bluuebunny/crossref_metadata_2025 huggingface dataset:
88 PREGNANCY RATES FOR IN VITRO AND IN VIVO PRODUCED OVINE EMBRYOS VITRIFIED USING THE MINIMUM VOLUME COOLING CRYOTOP METHOD
Authors: Kelly J. , Kleemann D. , Kuwayama M. , Walker S.
Published: December 2004

Previously we reported that, using the minimum volume cooling (MVC) cryotop vitrification method, in vitro-produced ovine and bovine embryo survival after thawing was similiar to that of fresh embryos (Kelly et al. 2004 Reprod. Fert. Dev. 16, 172). While survival of vitrified embryos after thawing can be indicative of embryo viability, this assessment does not always correlate with embryo survival after transfer. This study assesses the effect of vitrification using the MVC cryotop method on the survival after transfer of in vitro- and in vivo-produced ovine embryos. Fresh or vitrified Day 6 ovine embryos (expanded blastocysts, blastocysts, compact morulae) were used in this study. Ovine cumulus–oocyte complexes were obtained and matured, fertilized (Day 0), and cultured in vitro (Walker et al. 1996 Biol. Reprod. 55, 703–708). In vitro embryos for vitrification were produced and vitrified (Kelly et al. 2004 Reprod. Fert. Dev. 16, 172) 10 days prior to the day of transfer. In vivo embryos were recovered from donor Merino ewes and vitrified 7 days prior to the day of transfer while fresh in vivo embryos were collected and transferred on the same day. Semen used for both in vivo and in vitro embryo production was from the same sire. On the day of transfer, vitrified embryos were thawed directly into 1.25 M sucrose solution, followed by stepwise dilution of the cryoprotectants. Embryos were transferred as singles into synchronized recipient ewes on a randomized basis. Fetal number was detected at Day 50. Variables were assessed using the CATMOD procedure in SAS. Pregnancy rate for in vivo-derived embryos was higher (P < 0.01) than for in vitro-derived embryos. Embryo treatment (fresh vs. vitrified) did not significantly affect pregnancy rate. Pregnancy rate for ewes detected (by vasectomized rams) in estrus within 48 h of progesterone pessary removal was higher (P < 0.05) than for both the 48–68 h and unmarked groups. The latter two groups did not differ significantly. None of the first-order interactions were significant (P > 0.05). This study demonstrates that ovine embryos (in vitro and in vivo) can be vitrified, thawed, and transferred without compromising embryo viability. However, the differences in pregnancy rate between the recipient groups warrant further investigation. The MVC cryotop method is a vitrification technique that can be adapted to routine field use.

Table 1.
Pregnancy rate of fresh and vitrified in vivo and in vitro ovine embryos after embryo transfer
Force measurements in E-cadherin–mediated cell doublets reveal rapid adhesion strengthened by actin cytoskeleton remodeling through Rac and Cdc42
Authors: Chu Yeh-Shiu , Thomas William A. , Eder Olivier , Pincet Frederic , Perez Eric , Thiery Jean Paul , Dufour Sylvie
Published: December 2004

We have used a modified, dual pipette assay to quantify the strength of cadherin-dependent cell–cell adhesion. The force required to separate E-cadherin–expressing paired cells in suspension was measured as an index of intercellular adhesion. Separation force depended on the homophilic interaction of functional cadherins at the cell surface, increasing with the duration of contact and with cadherin levels. Severing the link between cadherin and the actin cytoskeleton or disrupting actin polymerization did not affect initiation of cadherin-mediated adhesion, but prevented it from developing and becoming stronger over time. Rac and Cdc42, the Rho-like small GTPases, were activated when E-cadherin–expressing cells formed aggregates in suspension. Overproduction of the dominant negative form of Rac or Cdc42 permitted initial E-cadherin–based adhesion but affected its later development; the dominant active forms prevented cell adhesion outright. Our findings highlight the crucial roles played by Rac, Cdc42, and actin cytoskeleton dynamics in the development and regulation of strong cell adhesion, defined in terms of mechanical forces.
Quantitative mass spectrometry reveals a role for the GTPase Rho1p in actin organization on the peroxisome membrane
Authors: Marelli Marcello , Smith Jennifer J. , Jung Sunhee , Yi Eugene , Nesvizhskii Alexey I. , Christmas Rowan H. , Saleem Ramsey A. , Tam Yuen Yi C. , Fagarasanu Andrei , Goodlett David R. , Aebersold Ruedi , Rachubinski Richard A. , Aitchison John D.
Published: December 2004

We have combined classical subcellular fractionation with large-scale quantitative mass spectrometry to identify proteins that enrich specifically with peroxisomes of Saccharomyces cerevisiae. In two complementary experiments, isotope-coded affinity tags and tandem mass spectrometry were used to quantify the relative enrichment of proteins during the purification of peroxisomes. Mathematical modeling of the data from 306 quantified proteins led to a prioritized list of 70 candidates whose enrichment scores indicated a high likelihood of them being peroxisomal. Among these proteins, eight novel peroxisome-associated proteins were identified. The top novel peroxisomal candidate was the small GTPase Rho1p. Although Rho1p has been shown to be tethered to membranes of the secretory pathway, we show that it is specifically recruited to peroxisomes upon their induction in a process dependent on its interaction with the peroxisome membrane protein Pex25p. Rho1p regulates the assembly state of actin on the peroxisome membrane, thereby controlling peroxisome membrane dynamics and biogenesis.
The integrin effector PINCH regulates JNK activity and epithelial migration in concert with Ras suppressor 1
Authors: Kadrmas Julie L. , Smith Mark A. , Clark Kathleen A. , Pronovost Stephen M. , Muster Nemone , Yates John R. , Beckerle Mary C.
Published: December 2004

