bluuebunny/crossref_metadata_2025 huggingface dataset:
Parameterization and Evaluation of a Spatiotemporal Model of the Potato Late Blight Pathosystem
Authors: Skelsey P. , Kessel G. J. T. , Rossing W. A. H. , van der Werf W.
Published: February 2009

A spatiotemporal, integrodifference equation model of the potato late blight pathosystem is described. Formerly, the model was used in a theoretical context to analyze and predict epidemic dynamics in spatially heterogeneous mixtures of host genotypes. The model has now been modified to reflect a research interest in interactions between genotype, environment, landscape, and management. New parameter values describing host–pathogen interactions were determined and new environment–pathogen relationships included. A new analytical equation describing lesion expansion and associated necrosis has also been developed. These changes prompted a need to assess the quality of model predictions. Cultivar–isolate-specific interactions were characterized in the model using three quantitative components of resistance: infection efficiency, lesion growth rate, and sporulation intensity. These were measured on detached potato leaflets in the laboratory. Results of a sensitivity analysis illuminate the effect of different quantitative components of resistance and initial conditions on the shape of disease progress curves. Using the resistance components, the epidemic process of lesion expansion was separated from the epidemic process of lesion propagation providing two reference curves for diagnosing observed epidemics. The spatial component of the model was evaluated graphically in order to determine if realistic rates of focal expansion for potato late blight are produced. In accordance with theory, the radius of a predicted focus increased linearly with time and a constant focal velocity was reached that compared well with published experimental data. Validation data for the temporal model came from 20 late blight epidemics observed in field trials conducted in the Netherlands in 2002 and 2004. The field data and model were compared visually using disease progress curves, and numerically through a comparison of predicted and observed t5 and t50 points (time in days until 5 and 50% disease severity is reached, respectively) and relative areas under the disease progress curve values. Temporal model predictions were in close agreement with observational data and the ability of the model to translate measured resistance components, weather data, and initial conditions into realistic disease progress curves without the need for calibration confirms its utility as a tool in the analysis and diagnosis of epidemics.
Discrimination Between Mild and Severe Citrus tristeza virus Isolates with a Rapid and Highly Specific Real-Time Reverse Transcription-Polymerase Chain Reaction Method Using TaqMan LNA Probes
Authors: Ruiz-Ruiz S. , Moreno P. , Guerri J. , Ambrós S.
Published: February 2009

Severe isolates of Citrus tristeza virus (CTV) inducing seedling yellows (SY) and/or stem pitting (SP) in grapefruit or sweet orange are a major threat for the citrus industry worldwide. Identification of these CTV variants was achieved by quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) using a general primer set and three TaqMan locked nucleic acids (LNA) probes targeting sequences characteristic of severe, mild (non-SY, non-SP), and T36-like isolates. Successful amplification was achieved from fresh or silica-desiccated CTV-infected samples and all isolates but one reacted with one or more probes. Standard curves using RNA transcripts homologous to the three probes allowed a reproducible quantitative assay, with a wide dynamic range of detection starting with 102 copies. RT-PCR assays with homologous and heterologous transcript RNA mixes demonstrated that each probe reacted only with its cognate sequence which was detected even at ratios below 2.5%. Analysis of 56 pathogenically distinct CTV isolates from 20 countries showed that mild isolates reacted only with the mild probe, whereas severe SP and SY isolates reacted with the severe-SP or the T36-like probes, respectively, and often with a second probe. This procedure can be useful to identify and control potentially dangerous CTV isolates in areas affected only by mild isolates.
Wnt signaling pathways meet Rho GTPases
Authors: Schlessinger Karni , Hall Alan , Tolwinski Nicholas
Published: February 2009

Wnt ligands and their receptors orchestrate many essential cellular and physiological processes. During development they control differentiation, proliferation, migration, and patterning, while in the adult, they regulate tissue homeostasis, primarily through their effects on stem cell proliferation and differentiation. Underpinning these diverse biological activities is a complex set of intracellular signaling pathways that are still poorly understood. Rho GTPases have emerged as key mediators of Wnt signals, most notably in the noncanonical pathways that involve polarized cell shape changes and migrations, but also more recently in the canonical pathway leading to β-catenin-dependent transcription. It appears that Rho GTPases integrate Wnt-induced signals spatially and temporally to promote morphological and transcriptional changes affecting cell behavior.
EpidemAAITO: Features of food allergy in Italian adults attending allergy clinics: a multi‐centre study
Authors: Asero R. , Antonicelli L. , Arena A. , Bommarito L. , Caruso B. , Crivellaro M. , De Carli M. , Della Torre E. , Della Torre F. , Heffler E. , Lodi Rizzini F. , Longo R. , Manzotti G. , Marcotulli M. , Melchiorre A. , Minale P. , Morandi P. , Moreni B. , Moschella A. , Murzilli F. , Nebiolo F. , Poppa M. , Randazzo S. , Rossi G. , Senna G. E.
Published: February 2009

Background
Studies of the prevalence of different types of food allergy in adults are lacking.

Objective
To define the prevalence of IgE‐mediated food allergies in Italian adults attending allergy clinics and to assess possible differences associated with geographical position and/or dietary habits.

Methods
Seventeen allergy outpatient clinics scattered throughout Italy participated to a multi‐centre study in 2007. The number of atopic subjects and of food allergic patients along with clinical features were recorded by pre‐defined criteria. Patients with unequivocal history of food allergy confirmed by positive skin prick test were included as cases.

