bluuebunny/crossref_metadata_2025 huggingface dataset:
Altered early infant gut microbiota in children developing allergy up to 5 years of age
Authors: Sjögren Y. M. , Jenmalm M. C. , Böttcher M. F. , Björkstén B. , Sverremark‐Ekström E.
Published: February 2009

Background
Early colonization with bifidobacteria and lactobacilli is postulated to protect children from allergy, while Clostridium (C.) difficile colonization might be associated with allergic disease. Previous studies of infant gut microbiota in relation to subsequent allergy development have mostly employed culture‐dependent techniques, studied genera of bacteria and the follow‐up period was limited to 2 years.

Objective
To relate gut microbiota in early infancy, notably bifidobacteria and lactobacilli at species level, to allergy development during the first 5 years of life and study if environmental factors influence the early infant gut microbiota.

Methods
Fecal samples were collected at 1 week, 1 month and 2 months after birth from 47 Swedish infants, followed prospectively to 5 years of age. Bacterial DNA was analysed with real‐time PCR and related to allergy development, family size as well as endotoxin and Fel d 1 levels in house dust samples. Primers binding to C. difficile, four species of bifidobacteria, two lactobacilli groups and Bacteroides fragilis were used. Children regarded as allergic manifested allergic symptoms and were skin prick test positive during their first 5 years while non‐allergic children were neither.

Results
Children who developed allergy were significantly less often colonized with lactobacilli group I (Lactobacillus (L.) rhamnosus, L. casei, L. paracasei), Bifidobacterium adolescentis and C. difficile during their first 2 months. Infants colonized with several Bifidobacterium species had been exposed to higher amounts of endotoxin and grew up in larger families than infants harbouring few species.

Conclusion
A more diverse gut microbiota early in life might prevent allergy development and may be related to the previously suggested inverse relationship between allergy, family size and endotoxin exposure.
Interleukin‐12 is sufficient to promote antigen‐independent interferon‐γ production by CD8 T cells in old mice
Authors: Rottinghaus Erin K. , Vesosky Bridget , Turner Joanne
Published: February 2009

Numerous functional defects have been identified in naive T cells from aged mice, including deficiencies in proliferation, cytokine production and signal transduction. It is well documented that the ratio of naïve to memory T cells significantly decreases with age resulting in the majority of T cells from aged hosts expressing activated/memory T‐cell markers (CD44hi), yet it is unclear whether T cells with a CD44hi phenotype in aged hosts are functionally equivalent to T cells with a similar phenotype in young hosts. We have identified a population of CD44hi CD8 T cells in old mice that are capable of secreting interferon‐γ (IFN‐γ) in response to interleukin‐12 (IL‐12) stimulation. This occurred in the absence of T‐cell receptor engagement, a function that was not observed in CD8 T cells from young mice. This phenotype was associated with increased IL‐12 receptor β2 gene expression and IL‐12 induced signal transducer and activator of transcription 4 (STAT‐4) activation, even when CD8 T‐cell numbers from young and old mice were normalized for CD44hi expression. Furthermore, we demonstrate that IL‐12‐induced STAT‐4 activation was required for T helper type 1 (Th1) cytokine‐induced IFN‐γ production in CD8 T cells. These data illustrate that old mice possess a specialized subset of CD44hi CD8 T cells with an enhanced responsiveness to IL‐12, enabling these cells to produce substantial amounts of IFN‐γ in response to Th1 cytokine stimulation. We have therefore identified a functional difference in the populations of CD44hi CD8 T cells from young and old mice, and believe that understanding age‐associated immunological changes is essential for helping the elderly combat deadly diseases.
Astrocyte elevated gene‐1 (AEG‐1) is induced by lipopolysaccharide as toll‐like receptor 4 (TLR4) ligand and regulates TLR4 signalling
Authors: Khuda Imtiaz I‐E. , Koide Naoki , Noman Abu S. M. , Dagvadorj Jargalsaikhan , Tumurkhuu Gantsetseg , Naiki Yoshikazu , Komatsu Takayuki , Yoshida Tomoaki , Yokochi Takashi
Published: February 2009

Astrocyte elevated gene‐1 (AEG‐1) is induced by human immunodeficiency virus 1 (HIV‐1) infection and involved in tumour progression, migration and invasion as a nuclear factor‐κB (NF‐κB) ‐dependent gene. The involvement of AEG‐1 on lipopolysaccharide (LPS) ‐induced proinflammatory cytokine production was examined. AEG‐1 was induced via NF‐κB activation in LPS‐stimulated U937 human promonocytic cells. AEG‐1 induced by LPS subsequently regulated NF‐κB activation. The prevention of AEG‐1 expression inhibited LPS‐induced tumour necrosis factor‐α and prostaglandin E2 production. The AEG‐1 activation was not induced by toll‐like receptor ligands other than LPS. Therefore, AEG‐1 was suggested to be a LPS‐responsive gene and involved in LPS‐induced inflammatory response.
Suppression of antitumour protective cytotoxic T lymphocyte responses to a human papillomavirus 16 E7 DNA vaccine by coinjection of interleukin‐12 complementary DNA: involvement of nitric oxide in immune suppression
Authors: Sin Jeong‐Im
Published: February 2009