Cell adhesion and migration are dynamic processes requiring the coordinated action of multiple signaling pathways, but the mechanisms underlying signal integration have remained elusive. Drosophila embryonic dorsal closure (DC) requires both integrin function and c-Jun amino-terminal kinase (JNK) signaling for opposed epithelial sheets to migrate, meet, and suture. Here, we show that PINCH, a protein required for integrin-dependent cell adhesion and actin–membrane anchorage, is present at the leading edge of these migrating epithelia and is required for DC. By analysis of native protein complexes, we identify RSU-1, a regulator of Ras signaling in mammalian cells, as a novel PINCH binding partner that contributes to PINCH stability. Mutation of the gene encoding RSU-1 results in wing blistering in Drosophila, demonstrating its role in integrin-dependent cell adhesion. Genetic interaction analyses reveal that both PINCH and RSU-1 antagonize JNK signaling during DC. Our results suggest that PINCH and RSU-1 contribute to the integration of JNK and integrin functions during Drosophila development.
Developmentally regulated alterations in Polycomb repressive complex 1 proteins on the inactive X chromosome
Authors: Plath Kathrin , Talbot Dale , Hamer Karien M. , Otte Arie P. , Yang Thomas P. , Jaenisch Rudolf , Panning Barbara
Published: December 2004

Polycomb group (PcG) proteins belonging to the polycomb (Pc) repressive complexes 1 and 2 (PRC1 and PRC2) maintain homeotic gene silencing. In Drosophila, PRC2 methylates histone H3 on lysine 27, and this epigenetic mark facilitates recruitment of PRC1. Mouse PRC2 (mPRC2) has been implicated in X inactivation, as mPRC2 proteins transiently accumulate on the inactive X chromosome (Xi) at the onset of X inactivation to methylate histone H3 lysine 27 (H3-K27). In this study, we demonstrate that mPRC1 proteins localize to the Xi, and that different mPRC1 proteins accumulate on the Xi during initiation and maintenance of X inactivation in embryonic cells. The Xi accumulation of mPRC1 proteins requires Xist RNA and is not solely regulated by the presence of H3-K27 methylation, as not all cells that exhibit this epigenetic mark on the Xi show Xi enrichment of mPRC1 proteins. Our results implicate mPRC1 in X inactivation and suggest that the regulated assembly of PcG protein complexes on the Xi contributes to this multistep process.
PCB assembly
Authors: Golding David
Published: December 2004

Discusses the manufacturing technology associated with the assembly of
printed circuit boards [PCBs] covering photolithography, etching,
manual assembly of PCBs and the soldering methods involved. Describes how all
the stages of PCB assembly can be automated and also looks at how the boards
are cleaned and tested. Concludes that many factors need to be taken into
account when deciding which method of manufacturing printed circuits would be
best suited to a company’s requirements.
Gendered Violence: Castration and Blinding as Punishment for Treason in Normandy and Anglo‐Norman England
Authors: Van Eickels Klaus
Published: December 2004

The use of castration as a punishment for treason and other forms of misdemeanour was a specific trait of the Norman realms of medieval Europe. In the post‐Carolingian kingdoms of France, Germany and Italy, it was rarely practised and only known as a punishment for sexual crimes. In Scandinavia, Normandy, Anglo‐Norman England and Norman Sicily, however, blinding and castration were regarded as an appropriate equivalent of the death penalty. The particular emphasis on masculinity implied in the Norman construction of noble honour, rendered the Norman warrior's body particularly vulnerable. Since his testicles were regarded as the prerequisite of his social existence, they became a legitimate point of attack whenever the ruler felt betrayed and decided to use force against his enemies. This gendered violence constituted a constantly renewed frame of reference, which defined political power as male and reinforced the notion that authority required a fully functional masculine body.
Mannose binding lectin and C3 act as recognition molecules for infectious agents in the vagina
Authors: Pellis V , De Seta F , Crovella S , Bossi F , Bulla R , Guaschino S , Radillo O , Garred P , Tedesco F
Published: December 2004

In our study we examined the early complement components in patients with bacterial vaginosis (BV), vulvovaginal candidiasis (VVC) and in healthy controls. The levels of C1q, mannose-binding lectin (MBL) and C3 were measured by ELISA in the cervicovaginal lavage (CVL) from gynaecological patients and controls. No significant differences were observed in the levels of these proteins in the three study groups. Immunofluorescence analysis of the clue cells and Candida hyphae from BV and VVC patients for surface-bound complement components showed the presence of C3, while C1q was undetectable. MBL was revealed on clue cells but not on Candida. Binding of MBL to Candida, grown or cytocentrifuged from the CVL of VVC patients, was found to be pH dependent and occurred between pH 4·5 and pH 5·5. In conclusion, we demonstrated that MBL and C3 present in the vaginal cavity act as recognition molecules for infectious agents that colonize the cervicovaginal mucosa. Our finding that MBL, but not C1q, binds to bacteria and fungi in vagina suggests that the lectin and classical pathways of complement activation may play a different role in immune defence in the female genital tract.
Immunosuppression: towards a logical approach in liver transplantation
Authors: Perry I , Neuberger J
Published: December 2004