Results
Twenty five thousand six hundred and one subjects were screened; 12 739 (50%) were atopic, and 1079 (8,5%) had IgE‐mediated food allergy. Sixty four percent of patients were females. Overall, the most frequent food allergy was the pollen–food allergy syndrome (55%), which was associated with oral allergy syndrome in 95% of cases and whose frequency decreased southbound. Forty‐five percent of patients had a type 1 food allergy, in most cases (72%) caused by fruits and vegetables, and generally associated with a history of systemic symptoms. Type 1 food allergies represented 96% of food allergies in the South. Lipid transfer protein (LTP) accounted for 60% of sensitizations and caused most primary food allergies in all areas.

Conclusion
Plant‐derived foods cause most food allergies in Italian adults. The pollen–food allergy syndrome is the most frequent type of food allergy followed by allergy to LTP whose frequency increases southbound. The pattern of allergy to certain foods is clearly influenced by specific geographic features such as pollen exposure and dietary habits.
The companions: regulatory T cells and gene therapy
Authors: Eghtesad Saman , Morel Penelope A. , Clemens Paula R.
Published: February 2009

Undesired immunological responses to products of therapeutic gene replacement have been obstacles to successful gene therapy. Understanding such responses of the host immune system to achieve immunological tolerance to a transferred gene product is therefore crucial. In this article, we review relevant studies of immunological responses to gene replacement therapy, the role of immunological tolerance mediated by regulatory T cells in down‐regulating the unwanted immune responses, and the interrelationship of the two topics.
The relationship between human effector and memory T cells measured by ex vivo and cultured ELISPOT following recent and distal priming
Authors: Todryk Stephen M. , Pathan Ansar A. , Keating Sheila , Porter David W. , Berthoud Tamara , Thompson Fiona , Klenerman Paul , Hill Adrian V. S.
Published: February 2009

Maintenance of T‐cell responses is an essential feature in protection from many infectious diseases that must be harnessed in vaccination. The relationship between effector T‐cell responses and more durable and highly proliferative T‐cell memory, particularly in humans, is not well understood. In this study, effector T‐cell responses were measured by overnight ex vivo interferon‐γ (IFN‐γ) enzyme‐linked immunosorbent spot‐forming cell assay (ELISPOT), whereas memory T cells were measured by 10‐day culture followed by IFN‐γ ELISPOT (cultured ELISPOT). We observed a significant correlation between IFN‐γ responses to CD4‐stimulatory, but not to CD8‐stimulatory, recall antigens measured by these assays, suggesting a divergence in regulation. In vaccine trial participants who received a prime‐boost vaccination regimen comprising malaria antigens delivered by poxviruses, there was a correlation between ex vivo and cultured responses on day 7, but not 3 months post‐vaccination, with the ratio of cultured : ex vivo response increasing over time. To compare responses revealed by cultured ELISPOT in more detail, tetramers comprising viral recall antigens were used to ascribe effector‐memory and central‐memory T‐cell phenotypes through CCR7 and CD62L costaining. For CD8+ responses the effector phenotype decreased during the initial culture period and memory populations remained high within the resulting 20‐fold to 50‐fold increased IFN‐γ‐secreting or tetramer+ population. This was less marked for CD4+ responses, which had higher starting memory phenotype. Depletion of these central‐memory T‐cell populations generally ablated responses in cultured ELISPOT and reduced ex vivo responses. This study highlights differences between CD4+ and CD8+ effector and memory T cells, and the more complex phenotype of CD4+ T cells.
Comparison of pain thresholds and pain tolerance levels between Middle Easterners and Swedes and between genders
Authors: DAWSON A. , LIST T.
Published: February 2009

Summary  Objective: This study evaluates the presence of culture and gender differences in pain thresholds and pain tolerance levels between Middle Easterners and Swedes. Methods: Sixty‐four healthy individuals, 32 Middle Easterners (16 men and 16 women, mean age: 24·6 ± 3·4 years) and 32 Swedes (16 men and 16 women, mean age: 24 ± 3·5 years) participated in the study. Three experimental pain tests were conducted in each participant. Pain thresholds and pain tolerance levels were measured using an algometer (mechanical stimulus), the PainMatcher® (electric stimulus) and cold pressor test (thermal stimulus). Results: While no significant differences in pain thresholds were observed between Middle Easterners and Swedes in algometer and cold pressor tests, differences in pain tolerance levels were significant (P < 0·01 for both tests). All between‐culture differences in pain perception, pain threshold and pain tolerance level were non‐significant when measured with the PainMatcher. Significant between‐gender differences were observed only in pain threshold with the PainMatcher (P < 0·05) and in pain tolerance level with the algometer (P < 0·01) and the PainMatcher (P <0·001). Conclusion: This study found significant differences in two out of three pain tolerance level tests – but not pain threshold tests – between the Middle Eastern and Swedish cultures and between genders. These differences were more pronounced between Middle Eastern and Swedish men than between Middle Eastern and Swedish women. Gender differences were more pronounced within the Swedish than the Middle Eastern culture. These findings indicate that culture and gender influence pain experience.
Effect of age on the enteric nervous system of the human colon
Authors: Bernard C. E. , Gibbons S. J. , Gomez‐pinilla P. J. , Lurken M. S. , Schmalz P. F. , Roeder J. L. , Linden D. , Cima R. R. , Dozois E. J. , Larson D. W. , Camilleri M. , Zinsmeister A. R. , Pozo M. J. , Hicks G. A. , Farrugia G.
Published: February 2009

