Morava Éva , Penzien Johannes M , Briones Paz , Garcı́a-Silva M Teresa , Demacker Pierre N M , Huijben Karin M L C , Wevers Ron A
Published: February 2004

Background: Defects in the biosynthesis of N-glycans may be found by isoelectric focusing (IEF) of plasma transferrin. No test is available to demonstrate O-glycan biosynthesis defects.

Methods: We used isoforms of apolipoprotein C-III (apoC-III) as a marker for the biosynthesis of core 1 mucin type O-glycans. Plasma samples from patients with primary defects and secondary alterations in N-glycan biosynthesis were studied by apoC-III isofocusing.

Results: Age-related reference values for apoC-III were determined. Plasma samples from patients with the primary congenital disorders of glycosylation (CDG) types Ia–Ic, Ie, If, IIa, and IId all showed a normal apoC-III isofocusing profile. Plasma from two patients with CDG type IIx were tested: one showed a normal apoC-III distribution, whereas the other showed a hypoglycosylation profile. In plasma from patients with hemolytic uremic syndrome (HUS), a hypoglycosylation profile was obtained.

Conclusions: IEF of apoC-III is a rapid and simple technique that may be used as a screening assay for abnormalities in core 1 mucin type O-glycans. Evidence that a patient in this study has a primary genetic defect affecting both N- and O-glycosylation provides the first example of an inborn error of metabolism affecting the biosynthesis of core 1 mucin type O-glycans. Our data narrow the options for the position of the primary defect in this patient down to a step in the biosynthesis, activation, or transfer of galactose or N-acetylneuraminic acid to both N- and O-glycans. Circulating neuraminidase excreted by Streptococcus pneumoniae caused the high percentage of asialo apoC-III in two HUS patients.
Rapid Separation of LDL Subclasses by Iodixanol Gradient Ultracentrifugation
Authors: Davies Ian G , Graham John M , Griffin Bruce A
Published: February 2004

Background: A predominance of small, dense LDL (sdLDL) confers in excess of a threefold increase in coronary heart disease (CHD) risk. The conventional method for the detection of sdLDL, salt density gradient ultracentrifugation (DGUC) has been superseded by more rapid techniques. This report presents novel methodology for the separation of sdLDL by a combination of iodixanol density gradient centrifugation and digital photography.

Methods: LDL subclasses were separated in 3 h from prestained plasma on a self-forming density gradient of iodixanol. LDL subclass profiles were generated by digital photography and gel-scan software. Plasma samples from 106 normo- and dyslipidemic individuals were used to optimize the gradient for the resolution of LDL heterogeneity. A subgroup of 47 LDL profiles were then compared with LDL subclasses separated by salt DGUC.

Results: The peak density of the predominant LDL band correlated significantly with the relative abundance (as a percentage) of sdLDL as resolved by salt DGUC (P <0.001). As shown previously, LDL isolated at a lighter density in iodixanol compared with salt gradients. A predominance of sdLDL corresponded to a peak density on iodixanol of 1.028 kg/L. This density and the area under the LDL profile lying above this density were sensitive and specific markers for the prediction of a predominance of sdLDL (P <0.001) and showed predictable associations with plasma triglycerides (r = 0.59; P <0.001) and HDL (r = −0.4; P <0.001).

Conclusions: This simple method for the detection of sdLDL can differentiate a predominance of sdLDL, is highly reproducible, and can be used preparatively to isolate sdLDL.
Sensitive HPLC-Fluorescence Method for Irinotecan and Four Major Metabolites in Human Plasma and Saliva: Application to Pharmacokinetic Studies
Authors: Poujol Sylvain , Pinguet Frédéric , Malosse Françoise , Astre Cécile , Ychou Marc , Culine Stéphane , Bressolle Françoise
Published: February 2004

Background: We developed gradient HPLC methods for quantification of the antimitotic drug irinotecan (CPT-11) and its four metabolites, SN-38, SN-38 G, 7-ethyl-10-[4-N-(5-aminopentanoic acid)-1-piperidino]-carbonyloxycamptothecin (APC), and 7-ethyl-10-[4amino-1-piperidino]-carbonyloxycamptothecin (NPC), as the sum of the lactone and carboxylate forms, in human plasma and saliva. Camptothecin was used as internal standard.