Interleukin‐12 (IL‐12) has been shown to enhance cellular immunity in vitro and in vivo. The beneficial roles of IL‐12 as a DNA vaccine adjuvant have been commonly observed. Here the impact of IL‐12 complementary DNA (cDNA) as an adjuvant for a human papillomavirus (HPV) type 16 E7 DNA vaccine is investigated in a mouse tumour model. Coinjection of E7 DNA vaccine with IL‐12 cDNA completely suppressed antigen‐specific cytotoxic T‐lymphocyte (CTL) responses, leading to a complete loss of antitumour protection from a tumour cell challenge. In addition, antigen‐specific antibody and T helper cell proliferative responses were also suppressed by IL‐12 cDNA coinjection. This inhibition was observed over different IL‐12 cDNA doses. Furthermore, separate leg injections of IL‐12 and E7 cDNAs suppressed antigen‐specific CTL and tumour protective responses, but not antibody and T helper cell proliferative responses, suggesting different pathways for suppression of these two separate responses. Further knockout animal studies demonstrated that interferon‐γ and nitric oxide are not directly associated with suppression of antigen‐specific antibody responses by IL‐12 cDNA coinjection. However, nitric oxide was found to be involved in suppression of antigen‐specific CTL and tumour protective responses by IL‐12 cDNA coinjection. These data suggest that coinjection of IL‐12 cDNA results in suppression of E7‐specific CTL responses through nitric oxide, leading to a loss of antitumour resistance in this DNA vaccine model. This study further shows that the adjuvant effect of IL‐12 is dependent on the antigen types tested.
A central role for monocytes in Toll‐like receptor‐mediated activation of the vasculature
Authors: Ward Jon R. , Francis Sheila E. , Marsden Luke , Suddason Tesha , Lord Graham M. , Dower Steven K. , Crossman David C. , Sabroe Ian
Published: February 2009

There is increasing evidence that activation of inflammatory responses in a variety of tissues is mediated co‐operatively by the actions of more than one cell type. In particular, the monocyte has been implicated as a potentially important cell in the initiation of inflammatory responses to Toll‐like receptor (TLR)‐activating signals. To determine the potential for monocyte‐regulated activation of tissue cells to underpin inflammatory responses in the vasculature, we established cocultures of primary human endothelial cells and monocytes and dissected the inflammatory responses of these systems following activation with TLR agonists. We observed that effective activation of inflammatory responses required bidirectional signalling between the monocyte and the tissue cell. Activation of cocultures was dependent on interleukin‐1 (IL‐1). Although monocyte‐mediated IL‐1β production was crucial to the activation of cocultures, TLR specificity to these responses was also provided by the endothelial cells, which served to regulate the signalling of the monocytes. TLR4‐induced IL‐1β production by monocytes was increased by TLR4‐dependent endothelial activation in coculture, and was associated with increased monocyte CD14 expression. Activation of this inflammatory network also supported the potential for downstream monocyte‐dependent T helper type 17 activation. These data define co‐operative networks regulating inflammatory responses to TLR agonists, identify points amenable to targeting for the amelioration of vascular inflammation, and offer the potential to modify atherosclerotic plaque instability after a severe infection.
Effects of buccal bi‐cortical anchorages on primary stability of dental implants: a numerical approach of natural frequency analysis
Authors: WANG K. , LI D. H. , GUO J. F. , LIU B. L. , SHI S. Q.
Published: February 2009

Summary  The purpose of this study was to investigate influences of buccal bi‐cortical anchorages on natural frequency (NF) values of dental implants in different diameters utilizing the three‐dimensional finite element method. Three degrees of buccal bi‐cortical engagements were generated in D2 and D3 bone quality models, which were 0‐mm engagement (i.e. implants just had contact with the buccal cortex), 0·5‐mm (i.e. implants were penetrated into the buccal cortex by 0·5 mm) and 1·0‐mm engagement, while only 0‐ and 0·5‐mm engagement were simulated in D4 bone models. The uni‐cortical engagement was set as the control. By the modal analysis, NF values of bending and axial vibration mode were computed as a function of different bi‐cortical engagements. The results showed that buccal bi‐cortical anchorages significantly enhanced bending and axial NF values. The increasing rates resulting from 0·5‐mm engagement ranged from 10·5 to 42·3%, with a mean of 24·3%. From 0‐ to 0·5‐mm engagement, the NF values maintained an increasing trend, and from 0·5‐ to 1·0‐mm engagement, the values levelled off or even decreased. In 0·5‐ and 1·0‐mm engagement models, increasing implant diameter resulted in small increases of NF values. In conclusion, buccal bi‐cortical anchorages could significantly increase both bending and axial NF values of dental implants, but extra‐buccal cortical bone engagement could not produce considerable incremental increases of NF values as anticipated. Increasing implant diameter could result in limited increases of NF values in case of implants being bi‐cortically anchored.
Agreement between patient and proxy assessments of oral health‐related quality of life after stroke: an observational longitudinal study
Authors: MCGRATH C. , MCMILLAN A. S. , ZHU H. W. , LI L. S. W.
Published: February 2009