Abstract  The effect of age on the anatomy and function of the human colon is incompletely understood. The prevalence of disorders in adults such as constipation increase with age but it is unclear if this is due to confounding factors or age‐related structural defects. The aim of this study was to determine number and subtypes of enteric neurons and neuronal volumes in the human colon of different ages. Normal colon (descending and sigmoid) from 16 patients (nine male) was studied; ages 33–99. Antibodies to HuC/D, choline acetyltransferase (ChAT), neuronal nitric oxide synthase (nNOS), and protein gene product 9.5 were used. Effect of age was determined by testing for linear trends using regression analysis. In the myenteric plexus, number of Hu‐positive neurons declined with age (slope = −1.3 neurons/mm/10 years, P = 0.03). The number of ChAT‐positive neurons also declined with age (slope = −1.1 neurons/mm/10 years of age, P = 0.02). The number of nNOS‐positive neurons did not decline with age. As a result, the ratio of nNOS to Hu increased (slope = 0.03 per 10 years of age, P = 0.01). In the submucosal plexus, the number of neurons did not decline with age (slope = −0.3 neurons/mm/10 years, P = 0.09). Volume of nerve fibres in the circular muscle and volume of neuronal structures in the myenteric plexus did not change with age. In conclusion, the number of neurons in the human colon declines with age with sparing of nNOS‐positive neurons. This change was not accompanied by changes in total volume of neuronal structures suggesting compensatory changes in the remaining neurons.
Rocket Immunoelectrophoresis
Authors: Weeke Bent
Published: February 2009

Synonyms: —antigen electrophoresis in antibody‐containing agarose gel (84)

—electroimmuno assay (85).
14. Quantitation of Plasma Proteins in Cerebrospinal Fluid
Authors: BOCK ELISABETH
Published: February 2009

Summary. Fifteen plasma proteins were determined both in unconcentrated cerebrospinal fluid (CSF) and in serum from 22 patients suffering from various psychiatric and neurological disorders, by means of rocket immunoelectrophoresis. The procedure is described and the values are reported. The concentration of the individual proteins in the CSF is correlated to the concentration in serum and to the total protein content of the CSF. It is concluded that the CSF protein pattern can only be evaluated if the CSF total protein is known and if the plasma protein pattern in blood is investigated by means of, e.g., an electrophoresis in agarose gel.
Complexes in Human Plasma between α1‐Antitrypsin and IgA, and α1‐Antitrypsin and Fibrinogen
Authors: LAURELL C.‐B. , THULIN E.
Published: February 2009

Crossed immunoelectrophoretic analyses revealed that plasma contains a complex between α1‐antitrypsin1 and IgA. Normally the complex constitutes 1–2% of these proteins. The amount varies mainly with the IgA and to some extent with the α1‐at content of the plasma. The complex is sensitive to thiol reagents and is reduced in vivo by penicillamine. The size of the complex and the amount of reduction products are compatible with the assumption of 1 IgA monomer or dimer per α1‐at molecule. An α1‐at complex with fibrinogen is also a regular plasma constituent. This complex is not susceptible to thiol reagents.
Immunochemical Studies of Lipoproteins in Cholestasis
Authors: EKMAN R. , JOHANSSON B. G. , PETERSSON B.‐G.
Published: February 2009

Lipoproteins in normal and cholestatic human plasma were studied using crossed immuno‐electrophoresis and electroimmunoassay. Crossed immuno‐electrophoresis against anti‐α‐lipoprotein revealed a complex pattern of α‐lipoproteins in cholestasis resembling that of α‐lipoproteins in a patient with familial deficiency of the enzyme lecithin cholesterol acyl transferase. With anti‐β‐lipoprotein an absence of pre‐β‐lipoproteins with normal electropho‐retic mobility was noted in crossed immunoelectrophoresis. These observations indicate that false values may be obtained with the use of electroimmunoassay for the determination of lipoprotein concentrations in cholestasis. A method employing crossed immunoelectrophoresis with an intermediate gel for the quantitative determination of lipoprotein X is proposed.
Immunoelectrophoretic Quantitation of Maternal Serum Human Placental Lactogen Hormone and Alpha‐Foetoprotein in the Same Electrophoretic Run
Authors: NøRGAARD‐PEDERSEN B. , GæDE P.
Published: February 2009

A technique for quantitative determination of human placental lactogen (HPL) hormone and alpha‐foetoprotein (AFP) in serum or plasma in the same rocket immunoelectrophoresis is described. The HPL rockets are stained with Coomassie brilliant blue and the AFP rockets are visualized by auto‐radiography or by a spark chamber technique. Sixty‐seven HPL‐AFP estimates were made on different sera from 21 women with twin pregnancies. Both the HPL and AFP concentrations were higher in serum from twin pregnancies than in serum from single pregnancies. The diagnostic use of the combined HPL‐AFP assay, especially in the early diagnosis of twin pregnancies and of neural tube malformations, is discussed.
Identification of Human Serum Proteins by Polyacrylamide Gel Electrophoresis Combined with Quantitative Immunoelectrophoretic Methods
Authors: LOFT H.
Published: February 2009

Identification of individual bands in polyacrylamide gels was performed by crossed‐line immunoelectrophoresis in agarose gel secondary to polyacrylamide gel electrophoresis. Line absorption and rocket‐line immunoelectrophoresis were used for identification. Secondly it was shown that incubation of the serum sample with sodium dodecyl sulphate (SDS) does not distort the immunoelectrophoresis pattern, apart from the lipoproteins, provided that the SDS is removed from the polyacrylamide by washing before the immunoelectrophoresis. Simultaneous estimation of apparent molecular weights and identification of the proteins can thus be made.
Detection of Biospecific Interaction during the First Dimension Electrophoresis in Crossed Immunoelectrophoresis*
Authors: BøG‐HANSEN T. C. , BJERRUM O. J. , RAMLAU J.
Published: February 2009