Over the last two decades there has been a significant increase in the number and types of immunosuppressive agents that have been available to clinicians. The protocols for immunosuppression used in liver transplantation have been derived historically from those in renal transplantation. During the last decade there has been a shift in the use of immunosuppression, with the introduction of interleukin (IL)-2 receptor antagonists in place of anti-lymphocyte preparations, substitution of tacrolimus for cyclosporin and mycophenolate for azathioprine. The use of corticosteroids has been reduced. For a variety of reasons, these changes have not always been made on the basis of properly randomized studies. The place of newer agents, such as sirolimus and leflunomide derivatives and of the microbiological agents, is unclear. In this review, we outline briefly the mechanism of action of drugs and suggest possible approaches to the management of the liver allograft recipient, suggesting how treatment could be adjusted according to the indication for transplantation as well as the individual's comorbidities.
CD26 (dipeptidyl-peptidase IV)-dependent recruitment of T cells in a rat asthma model
Authors: Kruschinski C , Skripuletz T , Bedoui S , Tschernig T , Pabst R , Nassenstein C , Braun A , Von Hörsten S
Published: December 2004

CD26 truncates several chemokines as well as neuropeptides and influences immune responses via modulation of cell adhesion and T cell activation, suggesting an involvement of CD26 in asthmatic and airway inflammation. Therefore, Fischer 344 (F344), Brown Norway (BN) and Lewis (LEW) rat strains, which differ in their CD26-like enzymatic activity, were compared using an asthma model. Additionally, two CD26-deficient mutant F344 rat substrains were included and compared to the wild-type F344 substrain. Immunization was performed twice with ovalbumin (OVA), and 2 weeks later the rats were challenged with OVA intratracheally Flow cytometry (FACS) analysis of different leucocyte subsets as well as enzyme-linked immunosorbent assay (ELISA) for IgE levels in the blood and bronchoalveolar lavage (BAL) were performed 24 h after challenge. LEW rats with the lowest CD26 activity among the rat strains investigated here displayed significantly reduced CD4+ T cell numbers in the BAL compared to wild-type F344 and BN rats. Moreover, in asthma, the ratio of CD26+ to CD26– T cell receptor (TCR)-positive cells increased significantly in F344 and LEW but not BN rats. Most intriguingly, in both CD26-deficient F344 rat substrains the number of CD4+ T lymphocytes was markedly reduced compared to wild-type F344. The decrease in T cell recruitment observed in the CD26-deficient rats was associated with significantly reduced OVA-specific IgE-titres. This is the first report to show a remarkably reduced T cell recruitment in rat strains that either lack or exhibit reduced CD26-like enzymatic activity, suggesting a role for CD26 in the pathogenesis of asthma via T cell-dependent processes such as antibody production.
Relevance ofex vivoblood lymphocyte assay forin vivolymphocyte function
Authors: Brattig N W , Timmann C , Abraha R S , Lepping B , Müller-Myhsok B , Horstmann R D
Published: December 2004

Determinations of in vitro proliferative and secretory activities of peripheral blood cells are used widely for research in clinical immunology but, to our knowledge, have not been evaluated as to their power to reflect in vivo activities quantitatively. Here, we addressed this question by quantitatively correlating the in vitro secretion of interleukin (IL)-5 by peripheral blood cells to the in vivo activity of IL-5 as reflected by peripheral-blood eosinophil counts. Studying 458 humans exposed to transmission of the nematode Onchocerca volvulus, IL-5 was measured in the supernatants of 0·02-ml whole-blood cells cultured in the presence of O. volvulus extract or mitogen. O. volvulus-reactive IL-5 secretion was correlated significantly to blood eosinophilia in a quantitative manner explaining 15·1% (95% CI 8·3–19·9%) of the variability of eosinophil counts. Interestingly, correlations were obtained only if parasite counts were included in the calculation using multiple regression analysis. The results show that in vitro assays of minute amounts of blood lymphocytes may quantitatively reflect activities of the entire lymphocyte population in vivo.
Role of p38 MAPK and NF-kB for chemokine release in coculture of human eosinophils and bronchial epithelial cells
Authors: Wong C K , Wang C B , Ip W K , Tian Y P , Lam C W K
Published: December 2004

Eosinophils are principal effector cells of inflammation in allergic asthma, characterized by their accumulation and infiltration at inflammatory sites mediated by the chemokine eotaxin and their interaction with adhesion molecules expressed on bronchial epithelial cells. We investigated the modulation of nuclear factor-κB (NF-κB) and the mitogen-activated protein kinase (MAPK) pathway on the in vitro release of chemokines including regulated upon activation normal T cell expressed and secreted (RANTES), monokine induced by interferon-γ (MIG), monocyte chemoattractant protein-1 (MCP-1), interleukin (IL)-8, and interferon-inducible protein-10 (IP-10) upon the interaction of human bronchial epithelial BEAS-2B cells and eosinophils. Gene expression of chemokines was evaluated by RT-PCR and the induction amount of chemokines quantified by cytometric bead array. NF-κB and p38 MAPK activities were assessed by electrophoretic mobility shift assay and Western blot, respectively. The interaction of eosinophils and BEAS-2B cells was found to up-regulate the gene expression of the chemokines IL-8, MCP-1, MIG, RANTES and IP-10 expression in BEAS-2B cells, and to significantly elevate the release of the aforementioned chemokines except RANTES in a coculture of BEAS-2B cells and eosinophils. IκB-α phosphorylation inhibitor, BAY 11–7082, and p38 MAPK inhibitor, SB 203580 could decrease the release of IL-8, IP-10 and MCP-1 in the coculture. Together, the above results show that the induction of the release of chemokines in a coculture of epithelial cells and eosinophils are regulated by p38 MAPK and NF-κB activities of BEAS-2B cells, at least partly, through intercellular contact. Our findings therefore shed light on the future development of more effective agents for allergic and inflammatory diseases.
Effects of systemic infections with Tobacco rattle virus on agronomic and quality traits of a range of potato cultivars
Authors: Dale M. F. B. , Robinson D. J. , Todd D.
Published: December 2004