Summary  This study aimed to assess agreement between patient and proxy assessment of patients’ oral health‐related quality of life (OHRQoL) during the acute stroke phase and 6 months after hospital discharge. As part of an observational longitudinal study, 161 consecutive patients hospitalized after stroke and their caregivers (CGs) were monitored during acute stroke and 6 months after hospital discharge. Patient–CG agreement of health‐related quality of life as assessed by the General Oral Health Assessment Index (GOHAI) and Medical Outcomes Study 12‐item Short Form (SF‐12) which comprises physical (PCS‐12) and mental component summaries (MCS‐12) was determined by comparison and correlation analyses. Complete data were available for 65 patients and CGs. Compared with acute stroke, there were improvements in GOHAI and SF‐12 scores from patient’s and CG’s perspective at 6‐month follow‐up. Caregivers overestimated the impact of stroke on MCS‐12 over a period of time (P < 0·01) and underestimated the impact on GOHAI during acute stroke (P < 0·001) and on PCS‐12 6 months later (P < 0·05). Patient–CG agreement was substantial to excellent during acute stroke [intra‐class correlation coefficient (ICC 0·69–0·86)] and fair‐to‐moderate 6 months later (ICC 0·28–0·60). Agreement was adequate between patients’ and CGs’ assessment of patient OHRQoL during acute stroke but it was less reliable 6 months after hospital discharge. These findings suggest that proxy assessment of OHRQoL after stroke is effective in assessing stroke‐related oral impairments that affect life quality without placing additional burden on stroke patients themselves to assess OHRQoL.
Noninvasive diagnosis of liver fibrosis in patients with HIV infection and HCV/HBV co‐infection
Authors: Moreno S. , García‐Samaniego J. , Moreno A. , Ortega E. , Pineda J. A. , Del Romero J. , Tural C. , Von Wichmann M. A. , Berenguer J. , Castro Á. , Espacio R.
Published: February 2009

Summary.  The measurement of fibrosis stage critically affects the identification of the progression of liver disease, the establishment of a prognosis and therapeutic decision making. Liver biopsy has been the single, most useful method to determine the degree of liver fibrosis (LF), but with recognized limitations, mainly associated with its invasiveness. In recent years, alternative noninvasive methods have been developed, including imaging methods, such as transient elastometry, and assays based on serum biomarkers. This article reviews the available studies evaluating the value of various noninvasive methods for the assessment of LF in patients with HIV‐infection and HBV/HCV co‐infection, and makes recommendations on how to best use and combine them in clinical practice.
Chronic hepatitis C and persistent occult hepatitis C virus infection are characterized by distinct immune cell cytokine expression profiles
Authors: Pham T. N. Q. , Mercer S. E. , Michalak T. I.
Published: February 2009

Summary.  Hepatitis C virus (HCV) replicates in immune cells in both chronic hepatitis C (CHC) and occult HCV infection, but the extent of virus replication in this compartment in these opposing infection forms varies greatly. It was unknown whether this could be linked to HCV genotype or to differences in host gene expression shaping the immune response, and whether HCV replication in immune cells is sensitive to endogenous antiviral cytokines. In this study, we uncovered that significantly greater HCV load in peripheral blood mononuclear cells (PBMC), but not in plasma, coincided with HCV genotypes 2 and 3 in CHC, but with genotype 1 in residual occult infection after clinical resolution of hepatitis C. Moreover, PBMC from individuals with occult infection transcribed significantly greater levels of IFN‐α, IFN‐γ and TNF‐α, but less interleukin (IL)‐10 than those from CHC. In CHC, PBMC with low HCV load expressed significantly more IFN‐γ but less IL‐12 than did cells with high virus content. In occult infection, HCV RNA detection in PBMC was associated with much lower IFN‐α and IL‐12 expression. Further, HCV replication in T lymphocytes could be completely eliminated by activation of endogenous IFN‐γ in CHC, but of IFN‐α in occult infection. In conclusion, CHC and persistent occult HCV infection are characterized by clearly different profiles of antiviral cytokine response in circulating immune cells which are also different from those of healthy individuals. Higher expression of IL‐10, combined with lower transcription of IFN‐α, IFN‐γ and TNF‐α, is associated with a more robust HCV replication in immune cells.
Does fluvastatin favour HCV replication in vivo? A pilot study on HIV–HCV coinfected patients
Authors: Milazzo L. , Meroni L. , Galazzi M. , Cesari M. , Caramma I. , Marchetti G. , Galli M. , Antinori S.
Published: February 2009