Lectins were included in the first dimension gel in crossed immunoelectrophoresis of human serum proteins. Con A resulted in a retardation of most glycoproteins during the first electrophoresis and heterogeneous forms of individual glycoproteins could be detected. Ulex europeus lectin gave most of the proteins a higher migration velocity. Pokeweed mitogen did not interact with serum glycoproteins, but it did with serum lipoproteins. The method allows a close study of interacting components, determination of binding specificities, and detection of minor interacting components, including heterogeneous molecular forms.
Normally Occurring Precipitating Antibodies against Pseudomonas aeruginosa
Authors: HøIBY N.
Published: February 2009

By means of crossed immunoelectrophoresis with intermediate gel the occurrence of precipitating antibodies against Pseudomonas aeruginosa was studied in sera from 151 'normal' persons. A complex Pseudomonas aeruginosa antigen preparation and a corresponding rabbit antiserum were used as a reference system. Fifty‐four per cent of the sera contained precipitating antibodies against Pseudomonas aeruginosa, and the prevalence of these antibodies increased with age. The specificity and titre of the precipitins were established in relation to the reference system, and the human Pseudomonas precipitins could in this way be referred to 2 Pseudomonas aeruginosa antigens that have been shown to cross‐react with antigens from many other bacterial species. The precipitins were low‐titred and could be absorbed with antigens from 7 other bacterial species. The implications of the results are discussed.
Separation and Identification of Herpes Simplex Virus Antigens by Ion‐Exchange Chromatography and Fused Rocket Immunoelectrophoresis
Authors: VESTERGAARD B. F. , GRAUBALLE P. C.
Published: February 2009

Crude extracts of virus‐infected cells labelled with radioactive isotopes were fractionated by ion‐exchange chromatography. Virus‐specific antigens present in these fractions could be detected by fused rocket immunoelectrophoresis. It was possible to measure the quantitative distribution of 5 HSV type 1 and 10 HSV type 2 virus antigens among 60 fractions. The fractionated antigens were identified by crossed immunoelectrophoresis, using unlabelled reference antigens in the same electrophoretic run. Comparison between the Coomassie brilliant blue picture and the autoradiographic picture of the same plates identified the radioactive‐labelled antigens in relation to the reference antigen scheme.
Purification of Human IgE and Rabbit Anti‐Human IgE
Authors: LøWENSTEIN HENNING , WEEKE BENT
Published: February 2009

Summary. Human IgE from serum containing 60 μg ml‐l was isolated and purified using a combination of gel chromatography, cation exchange chromatography, anion exchange chromatography, and affinity chromatography using con A‐Sepharose®. 1. Anti‐IgE was produced by immunizing rabbits with the IgE‐containing fraction. Specific anti‐IgE was obtained from the isolated rabbit immunoglobulins after absorption of the antibodies against non‐IgE serum proteins on Sepharose 4B, to which was coupled normal human serum enriched withχ and λ chains and a commercial anti‐IgE. 2. The IgE‐containing fraction was coupled to cyanogen bromide activated agarose and used for the production of immunoabsorbed rabbit anti‐human IgE, which was then labelled with 126I for use in RAST and CRIE.
In Vitro Reactivity of Human Lymphocytes after Cryopreservation Using a Programmed Cooling Device
Authors: DU BOIS M. J. G. J. , SCHELLEKENS P. TH. A. , DE WIT J. J. F. M. , EIJSVOOGEL V. P.
Published: February 2009

Abstract. In this article a technique is described for the cryopreservation of human lymphocytes employing a programmed cooling device. A preset temperature curve was used which compensated for the heat exchange during crystallization. The influence of this method of lymphocyte preservation on a variety of the in vitro functions of these cells was evaluated by means such as the lymphocyte transformation test, CML and investigations for lymphocyte membrane markers. All the lymphocyte functions that were tested were found to be retained in full.
Fractionation of Human Blood Lymphocytes on Helix pomatia A Haemagglutinin Coupled to Sepharose Beads
Authors: HELLSTRÖM U. , HAMMARSTRÖM S. , DILLNER M.‐L. , PERLMANN H. , PERLMANN P.
Published: February 2009

Abstract. Treatment of human blood lymphocytes with neuraminidase has previausly been shown to uncover receptors for the A haemagglutinin of the snail Helix pomatia (HP). Neuraminidase‐treated lymphocytes were now fractionated on columns charged with large Sepharose particles to which HP had been coupled covalently. HP‐receptor negative (HP‐) lymphocytes passed the columns while HP‐receptor positive (HP+) lymphocytes were retained. The latter cells were eluted by addition of the competitive hapten N‐acetyl‐D‐galactosamine (D‐GalNAc). The total yield of cells recovered after fractionation was 60‐80% Surface marker studies indicated that there was no selective loss of any of the major lymphocyte subpopulations. The fraction that passed the columns (fraction I) consisted of approximately 10% of all lymphocytes. It contained ˜ 1% HP+ cells and ˜3% of all lymphocytes forming rosettes with sheep erythrocytes (E+ cells) present before fractionation. 50‐55% of the lymphocytes in this fraction had surface‐bound immunoglobulin (SIg+ cells) and complement receptors (EAC+ cells). Of the SIg+ cells, ˜60% were true B cells while the remaining 40% had IgG adsorbed to their surface. The majority of the B cells were recovered in this fraction. The lymphocytes of this fraction responded poorly to T‐cell mitogen but had an enhanced K‐cell activity to chicken erythrocytes. Elution of the cells retained on the column with 0.1 mg/ml D‐GalNAc gave a fraction II, consisting of, ˜15% of all lymphocytes. This fraction had a mixed composition. The majority of the cells (˜45%) were recovered by subsequent elution with 1.0 mg/ml D‐GalNAc. This fraction III was strongly enriched with HP+ and E+ cells (T cells). About 10% of the HP+ cells in this fraction were SIg+. However, on the majority of these cells this surface‐bound immunoglobulin was probably externally adsorbed IgG. These HP+‐SIg+ cells were also EAC+ and had Fc receptors, as shown by rosette formation with IgG‐coated bovine erythrocytes. The lymphocytes of fraction III responded most strongly to T‐cell mitogen while their K‐cell activity was weak.
A Case of Ovarian Endodermal Sinus Tumour with Recurrence of a Large Benign Teratoma
Authors: KNUDSEN J. , BERTHELSEN A. , ALBRECHTSEN R. , NØRGAARD‐PEDERSEN B.
Published: February 2009