Infection of several commercially important potato cultivars with Tobacco rattle virus (TRV) can result in systemic infection with M‐type virus, which persists through generations of vegetative propagation, although spraing symptoms in the tubers rarely develop. In field trials of nine such cultivars it was shown that TRV infection can delay plant emergence and retard subsequent growth, reduce tuber yield and mean tuber weight, increase tuber numbers and the proportion of small tubers, increase the incidence of growth cracks and misshapen tubers, diminish dry matter content, and exacerbate after‐cooking blackening. Each cultivar was affected differently, showing different combinations and degrees of these effects. Virus concentration in leaf extracts from individual plants was also highly variable. These, and other potato cultivars similarly susceptible to TRV, are grown on large areas of UK and European potato land. The implications of growing such cultivars for the seed, ware and processing sectors of the potato industry are discussed.
Real‐time PCR assay for quantification of Tilletia caries contamination of UK wheat seed
Authors: McNeil M. , Roberts A.M.I. , Cockerell V. , Mulholland V.
Published: December 2004

A quantitative real‐time PCR assay using TaqMan chemistry has been developed to quantify the level of Tilletia spp. contamination in wheat‐seed lots. In the UK wheat seed is predominantly contaminated with Tilletia caries (syn. Tilletia tritici), and the probability of detecting other Tilletia spp. is negligible. DNA standards, prepared from T. caries spores, were calibrated using a set of 26 seed samples, with T. caries contamination levels ranging from 0 to 1000 spores per seed. The linear calibration model obtained by the regression of log10 (number of spores per seed + 1) on mean log10 DNA (µg) produced a coefficient of determination (R2) of 0·904. The calibration model was tested using 226 seed samples; of these, 91% fell within the 95% confidence intervals. Of the 21 samples that were outside the limits, 16 were overpredictions and five underpredictions. The five underpredictions were all from seed samples where contamination was less than one spore per seed. The model predicts that samples with 44 pg of DNA will be below one spore per seed with 95% probability. Of the 226 test samples compared with this threshold, 99 contained less than 44 pg DNA, and these were found to have less than one spore per seed by microscopic assay. This real‐time assay allows an increase in test throughput and provides the sensitivity required for an advisory threshold of one spore per seed.
Populus genotypes differ in infection by, and systemic spread of, Poplar mosaic virus
Authors: Smith C. M. , Campbell M. M.
Published: December 2004

The spatial and temporal dynamics of Poplar mosaic virus (PopMV) infection were investigated using four different genotypes of Populus. Four Populus clones were vegetatively propagated under sterile conditions and used as hosts for PopMV infection in controlled growth‐room experiments. The presence of PopMV RNA was assessed using reverse transcriptase PCR. The presence of infectious PopMV particles was assessed by the ability of extracts from Populus leaves to generate PopMV symptoms on the propagation host, Nicotiana megalosiphon. Neither PopMV RNA nor infectious particles accumulated to detectable levels in one of the four clones (CT6009), suggesting that this clone had preformed resistance to PopMV. PopMV RNA and infectious particles were restricted to the inoculated leaf in two of the clones (AP37 and NM6), suggesting that these clones restricted either the rate of virus accumulation or the spread of PopMV itself. In contrast, PopMV systemically infected one clone (52‐226), indicating that this host was either fully susceptible, or had delayed resistance.
A perspective on the measurement of time in plant disease epidemiology
Authors: Lovell D. J. , Powers S. J. , Welham S. J. , Parker S. R.
Published: December 2004

The growth and development of plant pathogens and their hosts generally respond strongly to the temperature of their environment. However, most studies of plant pathology record pathogen/host measurements against physical time (e.g. hours or days) rather than thermal time (e.g. degree‐days or degree‐hours). This confounds the comparison of epidemiological measurements across experiments and limits the value of the scientific literature.
Expression of carbohydrate xenoantigens on porcine peripheral nerve
Authors: Magnusson Stefan , Strokan Valeri , Svensson Lola , Månsson Jan‐Eric , Rydberg Lennart , Breimer Michael E.
Published: December 2004

Abstract:  Background: 
The use of thin easily revascularized cutaneuos nerve autografts, which has been the gold standard, or the alternative use of nerve allografts or artificial grafts for nerve reconstructing have all their pros and cons. Nerve xenotransplantation may offer a potential alternative. In a potential pig to human nerve xenograft transplantation set‐up several porcine antigen barriers have to be considered such as carbohydrate antigens system like the blood group A/O, the Galα1‐3Gal (αGal) and the Hanganutziu‐Deicher (HD) antigens. The swine leukocyte protein antigens system may also have to bee considered. The knowledge of the antigen expression on pig peripheral nerves is today limited.

