bluuebunny/crossref_metadata_2025_split huggingface dataset:
Co‐localization analysis of complex formation among membrane proteins by computerized fluorescence microscopy: application to immunofluorescence co‐patching studies
Authors: Lachmanovich E. , Shvartsman D. E. , Malka Y. , Botvin C. , Henis Y. I. , Weiss A. M. 
Published: November 2003

SummaryFluorescence imaging of two independently labelled proteins is commonly used to determine their co‐localization in cells. Antibody‐mediated crosslinking can mediate the patching of such proteins at the cell surface, and their co‐localization can serve to determine complex formation among them. However, manual analysis of such studies is both tedious and subjective. Here we present a digital co‐localization analysis that is independent of the fluorescence intensity, is highly consistent and reproducible between observers, and dramatically reduces the analysis time. The approach presented is based on a segmentation procedure that creates binary objects, and then determines whether objects belonging to two different groups (e.g. green‐ and red‐labelled) are co‐localized. Two methods are used to determine co‐localization. The ‘overlap’ analysis defines two objects as co‐localized if the centre of mass of one falls within the area of the other. The ‘nearest‐neighbour distance’ analysis considers two objects as co‐localized if their centres are within a threshold distance determined by the imaging modality. To test the significance of the results, the analysis of the actual images is tested against randomized images generated by a method that creates images with uncorrelated distributions of objects from the two groups. The applicability of the algorithms presented to study protein interactions in live cells is demonstrated by co‐patching studies on influenza haemagglutinin mutants that do or do not associate into mutual oligomers at the cell surface via binding to AP‐2 adaptor complexes. The approach presented is potentially applicable to studies of co‐localization by other methods (e.g. electron microscopy), and the nearest‐neighbour distance method can also be adapted to study phenomena of correlated placement.
Monitoring cell movements and volume changes with pulse‐mode scanning ion conductance microscopy
Authors: Happel P. , Hoffmann G. , Mann S. A. , Dietzel I. D. 
Published: November 2003

SummaryHere we describe the use of pulse‐mode scanning ion conductance microscopy (SICM) to observe volume changes and cell membrane movements during the locomotion of cultured cells in the range of minutes to several hours. The microscope is based on the pulse‐mode SICM previously developed for stable imaging of single cells in culture. Our instrument uses current pulses to control the distance between cell surface and electrode tip as well as a back‐step mode to prevent contact of tip and membrane during lateral movements of the probe. We performed repeated scans of cell surfaces using feedback‐controlled piezoactors to position the electrode. Using patch‐clamp‐type electrode tips the height of cells could reproducibly be measured with a standard deviation of 50 nm. To quantify and separate changes in cell position and volume occurring between consecutive scans, a program was written to subtract images and calculate volume changes. Examples of repeated scans show that membrane movements in the range of 30 min to a few hours can be quantitatively monitored with a lateral resolution of 500 nm using difference images and that faster movements in the range of minutes can be recorded at defined cell sections using the line scan mode. Difference images indicate that volume changes can affect cell surfaces inhomogeneously, emphazising the role of the cytoskeleton in the stabilization of cell shape.
Basal cell carcinomas of covered and unusual sites of the body
Authors: Betti Roberto , Bruscagin Cinzia , Inselvini Elena , Crosti Carlo 
Published: November 2003

Abstract Background The concept of unusual site for a basal cell carcinoma (BCC) has to be defined because there is much confusion about it. Basal cell carcinomas of covered areas of the body are sometimes considered as unusual because of their location. In this study an attempt is made to define what is an unusual site for a BCC according to experience and the problems encountered are briefly discussed. Methods A total of 1050 histologically‐confirmed primary BCCs were studied. Age, sex and site of presentation were considered. Results Eighteen cases were recorded, localized on hand, axilla, areola or nipple, buttock, perineal, and genital regions. Conclusions Unusual location has been arbitrarily defined in the past according to the incidental percentage, to a mathematical index of density, to the literature reports. All these criteria suffer more or less from some discrepancy, but in our opinion all these should be considered for a definition. Our results, moreover, suggest that only the above described sites should be considered.
Granuloma faciale: successful treatment of nine cases with a combination of cryotherapy and intralesional corticosteroid injection
Authors: Dowlati Bijan , Firooz Alireza , Dowlati Yahya 
Published: November 2003