A 14 year old girl was operated for a left side ovarian tumour (weight: 1900 g). Histological examination: endodermal sinus tumour, combined with embryonal carcinoma and dysgerminoma. Serum‐alpha‐fetoprotein (AFP) was highly elevated (2630 μg/l), and serum human chorionic gonadotropin (hCG) was slightly elevated (100 IU/l), but 2 1/2 months after operation the AFP concentration was less than 20 μg/l, and hCG less than 15 IU/l. After 2 years of intermittent treatment with combination chemotherapy (VAMBLE) the patient developed a left side abdominal tumour (weight: 4600 g). Histological examination: Benign teratoma. There was no sign of recurrence of the malignant disease, and serum AFP and hCG are still normal.
Immunofluorescent Demonstration of Alpha‐fetoprotein in Tissue Culture from Human Endodermal Sinus Tumors, Rat Yolk Sac Carcinoma and Fetal Rat Yolk Sac
Authors: ALBRECHTSEN R. , HIRAI H. , LINDER D. , NØRGAARD‐PEDERSEN B. , WEWER U.
Published: February 2009

An in vitro model for study of AFP (alpha‐fetoprotein) synthesis is presented using immunofluorescent microscopic examination of cultured cells from rat yolk sac, experimental rat yolk sac carcinoma and human endodermal sinus tumors (EST).

The presence of positive staining for AFP in all cases by use of specific rabbit antisera against rat and human AFP supports the concept that AFP synthesis in germ cell tumors is confined to the yolk sac endoderm. The variation of positive staining cells due to a variable protein synthesis is discussed.
Rat Yolk Sac Tumor: Biological Aspects and Microheterogeneity of AFP
Authors: SAKASHITA S. , TSUKADA Y. , HIRAI H. , TSUJI I.
Published: February 2009

α‐Fetoprotein (AFP)‐producing yolk sac tumors were established in rats as transplantable tumor lines growing in either solid or ascitic form. AFP levels in the ascites and in the serum of recipients were quite high, but higher in castrated female rats than male rats. The tumor cells were cultured also in vitro and showed synthesis of AFP and albumin. AFP produced by the yolk sac tumors was investigated comparatively with AFP produced by a hepatoma. There was no remarkable difference between two AFPs in behavior in gel filtration; however, AFP of yolk sac tumors migrated more slowly than that of hepatoma in immunoelectrophoresis. However, the mobilities of two AFP preparations became same after desialization by neuraminidase treatment. This fact suggests that AFP of yolk sac tumors contains sialic acids in smaller amount than that of hepatomas.
Immunofluorescent Localization of Alpha‐fetoprotein Synthesis in the Endodermal Sinus of Rat Placenta
Authors: ALBRECHTSEN R. , NØRGAARD‐PEDERSEN B.
Published: February 2009

Specific immunofluorescence staining for AFP of the rat yolk sac showed that the synthesis was confined to the visceral layer of the yolk sac endoderm. The AFP synthesis took place not only in the extraplacental yolk sac as demonstrated by Gitlin in 1967, but also in the endodermal sinus structures within the placenta. No immunofluorescence staining was found of Reichert's membrane close to the parietal layer of the yolk sac. These findings are in accordance with the earlier demonstrated AFP synthesis in the visceral layer of the endodermal sinus structures in the tumors.
Hormonal Control and Putative Cell Cycle Dependency of AFP Production: Further Observations in vivo and in vitro
Authors: BÉLANGER L. , HAMEL D. , DUFOUR D. , GUILLOUZO A. , CHIU J. F.
Published: February 2009

We describe the effect of hormones on AFP production in neonatal rats, in primary fetal rat hepatocyte cultures, in adult animals with liver injuries, and in tumor cell lines established in vitro. Preliminary data of studies on the putative cell cycle dependency of AFP production are also presented. Suppression of AFP by glucocorticoid hormones emerges as a most powerful system to study liver cell differentiation through AFP regulation.
Light and Electron Microscope Immunolocalization of Alpha‐fetoprotein in Rat Liver Cells in vivo and in vitro
Authors: GUILLOUZO A. , BÉLANGER L. , BEAUMONT C. , BRIGGS R. , CHIU J. F. , FLEISCHER B. , FLEISCHER S. , VALET J. P.
Published: February 2009

In neonatal rat liver, AFP is localized only in typical hepatocytes. Their lobular distribution changes throughout the neonatal period. Some AFP+ cells also contain albumin. Less than 5% of AFP+ cells incorporate 3H‐thymidine (2‐hour pulse). Injections of dexamethasone suppress AFP positivity but not albumin positivity nor 3H‐thymidine incorporation. AFP is also localized in typical hepatocytes in newborn rat isolated liver cells or liver explants in culture. During the early phase of AFP induction in rats, by mDAB feeding, AFP is detected in cells smaller than the normal hepatocytes and preferentially situated in periportal areas of the liver.