Summary.  Fluvastatin showed anti‐hepatitis C virus (HCV) activity in vitro, through the inhibition of geranylgeranylation of cellular proteins, and a synergistic effect with interferon (IFN)‐α. Nevertheless statins up‐regulate low‐density lipoprotein (LDL) receptor, required for HCV cell entry, and the closely related scavenger receptors SRBI and CD36; moreover they reduce class II major histocompatibility complex expression on antigen presenting cell, modulating T‐cell activation. In vivo LDL levels have been identified as prognostic indicator of sustained viral response to IFN in patients with HCV infection, suggesting that lipid‐lowering agents might conversely favour HCV entry into the hepatocytes and translate into higher viral replication. We evaluated the effect of fluvastatin on HCV‐RNA levels, CD36 expression and T‐cell homeostasis in HCV‐RNA positive patients. HCV‐RNA was measured at baseline and after 4 weeks in 42 HCV/HIV‐1 co‐infected patients, randomized to receive either fluvastatin 80 mg qd or no treatment. CD36 expression and markers of T‐cell activation were evaluated by means of flow cytometry. Plasma interleukin (IL)‐10, IFN‐γ and IL‐7 were measured by ELISA. Serum cholesterol and LDL decreased significantly in the treatment group (P = 0.0001 and 0.01, respectively). Surprisingly a significant increase of HCV‐RNA levels was seen after 4 weeks of fluvastatin (P = 0.03). The percentages of naive/activated/apoptotic cells and CD36 expression remained unchanged. Fluvastatin did not inhibit HCV‐RNA replication in vivo; conversely we observed a significant increase of HCV‐RNA levels. CD36 expression on monocytes were not up‐regulated by statins as previously reported in vitro. The correlation between HCV infectivity, oxidized‐LDL receptor and statins in HCV infection need further evaluation.
Clinical effects of STW 5 (Iberogast®) are not based on acceleration of gastric emptying in patients with functional dyspepsia and gastroparesis
Authors: Braden B. , Caspary W. , Börner N. , Vinson B. , Schneider A. R. J.
Published: February 2009

Abstract  STW 5, a herbal extract, is effective for the treatment of symptoms in patients with functional dyspepsia (FD). However, its mode of action is still unclear and a modulation of gastric motility is hypothesized. This multicentre, placebo‐controlled double‐blind study addressed the question of whether STW 5 accelerates gastric emptying in patients with FD and gastroparesis. One‐hundred and three patients diagnosed with FD were randomly assigned to a treatment with either STW 5 or a liquid placebo for 28 days. The primary end point of the study was a change of a validated gastrointestinal symptom (GIS) score under treatment. Additionally, patients underwent a 13C octanoic acid breath test for the assessment of the gastric half‐emptying time (t1/2). Patients with prolonged t1/2 were diagnosed with gastroparesis and requested to repeat the test at the end of treatment. A change of t1/2 was defined a secondary study end point. t1/2 was prolonged in 48.6% of patients in the STW 5 group and in 43.8% of the placebo group. During treatment, t1/2 increased non‐significantly in patients treated with STW 5 (+23 ± 109 min; P = 0.51) and slightly accelerated among patients in the placebo arm (−26 ± 51 min; P = 0.77) (P = 0.49). The improvement of the GIS (P = 0.08) and the proportion of patients with a treatment response (P = 0.03) were more pronounced in the STW 5 group. Our findings suggest that the clinical effects of STW 5 in patients with FD and gastroparesis are not directly mediated by an acceleration of gastric emptying. A clear‐cut correlation with symptom improvement is still lacking.
Crossed Immunoelectrophoresis
Authors: Weeke Bent
Published: February 2009

Synonym— Laurell antigen‐antibody crossed electrophoresis (82) ad modum Clarke & Freeman (29).
13.α1‐Fetoprotein
Authors: Nørgaard‐Pedersen Bent
Published: February 2009

Summary. Rocket immunoelectrophoresis suitable for both quantitation and qualitative identification of α1‐fetoprotein is presented. By a slight modification of the rocket immunoelectrophoresis it is possible to detect α1‐fetoprotein in serum in a concentration of about 0.1 mg/1. A simple method for the final purification of α1‐fetoprotein is described. The concentration of α1‐fetoprotein in cord blood from 258 newborn infants is shown to be closely related to the gestational age of the newborn infants. The confidence limits for the estimation of gestational age are± 16 days.
15. Non‐Plasma Proteins in Cerebro‐spinal Fluid
Authors: BOCK ELISABETH
Published: February 2009

Summary. The tissue specificity of non‐plasma cerebrospinal fluid antigens was studied by various quantitative immunoelectrophoretic techniques using antisera against cerebrospinal fluid, brain, serum, urine, liver, erythrocyte membrane, immunoglobulins G, A, and M, λ and χ chains, fibrinogen, plasminogen, α2microglobulin, and β2microglobulin. Furthermore, CSF antigens were compared to antigens of plasma, urine, and extracts of brain, liver, muscle, and erythrocytes. The antiserum against brain was able to precipitate 11 antigens in the CSF. Two of these antigens were only found in CSF and brain extract.
19. Detergent‐Containing Gels for Immunological Studies of Solubilized Erythrocyte Membrane Components
Authors: BJERRUM OLE J. , LUNDAHL PER
Published: February 2009