Methods: The sample pretreatment involved protein precipitation with methanol–acetonitrile (50:50 by volume) followed by acidification with hydrochloric acid to convert the lactone ring-opened form into its lactone form, quantitatively. HPLC separation was performed on a Xterra RP18 column. The excitation wavelength was 370 nm, and the emission wavelength was set at 470 nm for the first 24 min and then at 534 nm for the next 4 min. The stabilities of irinotecan and its four metabolites in plasma, saliva, and acidic extracts were also investigated under various conditions.

Results: Assays were linear in the tested range of 0.5–1000 μg/L. For the five analytes, limits of quantification were 0.5 μg/L in both matrices. The interassay imprecision (as relative standard deviation) was 3.2–14% in plasma and 2.6–5.6% in saliva. Assay recoveries ranged from 92.8% to 111.2% for plasma and 100.1% to 104.1% for saliva. Mean extraction recovery from plasma or saliva was 90%.

Conclusion: The developed assay can be used to determine pharmacokinetic parameters for CPT-11, SN-38, SN-38 G, APC, and NPC in plasma and saliva from patients with metastatic colorectal cancer.
Plasma Fluorescence Scanning and Fecal Porphyrin Analysis for the Diagnosis of Variegate Porphyria: Precise Determination of Sensitivity and Specificity with Detection of Protoporphyrinogen Oxidase Mutations as a Reference Standard
Authors: Hift Richard J , Davidson Brandon P , van der Hooft Cornelis , Meissner Doreen M , Meissner Peter N
Published: February 2004

Background: Variegate porphyria (VP) is the autosomal dominant disorder associated with deficiency of the enzyme protoporphyrinogen oxidase (PPOX). Plasma fluorescence scanning has been reported to be a more sensitive test for VP than traditional fecal chromatography. Previous comparisons of these techniques predated identification of the PPOX gene. We assessed these techniques in a large group of patients characterized for VP at the DNA level.

Methods: We evaluated all patients for whom the genotype and a plasma scan or fecal porphyrin result were available. Mutations were detected by restriction digest analysis. Plasma fluorescence scanning was conducted according to published methods. Fecal porphyrins were identified and quantified by thin-layer chromatography.

Results: Plasma fluorescence scanning was assessed in 679 patients (205 with VP who were carriers of a PPOX mutation, either with disease symptoms or asymptomatic) and fecal analysis in 473 (190 with VP). Sensitivity and specificity of both tests were higher in adults than in children and higher for adults with disease symptoms than for asymptomatic carriers. In a direct comparison in 168 adults (73 with VP), plasma scanning was significantly more sensitive than fecal porphyrin analysis [sensitivity, 0.96 (95% confidence interval, 0.89–0.99) vs 0.77 (0.66–0.85)]. Fecal coproporphyrin [area under the curve, 0.87 (0.83–0.90)] was a better predictor of VP than protoporphyrin [0.80 (0.76–0.84)].

Conclusions: Plasma scanning is a more sensitive and specific test for VP than fecal porphyrin analysis. Neither test is sensitive in children, and both are less sensitive in asymptomatic carriers than in symptomatic cases. DNA analysis therefore remains the preferred method for the identification of carriers, particularly in children.
The place of physiotherapy in the management of pelvic floor dysfunction
Authors: Laycock Jo , Holmes David M
Published: February 2004

Pelvic floor muscle exercises are the mainstay of physiotherapy for urinary incontinence and can be taught by all health professionals. If the patient fails to respond to a three‐month course of pelvic floor exercises, or if she cannot perform a voluntary contraction of the pelvic floor muscles, biofeedback, electrical stimulation and vaginal cone therapy may be appropriate. This review discusses the evidence supporting the use of physiotherapy in the treatment of urinary incontinence. It also reports on the prevalence of the different types of urinary incontinence and describes the need for vaginal examination to determine the contractility of the pelvic floor muscles, including the PERFECT scheme.
Hyperemesis gravidarum
Authors: Neill Anne‐Marie C , Nelson‐Piercy Catherine
Published: February 2004

Nausea and vomiting in early pregnancy are common; however, in some cases symptoms can persist, leading to maternal weight loss. This condition is referred to as hyperemesis gravidarum (HG). Treatment requires recognition, admission to hospital and appropriate rehydration. All practitioners, including those managing low‐risk pregnancies, should be aware of the complications associated with HG. To avoid substandard care a protocol for management should be available in all obstetric units.
Building sustainable partnerships in user involvement
Authors: Fletcher Gillian
Published: February 2004