Electron microscope immunoperoxidase localizations in newborn rat liver show that AFP is present on bound ribosomes, in the lumina of rough and smooth endoplasmic reticulum, and in the Golgi apparatus. Direct AFP measurement on isolated organelles confirmed this distribution. It indicates a synthesis and secretion pattern similar to that of albumin.
Immunochemical Characteristics of Concanavalin‐A Reactive and Non‐reactive Alpha‐fetoprotein
Authors: DAMBUYANT C. , SIZARET Ph. , MARTEL N.
Published: February 2009

The existence of two alpha‐fetoproteins, one which does and the other which does not bind the concanavalin A, was confirmed by affinity chromatography. The non‐binding alpha‐fetoprotein represented about 2.3% of the total and the binding alpha‐fetoprotein was constituted by a series of fractions with different avidities. The electrophoretic mobility of fractions which bound was greater and this was due to differences in charge, not in molecular weight; on the other hand binding and non‐binding alpha‐fetoproteins were identical when compared by gel diffusion; the slopes of dose‐response curves obtained by the radioimmunoassay with twelve antisera were steeper for binding fractions but differences with non‐binding alpha‐fetoprotein were not significant. Alpha‐mannopyranosyl or sterically identical determinants of alpha‐fetoprotein through which the binding to the concanavalin A occurs are different from alpha‐fetoprotein antigenic sites; both can be weakly masked by sialic acid.
Low Molecular Weight CEA, Isolation and Characterization
Authors: HEDIN A. , HAMMARSTRÖOM S. , SVENBERG T. , SUNDBLAD G.
Published: February 2009

A low molecular weight CEA‐fraction (CEAlow) was purified from liver metastases of colorectal cancer. The following purification sequence was used: homogenization—PCA extraction—ion‐exchange chromatography— Con A affinity chromatography—gel filtration on Sephadex G‐200 (x2)—affinity chromatography on specific anti‐CEA immunoadsorbent—gel filtration (G‐200).

CEAlow was homogeneous on gel filtration in 6M guanidine‐HCl and gave a single band on SDS‐PAGE. CEAlow and CEA gave a reaction of identity in immunodiffusion with 7/9 monkey anti‐CEA sera. 2/9 monkey anti‐CEA sera showed a weak spur, with CEA spurring over CEAlow. CEAlow gave a reaction of partial identity with NCA and BGP‐1 with unabsorbed rabbit and sheep anti‐CEA sera. CEAlow consisted of a single polypeptide chain and had a molecular weight of 125,000 as compared to 175,000 ± 8,000 for CEA (SDS‐PAGE). CEA and CEAlow contained the same sugars and aminoacids in approximately the same molar proportions. However the carbohydrate content of CEAlow was lower (30–40% as compared to 45–50% for CEA). The peptide relatedness (expressed in SΔQ‐units) of CEA, CEAlow, NCA and BGP‐I was compared. CEAlow was most closely related to CEA (SΔQ‐values of 5–7) followed by NCA (SΔQ values of 12–17) and BGP‐I (SΔQ‐values of 35–40).

It is clear from these studies that CEAlow is closely related to CEA but contains less carbohydrate. Whether the polypeptide chain is shorter remains to be established.
Antigenic Determinants of Carcinoembryonic Antigen
Authors: ORMEROD M. G.
Published: February 2009

Radioimmunoassays for carcinoembryonic antigen (CEA) have been established using a variety of different antisera raised in goat, monkey, rabbit, guinea pig and rat. The extent to which different antigens compete for antibodies in the sera has been estimated. The antigens used were: CEA, asialo CEA, Smith‐degraded CEA (50% carbohydrate removed), CEA treated with an extract of Trichomonas foetus (70% carbohydrate removed), reduced‐alkylated CEA (disulphide bonds ruptured), a peptic digest of CEA and CEX. The results demonstrated that there are at least five chemically distinct antigenic determinants: three are on protein—one, sensitive to conformation, a second, sensitive to conformation and to periodate, a third, insensitive to conformation—and two on carbohydrate, one common to CEA and CEX and another on CEA alone.
Placental Alkaline Phosphatase Appearance in Murine Lymphomas
Authors: LUMB J. R. , FLOYD R. A.
Published: February 2009

Alkaline phosphatase (APase) has been shown to have a membrane‐bound localization in the murine fetal thymus, murine thymic lymphoma and in areas of lymphoproliferation of the adult spleen. A detailed biochemical comparison of the lymphoma APase with the fetal thymus, placenta, and spleen APases was performed. The parameters investigated were pH optimum, activation, inhibition, heat inactivation and substrate ratio with the substrate α‐naphthyl phosphate. All four murine APase activities have a pH optimum of 10 and were activated by magnesium and nucleophilic buffers to the same extent. The APase activities tested were all inhibited by ethylenediamine tetraacetic acid, L‐phenylalanine and L‐homoarginine at the same levels, approximately 97%, 8% and 81%, respectively. Substrate ratios indicate that all four APases hydrolyzed both p‐nitrophenyl phosphate and α‐naphthyl phosphate at the same rate. These data lead one to conclude that the APase of murine thymic lymphoma may represent a cell membrane carcinofetal enzyme.
Occurrence in Normal Human Testis of a Heat‐stable Alkaline Phosphatase Similar to a Rare Variant of Placental Isoenzyme
Authors: CHANG C.‐H. , ANGELLIS D. , FISHMAN W. H.
Published: February 2009