Summary. Human erythrocyte membranes (ghosts) were dissolved to an extent of about 90% by repeated treatments at — 5° with the non‐ionic detergent Berol EMU‐043 in a concentration of 0.5% (w/v), and at pH 8.7. The solubilized membrane components were subjected to crossed immunoelectrophoresis with rabbit antibodies against ghost material. With 1% of Berol EMU‐043 in the gels, a recognizable pattern of 12 precipitation lines appeared. Only one of these was caused by a serum protein (albumin). The pattern of precipitates varied somewhat, probably due to some degradation of antigens by membrane‐bound proteolytic enzymes. Without detergent in the gels, a similar precipitation pattern was obtained, but some of the precipitation lines differed in shape and in staining intensity. In model experiments with human serum proteins, the incorporation of various non‐ionic detergents (Berol EMU‐043, Tween 20 SD, Triton X‐100, Lubrol WX, Nonidet P‐40) in a concentration of 2% (w/v) or less in the gels did not prevent immunoprecipitation of any protein.
22. Reversed Rocket Immunoelectrphoresis
Authors: AXELSEN NILS H. , SVENDSEN P. JUST
Published: February 2009

Summary. Rabbit and horse anti‐albumin were at pH 4.85 forced through albumin‐containing agarose gel, the result being well‐defined rocket‐shaped precipitates. The heights of these “reversed” rockets were linearly correlated to the antiserum concentration, and the method was therefore well suited for titre determination in large series of antibody samples.
Synaptic Plasma Mèmbrane Antigen D2 Measured in Human Cerebrospinal Fluid by Rocket‐Line Immunoelectrophoresis. Determination in Psychiatric and Neurological Patients
Authors: JøRGENSEN O. S. , BOCK ELISABETH
Published: February 2009

The concentration of an antigen partially identical to the rat brain synaptic membrane antigen D2, was measured in human cerebrospinal fluid. The method used was rocket‐line immunoelectrophoresis in agarose gel containing the detergent Triton X‐100, and line‐forming extract of rat brain membranes. No significant differences were found between the controls and patients with multiple sclerosis, inflammatory‐degenerative neurological diseases, atrophic brain diseases, acute cerebro‐vascular disease, schizophrenia, or affective psychosis.
Demonstration of Enolase Activity Connected to the Brain‐Specific Protein 14‐3‐2
Authors: BOCK ELISABETH , DISSING J.
Published: February 2009

An enzyme activity, enolase (2‐phospho‐D‐glycerate hydro‐lyase, E.C. 4.2.1.11), was shown to be connected to the brain‐specific protein 14.3.2. Specific staining methods were used on immuno precipitates of the protein. The enzyme activity was strongly inhibited by the addition of fluoride in the presence of phosphate. Metal ion binding capacity of the protein was shown by means of an auto radiographic method.
Demonstration of Circulating IgG‐Lactate Dehydrogenase Immune Complexes by Crossed Immunoelectrophoresis
Authors: GRUBB A. O.
Published: February 2009

Modified crossed immunoelectrophoresis was used to demonstrate complexes between IgG and lactate dehydrogenase in a human serum. The complexes were probably antigen‐antibody complexes, since the Fab fragments, but not the Fc fragments, of the IgG molecules combined with normal human lactate dehydrogenase. The IgG autoantibodies reacted with both types of poly‐peptide chains present in lactate dehydrogenase. The antibodies did not lower the catalytic activity of any of the 5 isoenzymes normally occurring in human serum. The antibodies formed exclusively soluble complexes with the isoenzymes and reacted not only with human lactate dehydrogenase, but also with that of rabbit and cow.
Detection of Membrane‐Bound Complement Components by Crossed Immunoelectrophoresis of Target Membranes Solubilized with Non‐Ionic Detergent
Authors: BHAKDI S. , BJERRUM O. J. , KNUFERMANN H.
Published: February 2009

Sheep erythrocyte membranes lysed with human complement were solubilized in 1% Berol EMU‐043 (w/v) and subjected to crossed immunoelectrophoresis in the presence of this detergent. Using antibodies to human serum proteins, we found 5–6 protein precipitates whose presence was dependent on complement action. Among these, C3 and C4 components of complement were identified. Membrane‐bound C5 (C5b) did not precipitate in crossed immunoelectrophoresis, but its presence on target membranes was demonstrated by crossed‐line immunoelectrophoresis (in situ absorption) of specific anti‐CS antibodies. A reaction of partial identity between membrane‐bound C5 (CSb) and serum C5 was found. Our experimental approach allows immunoelectrophoretic analysis of complement components at the membrane level and should therefore prove useful for future investigations on the molecular structure of membrane‐bound complement.
Immunoelectrophoretic Analysis of Sodium Dodecylsulphate‐Treated Proteins
Authors: NIELSEN C. SCHäFER , BJERRUM O. J.
Published: February 2009