Abstract Background Granuloma faciale is a rare disorder characterized by asymptomatic papules, nodules, and plaques on the face. Although the exact pathogenesis of this disease is unclear, it is considered a variant of leukocytoclastic vasculitis confined to the skin. Several medical and surgical methods have been used to treat it with variable results. Case reports We report nine cases of granuloma faciale treated with a combination of cryotherapy with liquid nitrogen and intralesional injection of corticosteroids. Results The lesions cleared completely in all of the patients without any side‐effects. No recurrences have been observed. Conclusions Cryotherapy with liquid nitrogen, followed by intralesional injection of corticosteroids, is a safe and effective method to treat granuloma faciale.
Effect of the vehicle on the amount of stratum corneum removed by tape stripping
Authors: Jacobi Ute , Meykadeh Nuschin , Sterry Wolfram , Lademann Jürgen 
Published: November 2003

Summary Background: The penetration of topically applied substances into the stratum corneum can non‐invasively be studied using the tape stripping procedure. This method was applied to investigate in vivo the penetration of a fragrance, vanillin, applied in ethanol and a w/o emulsion. Methods: Twenty tape strips were removed from each skin area treated with vanillin in ethanol or w/o emulsion, respectively. The concentration of vanillin was determined for each tape strip. In addition, the pseudo‐absorption of the corneocytes was determined to calculate the SC profile. Results: The vanillin concentration was correlated both with the tape number and with the stratum corneum profile. Depending on whether the tape number or the profile of the stratum corneum were correlated with the vanillin concentration, different distributions within the stratum corneum were obtained. Different amounts of stratum corneum were removed with 20 tape strips dependent on the vehicle applied previously. The application of the w/o emulsion led to the removal of nearly the half the amount of corneocytes stripped from the ethanol‐treated area. Conclusions: The results obtained underline the general necessity to correlate the amount of stratum corneum with the amount of substance in penetration studies.
Sperm Supply and Egg fertilization in the Ostrich (Struthio camelus)
Authors: Malecki IA , Martin GB 
Published: November 2003

ContentsWe used egg break‐out and spermatozoa trapped in the perivitelline layer of eggs to test the hypothesis that sperm supply and egg fertilization rate are high in the ostrich. Egg fertilization status was determined at break‐out by the appearance of the germinal disc (GD) and then the perivitelline layer overlying the GD region was collected to count sperm (SpermOPVL) under fluorescence following staining with 4′,6′‐diamidino‐2‐phenyindole (DAPI). The study was carried out on commercial ostrich farms over two laying seasons. In the first year, 229 eggs from nine randomly chosen ostrich pens comprising pairs, trios (two females, one male) and larger groups were collected for 1 week of every month of laying. Eggs contained 253 ± 18 SpermOPVL/mm2 of the GD (mean ± SEM; range 0–1330). Egg fertilization rate averaged 89.4 ± 3.4% and varied from 78.6 to 98.2% between pens. Month had no effect on sperm supply or egg fertilization status. Eggs from paired birds (sex ratio 1 : 1) had less sperm in the GD than the eggs from pens with a higher sex ratio. In the second year, 150 eggs from seven pens, each containing only one male and either one, two or three females, were studied for 2 weeks at the beginning (winter), middle (spring) and end (summer) of laying. Eggs contained 364 ± 45 SpermOPVL/mm2 of the GD (range 0–2880). Season had no effect on sperm supply or egg fertilization. The number of SpermOPVL varied between pens, assumed to be due to variation between individual males. The number of SpermOPVL increased as the sex ratio increased only when very high‐ranking males were excluded from the analysis. Egg fertilization rate was 94.4 ± 3.1% but varied from 64.0 to 100% between pens. Egg fertilization was not affected by season or sex ratio. Low fertilization rates were observed in two pens and appeared related to the lack of synchrony between timing of laying and sperm production in the first, and lack of mating in the second pen. We conclude that ostrich flocks generally have high rates of egg fertilization and any infertility is associated with lack of sperm supply.
Successful treatment of in‐transit melanoma metastases using topical 2‐4 dinitrochlorobenzene
Authors: Von Nida Jamie , Quirk Chris 
Published: November 2003