In a BMJ article published on June 14th 2003, Richard Smith talks of ‘helping doctors prepare for a world where true partnership with patients is the norm’.1 The inexorable move towards user or patient and public involvement (PPI) requires a review of some of the complex issues involved in these changing relationships. Much has been written about PPI over the years, but what do the various terms mean? This article offers an exploration of different types and levels of involvement and some examples of good practice within the maternity and cancer services.
On‐the‐job training: ward and clinic
Authors: MacDougall Jane
Published: February 2004

Calman reforms and the European Working Time Directive have led to a reduction in trainees' hours and length of training. There is an increasing need to provide focused and high quality training, yet this is difficult given the increasing demands on consultant time. Young doctors also need to be encouraged to become self‐directed learners who work well in multidisciplinary teams. This article aims to explore different ways of training on wards and in clinics and suggests how trainers can help trainees to take advantage of the many opportunities for learning available in these settings.
Selective salpingography and tubal catheterisation in the diagnosis and treatment of proximal tubal blockage
Authors: Papaioannou Spyros , Afnan Masoud , Sharif Khaldoun
Published: February 2004

Proximal tubal blockage arises from several pathological processes and is often a sole cause of infertility. It can be diagnosed and treated by selective salpingography and tubal catheterisation. In this paper, deficiencies of current methods of tubal assessment are discussed, compared with the relative advantages of selective salpingography and tubal catheterisation. The latter are outpatient procedures that have been shown to achieve successful fallopian tube recanalisation and pregnancies in women with infertility, without any other intervention. Another advantage of these techniques is the measurement of tubal perfusion pressure, which has a potential place in management of infertility. Finally, directions of current research into this developing subject are outlined.
Appraisal in postgraduate medical education
Authors: Fox Robert , Kane Simon
Published: February 2004

Appraisal is one important means of maximising progress under the adult learning model. With trainee and trainer working together in partnership, appraisal provides an opportunity for review of training, setting of realistic objectives and identification of training opportunities and learning methods. Appraisal is a positive, dynamic process, which provides support and praise alongside regular guidance. Appraisal cannot be expected to overcome a deficiency of learning opportunities and it must not remove the responsibility for personal development from the trainee. The initial interview is often relatively informal but a structured approach is important nevertheless. Appraisal should be viewed as an acquired skill and formal training is essential.
Vertebrate primary cilia: a sensory part of centrosomal complex in tissue cells, but a “sleeping beauty” in cultured cells?
Authors: Alieva Irina B. , Vorobjev Ivan A
Published: February 2004

Primary cilium development along with other components of the centrosome in mammalian cells was analysed ultrastructurally and by immunofluorescent staining with anti‐acetylated tubulin antibodies. We categorized two types of primary cilia, nascent cilia that are about 1 μm long located inside the cytoplasm, and true primary cilia that are several μm long and protrude from the plasma membrane.

The primary cilium is invariably associated with the older centriole of each diplosome, having appendages at the distal end and pericentriolar satellites with cytoplasmic microtubules emanating from them. Only one cilium per cell is formed normally through G0, S and G2phases. However, in some mouse embryo fibroblasts with two mature centrioles, bicilates were seen. Primary cilia were not observed in cultured cells where the mature centriole had no satellites and appendages (Chinese hamster kidney cells, line 237, some clones of l‐fibroblasts). In contrast to primary cilia, striated rootlets were found around active and non‐active centrioles with the same frequency. In proliferating cultured cells, a primary cilium can be formed several hours after mitosis, in fibroblasts 2–4 h after cell division and in PK cells only during the S‐phase. In interphase cells, formation of the primary cilium can be stimulated by the action of metabolic inhibitors and by reversed depolymerization of cytoplasmic microtubules with cold or colcemid treatments.

In mouse renal epithelial cells in situ, the centrosome was located near the cell surface and mature centrioles in 80% of the cells had primary cilium protruding into the duct lumen. After cells were explanted and subcultured, the centrosome comes closer to the nucleus and the primary cilium was depolymerized or reduced. Later primary cilia appeared in cells that form islets on the coverslip. However, the centrosome in cultured ciliated cells was always located near the cell nucleus and primary cilium never formed a characteristic distal bulb. A sequence of the developmental stages of the primary cilium is proposed and discussed. We also conclude that functioning primary cilium does not necessarily operate in culture cells, which might explain some of the contradictory data on cell ciliation in vitro reported in the literature.
Gamma irradiation of human dendritic cells influences proliferation and cytokine profile of T cells in autologous mixed lymphocyte reaction
Authors: Cao Meng‐De , Chen Zong‐De , Xing Ying
Published: February 2004