Denaturation, following treatment with sodium dodecylsulphate, of human serum proteins and of membrane proteins from bovine milk fat globules was studied by means of quantitative immunoelectrophoretic techniques. As little as 1–2 mM of free sodium dodecylsulphate in a 5 μl sample was found to disturb the normal precipitation patterns. Furthermore the treatment resulted in the absence of certain precipitates, while other precipitates showed several peaks characteristic of proteins having partial identity. Considerable 'normalization' of the precipitation pattern was observed when non‐ionic detergent (Berol EMU‐043 and Triton X‐100) was incorporated in the samples or the gel prior to immunoelectrophoresis. Thus, this procedure renders the immunochemical analysis of proteins solubilized and/or fractionated in the presence of sodium dodecylsulphate more reliable.
Fractionation of PHA by Crossed Immunoelectrophoresis and Specific Immunosorption
Authors: KOCH C. , NIELSEN H. E.
Published: February 2009

Monospecific antibodies to 4 individual immunogenic components of a commercial PHA preparation were raised by immunization with the precipitates from crossed immunoelectrophoresis. The antibodies were used to prepare immunosorbents, which were then employed for the purification of individual PHA components. The results show that all the mitogenic and haemag‐glutinating activity of the crude PHA resides in one of the 4 immunogenic components.
Interactions between Ricinus Agglutinin and Human Plasma Proteins
Authors: HARBOE M. , SALTVEDT E. , CLOSS O. , OLSNES S.
Published: February 2009

Ricinus agglutinin purified to homogeneity precipitates certain serum glyco‐proteins containing terminal non‐reducing galactose residues. Immunoelec‐trophoresis of human sera in agarose gel with ricinus agglutinin in the antibody trough gave 2 fusing precipitin lines due to reaction with IgM and haptoglobin. All of 50 monoclonal IgM proteins precipitated with ricinus agglutinin, and the reactive sites were localized to the Fc μ fragment. By affinity chromatography on Sepharose columns containing insolubilized ricinus agglutinin prealbumin, albumin, α1‐lipoprotein, α1‐antitrypsin, α1‐acid glycoprotein, α1‐antichymotrypsin, α2HS glycoprotein, Ge‐globulin, caerulo‐plasmin, β‐lipoprotein, β1 C‐globulin, and transferrin were not retained, whereas α2‐macroglobulin, haptoglobin, IgM, IgA, and IgG were retained and could be eluted with lactose. Fused rocket immunoelectrophoresis revealed varying degrees of microheterogeneity among the latter proteins; almost all of IgM but only a small fraction of polyclonal IgG was retained on the ricinus column.
Identification of Glycoproteins with One and with Two or More Binding Sites to Con A by Crossed Immuno‐Affinoelectrophoresis
Authors: BøG‐HANSEN T. C. , BROGREN C.‐H.
Published: February 2009

Glycoproteins with one binding site and glycoproteins with two or more binding sites to concanavalin A may be identified by crossed immuno‐affinoelectrophoresis with free. and with immobilized concanavalin A. Human urinary acid phosphatase and human serum cholinesterase were demonstrated to be molecules with one and two or more binding sites to concanavalin A, respectively.
Characterization of Bovine Whey Proteins by Crossed Immunoelectrophoresis
Authors: LøWENSTEIN H. , BJERRUM O. J. , WEEKE EVA , WEEKE B.
Published: February 2009

A reference pattern obtained by crossed immunoelectrophoresis of bovine whey proteins from mature milk, using rabbit antibovine whey antibodies, has been worked out. Out of the total of 29 easily recognizable precipitates, 5 milk‐specific proteins and 21 serum proteins were identified. Furthermore, whey proteins which also occur in bovine milk fat globule membrane preparations and in extracts of bovine hair and dandruff were identified. The use of the obtained reference pattern for the identification of milk allergens and the determination of the specificity of human milk precipitins against bovine whey proteins is outlined.
Cross‐Reactions between Mycobacteria
Authors: CLOSS O. , HARBOE M. , WASSUM ANNE M.
Published: February 2009

Crossed immunoelectrophoresis (CIE) has been used to characterize the soluble antigens of Mycobacterium lepraemurium (Douglas strain) (MLM). The antigen was prepared by sonification of intact bacilli, purified as described. A pool of immunoglobulin isolated from hyperimmune rabbit antisera was used as antibody. By this procedure 42 antigens from MLM could be defined in a highly reproducible manner. An analysis of crossed line immunoelectrophoresis (CLIE) and CIE with intermediate gel as methods for studying antigenic cross‐reactivity are presented. Based on this analysis both BCG antigen and BCG antibody were added separately to the intermediate gel of the MLM reference system to study the cross‐reactivity between this mycobacterium and MLM. It was concluded that 23 out of the 42 MLM antigens defined in our reference system showed various degrees of cross‐reactivity. However, most of these cross‐reactions were weak. The method appears to be well suited for studying cross‐reactions between mycobacteria, but similar studies using other mycobacteria are needed before it can be decided how closely MLM is related to BCG.
Production of Antiserum against a Specific Herpes Simplex Virus Type 2 Antigen
Authors: VESTERGAARD B. F.
Published: February 2009