SUMMARYA 78‐year‐old woman presented with multiple histologically proven in‐transit melanoma metastases involving the lower half of the left leg. Initial therapy with liquid nitrogen cryotherapy had some short‐lived success but was not tolerated by the patient. As further lesions began to develop, daily topical application of 2% 2‐4 dinitrochlorobenzene to the lesions was commenced. During the first 2 years of therapy a partial response was achieved, with treated lesions regressing while new lesions developed. Eventually a long‐term remission of almost 2 years with no clinical evidence of cutaneous melanoma deposits was achieved. This treatment did not prevent metastatic lymph node and ultimately fatal liver involvement. Topical immunotherapy can be a useful adjunct in the treatment of cutaneous melanoma metastases, particularly in those patients who are unable to tolerate other destructive modalities of therapy.
Assessing Assessing! An Examination of the Impact of the New RANZCP by-Laws for Training
Authors: Malhi Gin , McKay Diana 
Published: November 2003

Objectives: With the Royal Australian and New Zealand College of Psychiatrists’ new by-laws for training coming into effect in late 2003 and early 2004 for New Zealand and Australian trainees, respectively, the authors set out to determine the impact of the various proposed changes upon assessment. Methods: The relevant medical databases and websites were explored and the various colleges and examination boards in Australia and New Zealand, the UK and Canada were consulted. Results: The Fellowship examination in Australia and New Zealand is due to undergo significant changes in terms of its structure and methods of assessment. The changes borrow components from contemporary international systems and aim to shift the emphasis of training and examination to earlier in the postgraduate years, with trainees ultimately having greater flexibility in terms of deciding when to sit for these. As yet, there is little specific information for candidates as regards the exact format and content of some components of the examinations and it is unclear whether the new system has been evaluated sufficiently. Conclusions: Many of the proposed changes bear similarities to training structures and examination systems in Canada and the UK. It is suggested that the ‘mistakes’ of others should be learned from, and that a considered and deliberate approach is taken to the introduction of new examinations, appraising their merit and acknowledging the difficulties that change and uncertainty create.
The Domestic Sources of Differences in Labour Market Policies
Authors: Hamann Kerstin , Kelly John 
Published: November 2003

AbstractIn this article we analyse what drives both the specific choices and the relative amount of change in labour market policies in two countries, Britain and Spain, in the 1990s. Although we find more substantial and radical labour market reform in Britain, Spain experienced greater union inclusion in policy‐making. We examine the roles of economic institutions and the ideology of the governing party and find that neither accounts satisfactorily for the labour market changes in our cases. Domestic political variables, especially electoral factors and the power and autonomy of government, give more insight into the similarities and differences between the two countries.
International Political Economy and Industrial Relations
Authors: Haworth Nigel , Hughes Stephen 
Published: November 2003

AbstractThe field of international political economy offers valuable insights into global economic integration, trade liberalization, global governance and the nature and activities of international organizations and regimes. Despite their impact on industrial relations, industrial relations theory has not wholeheartedly engaged with these phenomena or this sister field. This paper argues that the field of international political economy offers much to industrial relations, particularly in terms of understanding the nature and impact of internationalization and emerging global governance. The potential in a closer relationship between the two fields is illustrated by the example of the international labour standards regime and its principal organization, the International Labour Organisation.
Definition of the microvascular pattern of the normal human adult mammary gland
Authors: Naccarato Antonio G. , Viacava Paolo , Bocci Guido , Fanelli Giovanni , Aretini Paolo , Lonobile Antonino , Montruccoli Giancarlo , Bevilacqua Generoso 
Published: November 2003