The present study describes the distribution of glycolipid based carbohydrate xenoantigens in ischiadicus nerve from blood group A and O pigs.

Methods:  Glycolipid fractions were separated on thin layer chromatography plates and immunostained with human AB sera, biotinylated Griffonia simplicifolia isolectin B4, monoclonal antibodies reacting with the HD antigen and with blood group A antigens based on different core saccharide structures. In addition, the subcellular distribution of αGal and HD antigens were studied by light‐ and electron‐microscopical immunohistochemistry. The total amount of neutral glycolipids was 15 mg/g tissue for both blood group A and O nerves with mono‐glycosylceramides as the dominating component.

Results and conclusions:  The total amount of acidic glycolipids (gangliosides and sulpholipids) was 9 mg/g tissue for both the blood group O and A nerves with sulphatides as the dominating components. Analyses of the glycolipid fractions showed strong expression of both the αGal and the HD antigens in nerves from both blood group A and O pigs. In addition, small amounts of blood group A antigens were expressed in nerves from blood group A pigs. Staining of neutral glycolipids from blood group A pigs using monoclonal antibodies reacting with A antigen having different core structures suggested that the A epitope expressed on pig ischiadicus nerves is based on the type 1 core chain structure. Light and electron microscopical studies on the αGal and HD‐antigen distribution revealed that the neural cells were αGal antigen negative. Endothelial cells of blood vessels, and lymphatic and perineural cells expressed αGal antigen. Both endothelial cells and myelinized axons revealed positively labelled for the HD antigen.
Early weaning of piglets fails to exclude porcine lymphotropic herpesvirus
Authors: Mueller Nicolas J. , Kuwaki Kenji , Knosalla Christoph , Dor Frank J. M. F. , Gollackner Bernd , Wilkinson Robert A. , Arn Scott , Sachs David H. , Cooper David K. C. , Fishman Jay A.
Published: December 2004

Abstract:  Background:  Xenotransplantation using pigs as source species carries a risk for the activation of latent herpesviruses from the porcine donor and potential transmission to the recipient. In pig‐to‐baboon xenotransplantation, activation of porcine cytomegalovirus (PCMV) has been associated with xenograft injury and an increased incidence of consumptive coagulopathy and graft loss. Activation of porcine lymphotropic herpesvirus (PLHV)‐1 was not observed in pig‐to‐baboon solid organ xenotransplantation, but was associated with a syndrome of post‐transplantation lymphoproliferative disorder (PTLD) after allogeneic stem cell transplantation in pigs.

Material and Methods:  Early weaning of piglets was used to try to reduce the viral burden of xenograft donors. This consisted of separating the piglets of a litter from the sow within the first 2 weeks after birth and raising them in isolation from the remaining herd.

Results:  We have previously demonstrated that PCMV could be excluded from source animals by early weaning of piglets. However, early weaning failed to exclude PLHV‐1 from source pigs.

Conclusions:  This disparity between PCMV and PLHV‐1 reflects differing pathogenesis of infection of these herpesviruses. New approaches will be needed to exclude PLHV‐1 from pig colonies.
Phaiodotoxin, a novel structural class of insect‐toxin isolated from the venom of the Mexican scorpion Anuroctonus phaiodactylus
Authors: Valdez‐Cruz Norma A. , Batista Cesar V. F. , Zamudio Fernando Z. , Bosmans Frank , Tytgat Jan , Possani Lourival D.
Published: December 2004

A peptide called phaiodotoxin was isolated from the venom of the scorpion Anuroctonus phaiodactylus. It is lethal to crickets, but non toxic to mice at the doses assayed. It has 72 amino acid residues, with a molecular mass of 7971 atomic mass units. Its covalent structure was determined by Edman degradation and mass spectrometry; it contains four disulfide‐bridges, of which one of the pairs is formed between cysteine‐7 and cysteine‐8 (positions Cys63–Cys71). The other three pairs are formed between Cys13–Cys38, Cys23–Cys50 and Cys27–Cys52. Comparative sequence analysis shows that phaiodotoxin belongs to the long‐chain subfamily of scorpion peptides. Several genes coding for this peptide and similar ones were cloned by PCR, using cDNA prepared from the RNA of venomous glands of this scorpion. Electrophysiological assays conducted with this toxin in several mammalian cell lines (TE671, COS7, rat GH3 and cerebellum granular cells), showed no effect on Na+ currents. However, it shifts the voltage dependence of activation and inactivation of insect Na+ channels (para/tipE) to more negative and positive potentials, respectively. Therefore, the ‘window’ current is increased by 225%, which is thought to be the cause of its toxicity toward insects. Phaiodotoxin is the first toxic peptide ever purified from a scorpion of the family Iuridae.
Classification of ATP‐dependent proteases Lon and comparison of the active sites of their proteolytic domains
Authors: Rotanova Tatyana V. , Melnikov Edward E. , Khalatova Anna G. , Makhovskaya Oksana V. , Botos Istvan , Wlodawer Alexander , Gustchina Alla
Published: December 2004