Dendritic cells (DC) are the most potent antigen‐presenting cells (APC); their ability to induce proliferation of T cells in a mixed lymphocyte reaction (MLR) assay is commonly used for the evaluation of their function. It is a general thought that gamma irradiation of APC does not influence their ability to activate T‐cell proliferation, but the data from several studies are controversial. To further determine the mechanisms involved in DC‐induced T‐cell activation in MLR assay, human DC induced from peripheral blood mononuclear cells (PBMC) were gamma‐irradiated and determine their effects on the proliferation and cytokine profiles of T cells in an autologous MLR. DC were induced from the PBMC of 11 multiple sclerosis (MS) patients with RMPI 640 medium containing recombinant human GM‐CSF (rhGM‐CSF; 800 U/ml) and recombinant human IL‐4 (rhIL‐4; 500 U/ml). DC harvested on day 7 were divided into two equal parts. One part was not irradiated (naive DC); the other was γ‐irradiated at a dose of 30 Gy. Cell surface molecules were analyzed by flow cytometry. T‐cell proliferation was determined using a beta‐scintillation counter. The levels of IL‐2, IL‐4, IL‐6 and IL‐10 in co‐culture supernatants were measured by ELISA. The results indicated that gamma irradiation reduced expression of CD86, CD80 and HLA‐DR molecules on DC, especially CD86 (P=0.0072). DC, irradiated or non‐irradiated, effectively stimulated autologous T‐cell proliferation. Compared to naive DC, irradiated DC showed a markedly lower capacity to promote T‐cell proliferation (P=0.0073), and strikingly up‐regulated secretion of IL‐4 (P=0.0145) and IL‐2 (P=0.0323) by autologous T cells. No significant differences were noted in IL‐6 and IL‐10 production between T cells co‐cultured with naive DC and irradiated DC (P>0.05). It is concluded that gamma irradiation of DC not only influences the phenotype of DC but also alters their capacity to stimulate the proliferation and the cytokine profiles of autologous T cells in a MLR.
Significance of C‐terminal sequence elements for Petunia flavanone 3β‐hydroxylase activity
Authors: Wellmann Frank , Matern Ulrich , Lukačin Richard
Published: February 2004

Flavanone 3β‐hydroxylase (FHT), a 2‐oxoglutarate‐dependent dioxygenase (2‐ODD), catalyzes the hydroxylation of (2S)‐flavanones to (2R/3R)‐dihydroflavonols in plants as a key step towards the biosynthesis of flavonols, anthocyanins and catechins. Crystallographic studies of 2‐ODDs typically revealed a jelly roll in the enzyme core, and the C‐terminus of the enzyme polypeptides was proposed to form a lid covering the active site cavity, thereby reducing the chances for oxidative or proteolytic damage and unfolding. Moreover, it has been proposed that in some cases the C‐terminus is involved in substrate selectivity of 2‐ODDs. In a systematic approach with highly active Petunia FHT, four C‐terminally truncated enzyme forms were generated by deletion of five, 11, 24 or 29 amino acids. The recombinant FHTs preserved their substrate selectivity, but the specific activity decreased gradually with the extent of truncation. Then, an enzyme chimera was constructed by domain swapping replacing the C‐terminal 52 amino acids of Petunia FHT by the equivalent region of flavonol synthase (FLS) from Citrus unshiu, an enzyme showing ambiguous FLS and FHT activity. The chimeric dioxygenase still revealed exclusively FHT activity, albeit at a moderate level only. The data predict that the selectivity of FHT is not governed by the C‐terminal sequence accounting for about 13% of the enzyme polypeptide.
Inhibition of p53 degradation by Mdm2 acetylation
Authors: Wang Xinjiang , Taplick Jan , Geva Naama , Oren Moshe
Published: February 2004