AbstractThe present report provides a detailed description of microvascularization in the normal human mammary gland, and defines two novel morphometric parameters to be used as a reference when angiogenesis in breast carcinoma is evaluated. Microvascularization was analysed by histology, immunohistochemistry and computer‐assisted analysis in a set of breast tissue samples taken from nine women, during the pre‐ovulatory phase of the menstrual cycle. The two parameters designed for image analysis were: vascular density (VD): [microvessel number/(microvessel area + residual stromal area)] × 10 000; and vascular area ratio (VAR): [microvessel area/(microvessel area + residual stromal area)]. In the lobules VD (mean value ± SE 2.48 ± 0.14) and VAR (0.33 ± 0.02) showed little variability and correlated significantly (P < 0.05). The areas occupied by microvessels, stroma and acini remained constant in all lobules (21.53 ± 1.87%, 42.65 ± 1.35% and 35.14 ± 1.57%, respectively). Microvascularization of the lobules was of a sinusoidal type, with large S‐shaped capillaries. In the ducts VD (2.95 ± 0.16) and VAR (0.29 ± 0.03) showed little variability but did not correlate significantly. Microvascularization of the ducts was of a classic type, with capillaries normal in size and shape. The expression of oestrogen (ER) and progesterone (PR) receptors was analysed by immunohistochemistry and compared with the morphometric results. ER expression levels were in the range 20–25% (24.3 ± 2.1) and 14–18% (15.4 ± 1.5) in lobules and ducts, respectively. PR expression levels were in the range 10–13% (11.1 ± 1.6) and 14–17% (15.2 ± 1.4), respectively. No correlation was found between ER/PR expression and vascularization parameters.
Origin and development of the pronephros in the chick embryo
Authors: Hiruma Tamiko , Nakamura Hiroaki 
Published: November 2003

AbstractThe process by which the pronephros develops was morphologically examined in chick embryos from Hamburger–Hamilton stage (ST) 8+ to ST34. The intermediate mesoderm, from which the pronephros arises, was first seen as a faint ridge of undifferentiated mesoderm between the segmental plate and lateral plate at ST8+. It formed a cell cord at the level of the 6th to the presumptive 13th somites at ST9 to ST10. This cell cord then separated into dorsal and ventral parts, the former becoming the nephric duct and the latter the tubules by ST14. The primordia of the external glomeruli (PEGs) appeared at ST15 through some epithelial cells protruding in the nephrostome (the opening of the nephric tubule into the body cavity). PEGs formed gradually in the caudal direction until ST18, while the pronephric tubules and PEGs in cranial locations disappeared. At this stage, only a few PEGs remained at the level of the 13th and 14th somites and these developed from ST23 to ST29 to become ultrastructurally similar to the glomeruli of the functional kidney. From these observations in the avian pronephros, we infer that the pronephric duct and tubules both form from a cell cord in the intermediate mesoderm and at the same time, but later develop differently.
Glucagon‐like peptide 1 modulates calcium responses to glutamate and membrane depolarization in hippocampal neurons
Authors: Gilman Charles P. , Perry TracyAnn , Furukawa Katsotoshi , Grieg Nigel H. , Egan Josephine M. , Mattson Mark P. 
Published: November 2003

AbstractGlucagon‐like peptide 1 (GLP‐1) activates receptors coupled to cAMP production and calcium influx in pancreatic cells, resulting in enhanced glucose sensitivity and insulin secretion. Despite evidence that the GLP‐1 receptor is present and active in neurons, little is known of the roles of GLP‐1 in neuronal physiology. As GLP‐1 modulates calcium homeostasis in pancreatic beta cells, and because calcium plays important roles in neuronal plasticity and neurodegenerative processes, we examined the effects of GLP‐1 on calcium regulation in cultured rat hippocampal neurons. When neurons were pre‐treated with GLP‐1, calcium responses to glutamate and membrane depolarization were attenuated. Whole‐cell patch clamp analyses showed that glutamate‐induced currents and currents through voltage‐dependent calcium channels were significantly decreased in neurons pre‐treated with GLP‐1. Pre‐treatment of neurons with GLP‐1 significantly decreased their vulnerability to death induced by glutamate. Acute application of GLP‐1 resulted in a transient elevation of intracellular calcium levels, consistent with the established effects of GLP‐1 on cAMP production and activation of cAMP response element‐binding protein. Collectively, our findings suggest that, by modulating calcium responses to glutamate and membrane depolarization, GLP‐1 may play important roles in regulating neuronal plasticity and cell survival.
Prolonged exposure of microglia to lipopolysaccharide modifies the intracellular signaling pathways and selectively promotes prostaglandin E2 synthesis
Authors: Ajmone‐Cat M. A. , Nicolini A. , Minghetti L. 
Published: November 2003