ATP‐dependent Lon proteases belong to the superfamily of AAA+ proteins. Until recently, the identity of the residues involved in their proteolytic active sites was not elucidated. However, the putative catalytic Ser–Lys dyad was recently suggested through sequence comparison of more than 100 Lon proteases from various sources. The presence of the catalytic dyad was experimentally confirmed by site‐directed mutagenesis of the Escherichia coli Lon protease and by determination of the crystal structure of its proteolytic domain. Furthermore, this extensive sequence analysis allowed the definition of two subfamilies of Lon proteases, LonA and LonB, based on the consensus sequences in the active sites of their proteolytic domains. These differences strictly associate with the specific characteristics of their AAA+ modules, as well as with the presence or absence of an N‐terminal domain.
Importance of evacuatory disturbance in evaluation of faecal incontinence after third degree obstetric tear
Authors: Shatari T. , Hayes J. , Pretlove S. , Toosz‐Hobson P. , Radley S. , Keighley M. R. B.
Published: December 2004

Objective  To correlate anorectal function including rectal evacuation with anorectal physiology and endoanal ultrasound in women with third degree obstetric anal sphincter injury repaired at the time of delivery.

Patients and methods  Forty‐four women with repaired third degree tears underwent anorectal physiology, anal ultrasonography and clinical assessment using the St. Marks incontinence score (0–24). Evacuatory disturbance was assessed by questionnaire.

Results  There was a significant correlation between disturbed evacuation and incontinence symptoms (P =0.030). There was also a significant correlation between disturbed evacuation and internal anal sphincter (IAS) injury (P = 0.026), but there was no correlation with external anal sphincter (EAS) injury. There was a correlation between disturbed evacuation and low resting anal pressure (P = 0.013). Although IAS defects were associated with low anal pressure, only the correlation with Maximum Squeeze Pressure reached statistical significance (P = 0.018).

Conclusion  Women with evacuatory disturbance after repaired third degree tears have a greater level of incontinence than those whose emptying is normal. This association is related to internal sphincter injury and reduced anal sphincter pressures.
Hospital volume and outcome of rectal cancer surgery in Denmark 1994–99
Authors: Harling H. , Bülow S. , Møller L. N. , Jørgensen T. ,   the Danish Colorectal Cancer Group
Published: December 2004

Objective  The association between hospital volumen and outcome of major cancer surgery is being debated at present. We analysed the outcome of rectal cancer surgery in Denmark during the period 1994–99.

Methods  All patients with a first‐time rectal cancer were registered in a national database during the 5‐year period. In this observational cohort study, the influence of hospital case volume on resectional procedure, complications, 30‐day mortality and 5‐year mortality was analysed.

Results  The register comprised 5021 patients. Surgery was performed in 27 hospitals with < 15 operations per year, 15 hospitals with 15–30 operations per year and 11 hospitals with > 30 operations per year. In a multivariate model, the risk of permanent colostomy was significantly increased in the group of low‐volume hospitals. On the contrary, volume did not influence the risk of anastomotic leakage, 30‐day mortality and 5‐year mortality. However, a large variation in 5‐year mortality was observed particularly within the low‐volume group of hospitals.

Conclusions  In this study, only risk of having a permanent colostomy during surgery for rectal cancer was significantly related to hospital case volume. When individual hospitals were analysed, a large variation in 5‐year mortality was observed within the low‐volume group of hospitals.
Usefulness of total cholesterol/HDL‐cholesterol ratio in the management of diabetic dyslipidaemia
Authors: Gimeno‐Orna J. A. , Faure‐Nogueras E. , Sancho‐Serrano M. A.
Published: December 2004

Aims  Our aim was to evaluate the usefulness of the total cholesterol/HDL‐cholesterol ratio (TC/HDL) in predicting the cardiovascular risk of Type 2 diabetic patients.

Methods  Prospective cohort study with inclusion of 418 Type 2 diabetic individuals with follow‐up until the appearance of a cardiovascular event. The predictive power of updated mean lipid values during follow‐up was analysed by means of Cox proportional hazard models. An estimate was made of the relative risk (RR) conferred by high levels of TC/HDL stratified by LDL‐cholesterol levels.

Results  Sixty‐six cardiovascular events occurred during an average follow‐up of 4.7 years (sd 1.5). The main lipid predictor of vascular events was mean TC/HDL ratio [hazard ratio (HR) = 1.46; 95% confidence interval (CI) 1.25, 1.7; P < 0.0001]. In the multivariate model with simultaneous inclusion of mean TC/HDL and mean LDL‐cholesterol, both were significant predictors of cardiovascular disease [HR (1 unit) = 1.37; 95% CI 1.16, 1.62; P = 0.0003 and HR (1 mmol/l) = 1.5; 95% CI 1.04, 2.18; P = 0.03, respectively]. The LDL adjusted RR for cardiovascular events due to high TC/HDL ratio, with 4.5 cut‐off point, was 2.5 (95% CI 1.4, 4.3; P = 0.0007). For the stratum of subjects with average LDL cholesterol < 3.5 mmol/l, RR was 1.2 (95% CI 0.5, 2.8; P = NS) and for the stratum of average LDL cholesterol > 3.5 mmol/l, RR was 4 (95% CI 1.8, 9; P = 0.00013), with heterogeneity among strata (P = 0.044).