Antisera were produced in rabbits by inoculation of precipitates cut out of second dimensional agarose gels from crossed immunoelectrophoreses. The precipitating antigen chosen (antigen no. 1) had previously been identified as specific for herpes simplex virus type 2. Crossed immunoelectrophoretic analysis of the produced antisera revealed that the antisera contained a high concentration of precipitating antibodies that reacted exclusively with antigen no. 1 and a minor antigen with the same electrophoretic mobility. The produced antisera were useful in the typing of herpes simplex virus isolates by rocket immunoelectrophoresis.
Crossed Immunoelectrophoretic Analysis of Precipitins to Candida albicans in Deep Candida Infection
Authors: AXELSEN N. H. , BUCKLEY HELEN R. , DROUHTE E. , BUDTZ‐JøRGENSEN E. , HATTEL T. , ANDERSEN P. LEHM
Published: February 2009

By means of crossed immunoelectrophoresis with intermediate gel the precipitins to C. albicans in sera from 18 patients with deep Candida infection were compared with the findings in a control material consisting of sera from 169 'normal' controls (mainly healthy blood donors), 25 patients with oral candidiasis, 14 patients with vaginal candidiasis, and 6 persons without candidiasis giving false positive reactions by the double diffusion method. Up to 20 different precipitins were found in individual sera, and precipitins were demonstrated in all sera except in 6 'normal' controls. Of 692 precipitins 682 (98%) were identified and titrated in terms of the 78‐component reference system described earlier for C. albicans. No free Candida antigens were found, but would have been detected by the method. The best criterion for discrimination between sera from patients and controls was the precipitin score, which accounts for number of precipitins and for titres. A precipitin score of more than 6 1/2 had a diagnostic sensitivity of 88.9% and a specificity of 99.5%. Eleven precipitin specificities were found solely in the patients' sera, but the highest sensitivity for a single precipitin was 33%. Until more evidence has accumulated in favour of a monospecific immunoassay it is therefore rational to use a complex antigen extract for further evaluations of diagnostic Candida serology and a short cut to standardization would be the introduction of the method used in the present study.
Fractionation of Immunocompetent Cells with Different Physical Properties
Authors: HÄYRY PEKKA , ANDERSSON LEIF C.
Published: February 2009

Abstract. This paper deals with recent developments in the fractionation of immunocompetent cells with different physical properties, using preparative free flow cell electrophoresis, fractionation of cells according to their size and density.
SRBC Rosette Formation as a Human T Lymphocyte Marker
Authors: JONDAL MIKAEL
Published: February 2009

Abstract. This paper describes a technique utilizing rosette formation with sheep red blood cells as a human T lymphocyte marker, and separation of T cells by rosette sedimentation.
Lymphocyte Binding of Aggregated Immunoglobulin
Authors: DICKLER HOWARD B.
Published: February 2009

Abstract. This paper describes lymphocyte binding of aggregated immunoglobulin, which appears to bind to B lymphocytes and the third lymphocyte‐like cell population. The applications of aggregated Ig binding are also discussed, and the relation to Fc receptors and Ia‐like antigens.
Virus ‘Receptors’ on Lymphocytes
Authors: GREAVES M. F.
Published: February 2009

Abstract. This paper gives a review of the field of virus‐lymphocyte interactions and virus receptors on lymphocytes as well as the use of viruses as markers for various lymphocyte populations.
The Concept of Endodermal Sinus (Yolk Sac) Tumour
Authors: TEILUM G.
Published: February 2009

The concept of origin and natural history of endodermal sinus tumours (yolk sac tumours) is reviewed. These tumours were recognized and defined on a comparative histogenetic‐embryologic basis as germ cell tumours with selective overgrowth of the extraembryonic endoderm and mesoblast of the yolk sac. The concept of endodermal sinus tumour has created solid and rational basis for the study of alpha‐fetoprotein reappearance in a formerly heterogeneous and confusing group of tumours, whether they occur in the adult or in infantile gonads, or in such extragonadal sites as the mediastinum, the sacrococcygeal region, the pineal gland, the retroperitoneum or the infant vagina. Alpha‐fetoprotein studies, in turn, have furnished new evidence in favour of the concept.
Histological Patterns in Germ‐cell Tumours associated with Raised Serum Alpha‐fetoprotein (AFP)
Authors: TALERMAN A. , HAIJE W. G. , BAGGERMAN L.
Published: February 2009