AbstractDuring inflammatory or degenerative processes microglial cells are likely to be exposed to activating agents that persist in brain parenchyma for prolonged periods. As our knowledge on microglial activation is largely based on in vitro studies in which microglial cultures are activated by a single administration of pro‐inflammatory stimuli, we investigated the effects of repeated endotoxin (LPS) challenges on microglial functional state. Primary rat microglial cultures were subjected to one, two or three consecutive LPS‐stimulation and the production of tumor necrosis factor‐α (TNF‐α), nitric oxide (NO), prostaglandin E2 (PGE2) and 15‐deoxy‐Δ12,14‐PGJ2 (15d‐PGJ2) measured. The ability of microglial cells to produce NO, TNF‐α and 15d‐PGJ2 upon the first LPS challenge rapidly declined after the second and the third stimulations, whereas PGE2 synthesis remained constantly elevated. Accordingly, the expression of inducible NO synthase decreased whereas cyclooxygenase‐2 and microsomal PGE synthase remained up‐regulated. The signaling pathways evoked by single or multiple LPS‐stimulation were also profoundly different, when considering the activation of the transcription factors nuclear factor‐kappa B and CREB, and of the p38 MAPK. Our observations suggest that prolonged exposure to LPS, and likely other activating agents, induces in microglia a functional state clearly distinct from that triggered by acute stimulation. The progressive down‐regulation of pro‐inflammatory molecules and the sustained release of PGE2 could have important implications for the resolution of brain inflammation.
Identification of a region from the human cholinergic gene locus that targets expression of the vesicular acetylcholine transporter to a subset of neurons in the medial habenular nucleus in transgenic mice
Authors: Schütz Burkhard , Damadzic Ruslan , Weihe Eberhard , Eiden Lee E. 
Published: November 2003

AbstractWe use a transgenic mouse model system to elucidate the regulatory regions within the human cholinergic gene locus responsible for vesicular acetylcholine transporter gene expression in vivo. In this report we characterized two transgenes for their ability to confer cholinergic‐specific expression of the encoded vesicular acetylcholine transporter. An 11.2 kb transgene (named hV11.2) that spanned from about 5 kb upstream of the start of vesicular acetylcholine transporter translation down to the first choline acetyltransferase coding exon gave expression in the somatomotor neurons and a subpopulation of cholinergic neurons in the medial habenular nucleus. The second transgene (named hV6.7), a 5‐prime truncated version of hV11.2 that was devoid of 4.5 kb of gene‐regulatory sequences completely lacked vesicular acetylcholine transporter expression in vivo. Our data indicate that vesicular acetylcholine transporter expression in somatomotor neurons and in the medial habenular nucleus is uniquely specified within the cholinergic gene locus, and separable from cholinergic expression elsewhere. The identification of these two subdivisions of the cholinergic nervous system suggests that other cholinergic neurons in the CNS and PNS are similarly regulated by additional discrete domains within the cholinergic gene locus.
Isolation and characterization of microsatellite DNA markers from the Lanyu scops owl (Otus elegans botelensis)
Authors: Hsu Yu‐Cheng , Severinghaus Lucia L. , Lin Yao‐Sung , Li Shou‐Hsien 
Published: November 2003

AbstractFrom a genomic library enriched for GATA/CTAT and GAAA/CTTT repeats, 12 polymorphic microsatellite markers were developed for the Lanyu scops owl (Otus elegans botelensis). Polymorphism of these loci was evaluated in a sample of 58 adult individuals of unknown relationship. The allele numbers of each locus were from five to 25 and the observed heterozygosity of each locus ranges from 0.707 to 0.914. When using these loci in parentage assignment, the probability of false parent exclusion is greater than 0.999. All loci conformed to Hardy–Weinberg expectations.
Development of microsatellite markers for the European white elm (Ulmus laevis Pall.) and cross‐species amplification within the genus Ulmus
Authors: Whiteley R. E. , Black‐Samuelsson S. , Clapham D. 
Published: November 2003

Abstract Ulmus laevis Pall. is a broad‐leaved deciduous tree with a central and eastern European distribution. We describe the development of six polymorphic microsatellite markers for this species. These markers were also tested for utility in U. americana, U. glabra, U. minor and U. pumila. One additional marker gave ambiguous results in U. laevis but amplified clearly in three other species. In U. laevis, the number of alleles observed per locus ranged from two to nine. Five loci showed polymorphism in at least one of the nontarget species tested.