The possibility that testicular germ cells are ‘totipotential’ in expressing early developmental gene products was examined in the case of placenta‐specific heat‐stable alkaline phosphatase. In each of the 11 adult testes studied about 0.3–4.6% of the total alkaline phosphatase activity was heat‐stable and L‐phenylalanine‐sensitive but L‐homoarginine‐insensitive. However, the testicular heat‐stable enzyme was more susceptible to inhibition by L‐leucine and EDTA than the normal placental and intestinal enzymes, thus resembling the so‐called ‘D‐variant’ of placental alkaline phosphatase. By antibody‐directed enzyme inhibition tests the testicular heat‐stable enzyme crossreacted completely with normal placental enzyme but clearly distinguished itself from a heat‐stable component of normal intestinal enzyme. Thus, a minute amount of placental alkaline phosphatase D‐variant is synthesized in testis, indicating that the gene for elaborating this placental protein is probably already active in the germ cells of testis. The high incidence of this protein in cancers of testis and ovary is probably due to the increased production of this protein by active gonadal genes.
Serial Measurements of Pregnancy‐specific β1‐glycoprotein (β1SP1) in Patients with Trophoblastic Disease
Authors: SEARLE F. , BAGSHAWE K. D. , DENT J. , LEAKE B. A.
Published: February 2009

A radioimmunoassay for pregnancy‐specific‐β1‐glycoprotein (Bohn. 1971) has been established with a limit of detection of 2 μMg/litre in serum. The assay has been used to measure serial levels of pregnancy‐specific‐β1‐glycoprotein (β1SP1) in the serum of patients with choriocarcinoma and teratoma for comparison with measurements of the β‐subunit of human chorionic gonadotrophin. The value of the assay for β1SP1, in the management of these patients is discussed.
Immunosuppressive Activity of α 2H‐‘isoferritin’ as Compared to Normal Crystallized Ferritin
Authors: BUFFE D. , RIMBAUT C. , ERARD D. , TILZ G. P.
Published: February 2009

The suppressive activity of α 2H‐isoferritin was studied in vivo and in vitro.

α 2H‐isoferritin inhibits the hemolysin synthesis in mice and the T independent and T dependent antibodies induced in vitro.

The immune response of normal human lymphocytes is suppressed as shown by lymphoblastic transformation induced by mitogen (PHA), antigens (tuberculin and Candidin), mixed lymphocyte cultures.

The inhibitory effect is proportional to the quantity of added protein. The normal ferritin has no effect. The role of the carbohydrate component is discussed.
Effectiveness of Rex shunt in children with portal hypertension following liver transplantation or with primary portal hypertension
Authors: Krebs‐Schmitt D. , Briem‐Richter A. , Grabhorn E. , Burdelski M. , Helmke K. , Broering D. C. , Ganschow R.
Published: February 2009

Abstract:  Portal vein thrombosis can occur as a result of primary anomalies, after liver transplantation, and for other reasons. It may result in severe complications secondary to portal hypertension, such as bleeding from esophageal or gastric varices, hypersplenism, or impaired somatic growth. In this retrospective study, we analyzed the outcome of 25 children who underwent a Rex shunt procedure. The following venous grafts were used as the shunt: the autologous internal or external jugular vein (n = 17) or a cryopreserved graft (n = 5); in three patients the umbilical vein was recanalized. The median follow up time was 109 months (range 18 days–146 months). The best results were achieved in patients in whom an autologous jugular vein segment was used as a vascular graft for the Rex shunt (shunt patency of 88%). In patients with a functioning shunt no further lower or upper gastrointestinal bleeding occurred. And in the entire study population hypersplenism syndrome improved after surgery. In our large cohort of pediatric patients, the Rex shunt has shown to be an effective method to eliminate portal hypertension and to revascularize the liver and thereby prevents the possible consequences of long‐term portosystemic shunting.
ELECTIVE TWIN REDUCTIONS: EVIDENCE AND ETHICS
Authors: MCCLIMANS LEAH
Published: February 2009

Twelve years ago the British media got wind of a London gynecologist who performed an elective reduction on a twin pregnancy reducing it to a singleton. Perhaps not surprisingly, opinion on the moral status of twin reductions was divided. But in the last few years new evidence regarding the medical risks of twin pregnancies has emerged, suggesting that twin reductions are relevantly similar to the reductions performed on high‐end multi‐fetal pregnancies. This evidence has appeared to resolve the moral debate.

In this paper I look at the role of clinical evidence in medical ethics. In particular I examine the role of clinical evidence in determining what counts as a significant harm or risk. First, I challenge the extent to which these empirical claims are descriptive, suggesting instead that the evidence is to some degree normative in character. Second, I question whether such empirical claims should count as evidence for what are essentially difficult ethical decisions – a role they appear to play in the case of elective reductions. I will argue that they should not, primarily because the value‐laden nature of this evidence conceals much of what is ethically at stake. It is important to recognize that empirical evidence cannot be a substitute for ethical deliberation.
Restrictive Behaviour Management Procedures with People with Intellectual Disabilities who Require Dental Treatment
Authors: Newton J. T.
Published: February 2009

Background  Dental disease is more common among people with intellectual disabilities than in the general population. Improvements in oral health require individuals to engage in daily oral hygiene and regular visits to a dental practitioner; both may be challenging for the individual with intellectual impairment.