Conclusions  It could be useful to include the TC/HDL ratio in treatment guides for diabetic dyslipidaemia, given their high predictive value and strong interaction with LDL cholesterol.
Insulino‐mimetic and anti‐diabetic effects of vanadium compounds
Authors: Srivastava A. K. , Mehdi M. Z.
Published: December 2004

Compounds of the trace element vanadium exert various insulin‐like effects in in vitro and in vivo systems. These include their ability to improve glucose homeostasis and insulin resistance in animal models of Type 1 and Type 2 diabetes mellitus. In addition to animal studies, several reports have documented improvements in liver and muscle insulin sensitivity in a limited number of patients with Type 2 diabetes. These effects are, however, not as dramatic as those observed in animal experiments, probably because lower doses of vanadium were used and the duration of therapy was short in human studies as compared with animal work. The ability of these compounds to stimulate glucose uptake, glycogen and lipid synthesis in muscle, adipose and hepatic tissues and to inhibit gluconeogenesis, and the activities of the gluconeogenic enzymes: phosphoenol pyruvate carboxykinase and glucose‐6‐phosphatase in the liver and kidney as well as lipolysis in fat cells contributes as potential mechanisms to their anti‐diabetic insulin‐like effects. At the cellular level, vanadium activates several key elements of the insulin signal transduction pathway, such as the tyrosine phosphorylation of insulin receptor substrate‐1, and extracellular signal‐regulated kinase 1 and 2, phosphatidylinositol 3‐kinase and protein kinase B activation. These pathways are believed to mediate the metabolic actions of insulin. Because protein tyrosine phosphatases (PTPases) are considered to be negative regulators of the insulin‐signalling pathway, it is suggested that vanadium can enhance insulin signalling and action by virtue of its capacity to inhibit PTPase activity and increase tyrosine phosphorylation of substrate proteins. There are some concerns about the potential toxicity of available inorganic vanadium salts at higher doses and during long‐term therapy. Therefore, new organo‐vanadium compounds with higher potency and less toxicity need to be evaluated for their efficacy as potential treatment of human diabetes.
The TGF‐β1 gene codon 10 polymorphism contributes to the genetic predisposition to nephropathy in Type 1 diabetes
Authors: Patel A. , Scott W. R. , Lympany P. A. , Rippin J. D. , Gill G. V. ,   the Warren 3/UK GoKind Study Group, Barnett A. H. , Bain S. C.
Published: December 2004

Aims  We hypothesize that transforming growth factor‐beta (TGF‐β), a multifunctional growth factor which plays a key role in the development of tissue fibrosis, may be involved in the pathophysiology of diabetic nephropathy. Our aim was to examine three polymorphisms within the TGF‐β1 gene, in codons 10, 25 and 263, for association with nephropathy in Type 1 diabetes.

Methods  We conducted a large case–control study using cases with Type 1 diabetes and clinical nephropathy. Controls were Type 1 diabetic subjects who have been injecting insulin for at least 50 years and have extremely low risk of nephropathy. Genotyping was by polymerase chain reaction with sequence‐specific primers.

Results  There was a significant difference in the frequency of the TGF‐β1 codon 10 genotypes in the diabetic nephropathy group (n = 420) when compared with the controls (n = 410, P = 0.007). There were no significant differences when the frequencies of the TGF‐β1 codons 25 and 263 genotypes in the diabetic nephropathy group were compared with the control group.

Conclusions  In our study the TGF‐β1 codon 10 polymorphism is associated with nephropathy in Type 1 diabetes and variation in this gene may contribute to the genetic predisposition to this complication in Type 1 diabetes.
Conceptualising State Capacity: Comparing Kazakhstan and Kyrgyzstan
Authors: Cummings Sally N. , Nørgaard Ole
Published: December 2004

Strengthening the state is central to the post-communist reform agenda. Here, state capacity combines organisational, material and social resources and is conceptualised along four dimensions: ideational, political, technical and implementational. This conceptualisation is applied to a comparative, survey-based analysis in 2002 of 125 medium-ranking officials in two post-communist Central Asian countries, Kazakhstan and Kyrgyzstan. The findings reveal that although Kazakhstan's controlled economic reform programme and natural resources have placed it in a stronger position to develop its state capacity, important ideational, political and implementational problems pose long-term obstacles for reform. In turn, Kyrgyzstan's early liberalisation in the absence of economic and social resources may be serving to undermine its state capacity.
Political Philosophy and Empowering Citizens
Authors: de-Shalit Avner
Published: December 2004

This paper defends the idea of empowering citizens by means of teaching them political philosophy. First, I explain and define empowerment as an experience leading to the development of critical and philosophical capabilities. Several challenges to using philosophy to empower citizens are then discussed and rejected. This group of challenges is called the ‘divorce theory’, because, according to them, philosophy and politics should be distinguished, as if divorced from each other, so that they can live happily side by side, but not together. Finally, empowerment is normatively defended and distinguished from paternalism, and examine the relationships between empowerment through political philosophy and deliberative democracy.
Early onset severe pre‐eclampsia: expectant management at a secondary hospital in close association with a tertiary institution
Authors: Oettle Charl , Hall David , Roux Adele , Grové Debbie
Published: December 2004

Objectives  Early onset severe pre‐eclampsia is ideally managed in a tertiary setting. We investigated the possibility of safe management at secondary level, in close co‐operation with the tertiary centre.

Design  Prospective case series over 39 months.

Setting  Secondary referral centre.

Population  All women (n= 131) between 24 and 34 weeks of gestation with severe pre‐eclampsia, where both mother and fetus were otherwise stable.

Methods  After admission, frequent intensive but non‐invasive monitoring of mother and fetus was performed. Women were delivered on achieving 34 weeks, or if fetal distress or major maternal complications developed. Transfer to the tertiary centre was individualised.