Mdm2 is a RING finger E3 ubiquitin ligase, which promotes ubiquitination and proteasomal degradation of the p53 tumor suppressor protein. Acetylation of p53 regulates p53's transcriptional activity and inhibits Mdm2‐mediated p53 ubiquitination and degradation. We now report that Mdm2 is also a target for acetylation. Mdm2 is acetylated in vitro by CREB‐binding protein (CBP) and to a lesser extent by p300, but not by p300/CPB‐associated factor. Acetylation occurs primarily within the RING finger domain of Mdm2. In vivo acetylation of Mdm2 was detected easily with CBP but not p300. Efficient in vivo acetylation required the preservation of the RING finger. An Mdm2 mutant (K466/467Q) mimicking acetylation is impaired in its ability to promote p53 ubiquitination, as well as Mdm2 autoubiquitination. Moreover, K466/467Q is defective in promoting p53 degradation in living cells. We thus suggest that acetyltransferases may modulate cellular p53 activity not only by modifying p53, but also by inactivating Mdm2.
Met144Ala mutation of the copper‐containing nitrite reductase from Alcaligenes xylosoxidans reverses the intramolecular electron transfer
Authors: Farver Ole , Eady Robert R , Sawers Gary , Prudêncio Miguel , Pecht Israel
Published: February 2004

Pulse radiolysis has been employed to investigate the intramolecular electron transfer (ET) between the type 1 (T1) and type 2 (T2) copper sites in the Met144Ala Alcaligenes xylosoxidans nitrite reductase (AxCuNiR) mutant. This mutation increases the reduction potential of the T1 copper center. Kinetic results suggest that the change in driving force has a dramatic influence on the reactivity: The T2Cu(II) is initially reduced followed by ET to T1Cu(II). The activation parameters have been determined and are compared with those of the wild‐type (WT) AxCuNiR. The reorganization energy of the T2 site in the latter enzyme was calculated to be 1.6±0.2 eV which is two‐fold larger than that of the T1 copper center in the WT protein.
Interactions between landscape changes and host communities can regulateEchinococcus multilocularistransmission
Authors: GIRAUDOUX P. , CRAIG P. S. , DELATTRE P. , BAO G. , BARTHOLOMOT B. , HARRAGA S. , QUÉRÉ J.-P. , RAOUL F. , WANG Y. , SHI D. , VUITTON D.-A.
Published: February 2004

An area close to the Qinghai-Tibet plateau region and subject to intensive deforestation contains a large focus of human alveolar echinococcosis while sporadic human cases occur in the Doubs region of eastern France. The current review analyses and compares epidemiological and ecological results obtained in both regions. Analysis of rodent species assemblages within quantified rural landscapes in central China and eastern France shows a significant association between host species for the pathogenic helminthEchinococcus multilocularis, with prevalences of human alveolar echinococcosis and with land area under shrubland or grassland. This suggests that at the regional scale landscape can affect human disease distribution through interaction with small mammal communities and their population dynamics. Lidicker's ROMPA hypothesis helps to explain this association and provides a novel explanation of how landscape changes may result in increased risk of a rodent-borne zoonotic disease.
Editorial
Authors: LIEVEN ELENA
Published: February 2004

The Editors would like to register their great sadness at the tragically early death of our colleague, Elizabeth Bates. The loss of her empirical breadth, her theoretical engagement and her sheer energy and enthusiasm for the fundamental questions that engage us all, will leave a huge gap.
State and Society in Urban China in the Wake of the 16th Party Congress
Authors: Solinger Dorothy J.
Published: February 2004

At the time of the convening of the 16th Party Congress in November 2002, the Party leadership confronted an urban society splintered by the blessings and the blows of two decades of ever-deepening marketization. This article explores the composition of urban society at that juncture, and aims to delineate its changing social structural break-down. It also investigates the correlation between Jiang Zemin's “three represents” and the various separate social groups making up the cities at the turn of the century. While arguing that in Jiang's vision the lowest stratum of society may have been intentionally excluded from his “represents,” the piece also shows the stance of the state towards several groupings and its means of dealing with each. The piece concludes with a suggestion that new top leaders Hu Jintao and Wen Jiabao appeared, as of the time they took office, to be turning a new leaf.
Privatizing Rural China: Insider Privatization, Innovative Contracts and the Performance of Township Enterprises
Authors: Li Hongbin , Rozelle Scott
Published: February 2004