Materials and Methods  A review of policies relating to behaviour management and physical restraint for individuals with intellectual disabilities and challenging behaviours was undertaken. Published studies of behaviour management in individuals with intellectual disabilities attending for dental treatment were reviewed. Reference to studies of children with average IQ and other populations were made where appropriate.

Results  There is little published evidence regarding behaviour management for people with intellectual disabilities who require dental treatment. Current policies place great emphasis on pharmacological management and restrictive behaviour management techniques. There is a paucity of studies which have employed a functional analysis framework. There are few incentives for dentists to implement positive approaches to behaviour management as current systems of payment reward the completion of numbers of treatments and thus there is an incentive to complete treatments quickly rather than to spend time with patients.

Conclusions  There is a need for research addressing the behaviour management of individuals with intellectual disabilities who require dental treatment, particularly the efficacy of non‐restrictive techniques and the provision of incentives that reward the use of positive behaviour management among dentists. The use of restrictive techniques should be carefully monitored for compliance with best practice guidelines.
Restraint Safety: an Analysis of Injuries Related to Restraint of People with Intellectual Disabilities
Authors: Williams Don E.
Published: February 2009

Background  There is little research on the safety of the various types of restraint commonly used with individuals with intellectual disabilities who exhibit severely aggressive or self‐injurious behaviour.

Method  This study analysed the use of restraint with 209 individuals with intellectual disabilities over a 12‐month period.

Results  Planned restraint, the use of restraint as a component of a behaviour treatment programme (i.e. planned personal or planned mechanical restraint) was safer than crisis‐intervention restraint (emergency personal or emergency mechanical). The overall rate of injuries during restraint was 0.46 injuries per hundred restraints. Restraint was applied 99.54% of the time without injury.

Conclusion  Restraint was relatively safe and safer than reported in one other study. Planned restraint was safer than emergency restraint. The additional training and programme development associated with planned restraint may have contributed to the greater safety of planned restraint. Due to limited empirical data, restraint safety has yet to be established and this study suggests that restraint should be decreased and closely monitored.
The Effects of Non‐Contingent Self‐Restraint on Self‐Injury
Authors: Kerth Denise Marzullo , Progar Patrick R. , Morales Sabrina
Published: February 2009

Background  Self‐restraint is a pervasive phenomenon among individuals who engage in self‐injurious behaviour (SIB).

Materials and Methods  The present study examined the use of clothing as a socially acceptable alternative to self‐restraint to reduce SIB and other topographies of self‐restraint in an adolescent diagnosed with autism. Two separate functional analyses were conducted for SIB prior to the self‐restraint evaluation. A reversal design was then used to evaluate the effect of non‐contingent access to a hooded sweatshirt on rates of self‐restraint and SIB.

Results  Both functional analyses suggested that SIB was maintained by access to tangible items, attention, and perhaps also by escape from demands. The results of the self‐restraint evaluation indicated that when access to a hooded sweatshirt was provided, rates of other topographies of self‐restraint dropped to zero and rates of SIB were reduced by 54% from baseline levels.

Conclusions  These results highlight the idiosyncratic relationship between SIB and self‐restraint and suggest that systematic evaluations of this relationship may lead to reductions in both behaviours.
Evaluation of Timeout Programs through Phased Withdrawal
Authors: Iwata Brian A. , Rolider Natalie U. , Dozier Claudia L.
Published: February 2009

Background  Intrusive interventions, once implemented, may remain in place longer than necessary unless their continued effectiveness is evaluated.

Method  We conducted a phased timeout‐evaluation for 34 individuals with developmental disabilities who received services from the same agency and whose treatment programmes contained timeout for aggression and/or property destruction. After establishing baseline rates of problem behaviour (with timeout in effect), we removed timeout from all programmes through a gradual process that occurred over a 1‐year period.

Results  Rates of problem behaviour decreased (or did not increase) for 21 individuals following the removal of timeout. Rates of problem behaviour increased for 13 individuals, but alternative procedures that were less intrusive than timeout were implemented for 10 individuals. Timeout was reinstated for the remaining three individuals and was associated with reductions in problem behaviour.

Conclusions  The phased timeout evaluation resulted in successful removal of 91% (31 of 34) of the original timeout programmes.
Effects of gustatory stimulants of salivary secretion on salivary pH and flow: a randomized controlled trial
Authors: Da Mata ADSP , Da Silva Marques DN , Silveira JML , Marques JROF , De Melo Campos Felino ET , Guilherme NFRPM
Published: February 2009

Objectives:  To compare salivary pH changes and stimulation efficacy of two different gustatory stimulants of salivary secretion (GSSS).

Setting:  Portuguese Dental Faculty Clinic.

Design:  Double blind randomized controlled trial.

Subjects:  One hundred and twenty volunteers were randomized to two intervention groups. Sample sized was calculated using an alpha error of 0.05 and a beta of 0.20.

Materials and methods:  Participants were randomly assigned to receive a new gustatory stimulant of secretory secretion containing a weaker malic acid, fluoride and xylitol or a traditionally citric acid‐based one. Saliva collection was obtained by established methods at different times. The salivary pH of the samples was determined with a pH meter and a microelectrode.

Main outcome measures:  Salivary pH variations and counts of subjects with pH below 5.5 for over 1 min and stimulated salivary flow were the main outcome measures.

Results:  Both GSSS significantly stimulated salivary output without significant differences between the two groups. The new gustatory stimulant of salivary secretion presented a risk reduction of 80 ± 10.6% (95% CI) when compared with the traditional one.

Conclusions:  Gustatory stimulants of salivary secretion with fluoride, xylitol and lower acid content maintain similar salivary stimulation capacity while reducing significantly the dental erosion predictive potential.