Main outcome measures  Prolongation of gestation, maternal complications, perinatal outcome and number of tertiary referrals.

Results  Most women [n= 116 (88.5%)] were managed entirely at the secondary hospital. Major maternal complications occurred in 44 (33.6%) cases with placental abruption (22.9%) the most common. One maternal death occurred and two women required intensive care admission. A mean of 11.6 days was gained before delivery with the mean delivery gestation being 31.8 weeks. The most frequent reason for delivery was fetal distress (55.2%). There were four intrauterine deaths. The perinatal mortality rate (≥1000 g) was 44.4/1000, and the early neonatal mortality rate (≥500 g) was 30.5/1000.

Conclusions  The maternal and perinatal outcomes are comparable to those achieved by other tertiary units. This model of expectant management of early onset, severe pre‐eclampsia is encouraging but requires close co‐operation between secondary and tertiary institutions. Referrals to the tertiary centre were optimised, reducing their workload and costs, and patients were managed closer to their communities.
Bladder mucosal autografts for repair of vesicovaginal fistula
Authors: Vyas Nachiket , Nandi Priya Ranjan , Mahmood Mufti , Tandon Vipul , Dwivedi Udai Shankar , Singh Pratap Bahadur
Published: December 2004

Twenty‐two women with primary and secondary (five patients) vesicovaginal fistula attending a tertiary level urological unit in India were treated by repair of the fistula using bladder mucosal autografts. The fistula was approached transabdominally or via a combined abdominal and vaginal approach (for those involving the trigone). After closure of the vaginal layer, bladder mucosa was harvested from the dome of the bladder and laid over the fistula with sutures at each corner to fix it in place. Patients were catheterised for 12–14 days. At follow up after 3 to 12 months, 20 out of 22 patients were continent, with no other symptoms. The two failures had undergone two previous repairs each. This series is the first from India, and demonstrates the efficacy of bladder mucosal autografts for managing large fistulae, those where a previous repair has failed and fistulae adjacent to the ureteric orifice without the need for uretero‐neocystostomy.
Effects of long term administration of KUR‐1246, a selective β2‐adrenoceptor agonist, on pregnant sheep and their fetuses
Authors: Murata Satoshi , Matsuda Tadashi , Kiguchi Sumiyoshi , Kobayashi Mamoru , Cho Kazutoshi , Okuyama Kazuhiko
Published: December 2004

Objective  To evaluate the safety of KUR‐1246 as a tocolytic agent, we examined the effects of its long term infusion on respiratory and cardiovascular systems and general metabolism in pregnant sheep and their fetuses.

Design  Animal experiment with chronically instrumented ewes and their fetuses.

Setting  Center for animal experiments, Hokkaido University School of Medicine, Japan.

Sample  Eight Suffolk ewes at 117 to 120 days of gestation.

Methods  At 120–124 days of gestation, ewes (n= 4) were infused intravenously for 24 hours with KUR‐1246 at 0.03 μg/kg/minute, a dose that completely inhibits oxytocin‐induced uterine contractions in pregnant sheep. The controls received saline instead (n= 4). Statistical comparisons were carried out by repeated‐measures ANOVA followed by Dunnett's test.

Main outcome measures  Maternal and fetal values of heart rate, blood pressure, plasma electrolytes, glucose, insulin and non‐esterified fatty acid levels, and blood gases and lactate level.

Results  The maternal plasma levels of KUR‐1246 increased and reached a plateau at 15 hours or later from the start of the infusion, whereas the fetal levels of it were below the lower limit of quantification (0.1 ng/mL) throughout the experiment. Significant differences over time between the ewes that had received with KUR‐1246 and the controls were found for the following parameters: maternal heart rate, blood lactate, plasma glucose, and plasma insulin levels, and fetal plasma glucose and plasma insulin levels (P < 0.05). In the KUR‐1246 treated ewes, significant changes from the pre‐infusion value were detected in maternal blood lactate and fetal plasma glucose levels within 6 hours from the start of the infusion (P < 0.05). No significant differences were observed in other parameters in either ewes or fetuses.

Conclusion  The physiologic changes induced by a 24‐hour infusion of KUR‐1246 were transient and considered to be within the compensatory capacity in both pregnant ewes and their fetuses, suggesting that KUR‐1246 is a potentially safe tocolytic agent for use by long term infusion.
Gedrite–garnet–sillimanite‐bearing granulites from Amessmessa area, south In Ouzzal, Hoggar, Algeria
Authors: OUZEGANE K. , DJEMAI S. , GUIRAUD M.
Published: December 2004

Some granulites from the Amessmessa area (south In Ouzzal unit, Hoggar) contain the peak assemblage gedrite+garnet+sillimanite+quartz that was used to estimate the P–T conditions of metamorphism. The rocks developed symplectites and corona textures by the breakdown of the primary paragenesis to orthopyroxene, cordierite and spinel. The successive parageneses formed in separate microdomains according to a clockwise P–T path. Geothermometry, geobarometry and phase diagram calculations indicate that the textures formed by decompression and cooling from 7–9 kbar and 850–900°C to 3.5–4.5 kbar and 700–800°C. This P–T evolution is consistent with low to medium aH2O, between 0.4 and 0.7, and is similar to the metamorphic conditions deduced in Al–Mg granulites from the north of In Ouzzal.