This article examines the privatization of China's township enterprises. According to our survey of 670 firms in 15 randomly selected counties in Jiangsu and Zhejiang provinces, more than half of the firms owned by local government were completely privatized by 1999. The privatization process is striking for two reasons. First, local governments almost always sold firms to insiders, while in the rest of the world privatization largely involves outsiders. Secondly, unlike the predictions of some academics and policy makers, many privatized firms have experienced an increase in performance. Drawing on firm-level survey data and extensive interviews with government leaders and managers, we found that leaders devised a way to elicit information from the buyer at the time of the sale about the firm's future profitability that enabled them to execute privatization successfully. Our analysis shows that the performance of firms with new owners that paid a price for the firm that exceeded the book value of its assets is on par with the performance of private firms after privatization since they also received strong incentives.
Chinas Unlimited: Making the Imaginaries of China and Chineseness. By Gregory B. Lee. [London: RoutledgeCurzon, 2003. xi +121 pp. £65.00. ISBN 0-7007-1492-8.]
Authors: van Crevel Maghiel
Published: February 2004

Gregory Lee's latest book addresses representations of Chineseness in the 19th and 20th centuries, in the United Kingdom as well as in China and Hong Kong. Each of its four chapters singles out texts – from “high” literature, government documents, news and entertainment media – to point out the dangers of generalizing about culture, language or race. Lee observes that while the “scientific racism” of earlier days has been discredited, racist stereotypes of Chineseness abound to this day.

Chapter one begins with an explanation of the author's interest in Chinese and his career to date. This would have combined well with chapter four, subtitled “A short (hi)story of a Liverpool hybridity,” which contains scattered musings on the Liverpool Chinatown Lee knew as a child, where a lifelong fascination was sparked by a notebook left by his Chinese grandfather. Critical distance is by no means a “shibboleth” (p. 82), but Lee's lengthy justification of the autobiographical parts of his book is unnecessary. Still, the expression of his personal feeling, on the first day of each academic year, of “disappoint[ing] the eager European students as they see a white man walk into the room” (p. 83) is questionable. For all its autobiographical hues, as part of the scholarly treatise that his book aims to be, this entails the danger of unwarranted generalization.
Searching for Life's Meaning: Changes and Tensions in the Worldviews of Chinese in the 1980s. By Luo Xu. [Ann Arbor, MI: University of Michigan Press, 2002. xi +359 pp. ISBN 0-472-11239-2.]
Authors: Israel John
Published: February 2004

In analysing the youthful cohort that launched the Tiananmen protest movement, Luo Xu draws our attention to the problematic connection between young people's quest to define individual identity and the decision to commit themselves to political and social action. How, he asks, could a “self-centered ‘me generation’ . . . engage in such an enormous collective action that demanded great devotion to a common idealistic cause?” (p. ix)

Part one, “The journey” treats intellectual formulations from Bei Dao's poem I Don't Believe (1976) to Cui Jian's song I Have Nothing (1986), and summarizes public discourses by and about youth from the Democracy Wall (1978|–79) to the lesser known “Pan Xiao discussion” (1980) and “Shekou Storm” (1988). Xu's narrative implies that the tidal changes of the 13 years from the death of Mao to the Tiananmen uprising created two mini-generations. The first (late 1970s and early 1980s) was preoccupied with unresolved issues of the Cultural Revolution. The second (late 1980s) sought to redefine the relationship between the individual and the public realm.
Rethinking the 1898 Reform Period: Political and Cultural Change in Late Qing China. Edited by Rebecca E. Karl and Peter Zarrow. [Cambridge, MA and London: Harvard University Asia Center, distributed by Harvard University Press, 2002. x+273 pp. £29.95; $45.00. ISBN 0-674-00854-5.]
Authors: Thompson Roger R.
Published: February 2004

In their introduction to this excellent collection of nine essays, most of which were presented at the 1998 Annual Meeting of the Association for Asian Studies, historians Rebecca Karl and Peter Zarrow write convincingly that the “1898 period” in particular and the late Qing in general mark the moment when “Chinese experiences of modernity” (p. 10) began. Recognizing that many of the elements we associate with the May Fourth paradigm first appeared in the under-studied late-Qing period, the editors decry how the late Qing “continues to be treated in an isolated fashion and is seldom drawn into the main currents of ‘Chinese modernity,’ which are seen as more properly placed in the later May Fourth period” (p. 7). The reason to study 1898 now is that we can see and compare China's confrontations with two global capitalisms (late 19th and late 20th centuries). We don't need a “functionalist exhumation of 1898;” we need an approach that helps us understand the “local effects of globalizing trajectories” (p. 7). In sum: “[We need to] rethink 1898 not as an event per se but, more important, as a vital conjunctural historical moment, as an extended moment during and through which Chinese intellectuals and society consciously confronted and began to reformulate the Chinese historical problematic” (p. 7).