Serum alpha‐fetoprotein (AFP) was studied in 237 patients with germ cell neoplasms of the gonads and extra‐gonadal sites. Whenever possible serial determinations were performed. All patients in remission had normal serum AFP. Raised serum AFP was seen in patients with active disease whose tumours contained endodermal sinus tumour (EST). This included 29 patients with testicular tumours, 11 with ovarian, 3 with mediastinal, and one with a retroperitoneal tumour. In all these patients serum AFP levels were high and correlated well with the amount of EST within the tumour. Serum AFP became normal when the disease was eradicated. There was a gradual rise with recurrence of the disease. Slightly raised serum AFP (below 60 ng/ml) was present in two patients with embryonal carcinoma examined preoperatively. In four similar cases serum AFP was normal. All patients with pure teratoma had normal serum AFP. Seventeen patients with pure seminoma whose serum was examined preoperatively had normal serum AFP at all times. These findings indicate that raised serum AFP is associated with the presence of EST within the tumour, and AFP is a good and reliable tumour marker in patients with germ cell neoplasms containing EST.
The Dynamics of Serum Alpha‐fetoprotein in the Course of Testicular Teratoma
Authors: KOHN J.
Published: February 2009

A retrospective study was carried out on a total of 279 cases of teratoma, out of which 135 were AFP positive, and 123 cases of seminoma. The AFP patterns were established by serial serum AFP assays carried out by radioimmunoassay at appropriately spaced time intervals. The following patterns could be distinguished: (1) Serum AFP levels normal throughout the whole course of the disease; (2) AFP levels consistently elevated irrespective of treatment; (3) normal levels for varying periods of time, then rising consistently; (4) elevated levels on presentation falling to normal for varying periods of time; (5) fluctuating with treatment, occasionally dropping to normal levels. Sequential AFP assays in patients with teratoma following orchidectomy, without evidence of residual tumour, suggest a half‐life of circulating AFP of approximately five days. A study was made to correlate any subsequent recurrence or absence of AFP elevation with the actual level of the patient's T½, compared with that of a normal T½ of five days. Appropriate formulas can be used to determine the patient's serum AFP T½ or to calculate the expected AFP level assuming a standard T½ of five days. The results suggest that a prolonged T½ is usually associated with recurrent elevations of serum AFP.
Multiple Tissue Markers in Human Malignant Testicular Tumours
Authors: HEYDERMAN E.
Published: February 2009

Using an indirect immunoperoxidase technique, the localization of human chorionic gonadotrophin (HCG), human placental lactogen (HPL), carcinoembryonic antigen (CEA) and alphafoetoprotein (AFP) has been achieved in paraffin embedded sections of human malignant testicular tumours. HCG was localized in the syncytiotrophoblast of trophoblastic teratomas, in morphologically distinctive cells of many teratomas classified as ‘non‐trophoblastic’ and in some seminomas. The presence of such cells showed good correlation with the development of elevated serum/urine HCG levels on or before overt tumour recurrence. HPL had a similar but more restricted distribution. CEA was demonstrated in areas showing intestinal‐like differentiation. AFP could be readily demonstrated in tumours with yolk sac differentiation, although widespread diffusion made accurate localization difficult.

The routine localization of these tumour products may be of potential value in improving the diagnosis and classification of malignant testicular disease. Preliminary results also indicate that such an approach may have a role in predicting suitable serum markers with which to monitor disease activity and in providing further prognostic criteria.
α‐Fetoprotein Synthesis in Yolk Sac Tumor: Sex‐dependent Production of α‐Fetoprotein by Transplantable Rat Yolk Sac Tumor
Authors: ENDO Y. , KANEKO Y. , URANO Y. , TSUCHIDA Y. , WATABE H. , TSUKADA Y. , SAKASHITA S. , HIRAI H. , ODA T.
Published: February 2009

Transplantable rat yolk sac tumor cells produce α‐fetoprotein (AFP) and their ability to synthesize AFP is maintained for generations in female rats. However, when they are transplanted in male rats, the AFP production decreases markedly.

To examine this sex dependency in AFP production, experiments at cellular and subcellular levels were carried out. In cell incubation studies, yolk sac tumor cells maintained in female rat (YST‐F cells) synthesized more AFP than yolk sac tumor cells maintained in male rat (YST‐M cells). Using cytosol RNAs prepared from YST‐F and YST‐M cells, AFP production was studied in cell‐free protein synthesizing system derived from wheat germ.

In this system, cytosol RNA from both YST‐F and YST‐M cells directed AFP synthesis. But the amount of AFP synthesized was smaller in the presence of RNA from YST‐M cells. From these results, it was suggested that the reduced AFP synthesis by YST‐M cells is, at least partly, due to a quantitative decrease in their cytosol messenger RNA coded for AFP.
Mouse Alpha‐fetoprotein: Genetic Studies
Authors: LINDAHL G. , OLSSON M. , RUOSLAHTI E.
Published: February 2009

The genetic control of alpha‐fetoprotein (AFP) synthesis in the mouse is studied. Structural variants of mouse AFP have not yet been found, although 22 inbred strains have been tested for the presence of such variants. In constrast, a mouse strain has been found, in which the regulation of AFP synthesis is altered. In this mouse strain the rate at which AFP synthesis is turned off after birth is decreased, as compared to other inbred mouse strains. This property is controlled by a single recessive Mendelian gene, which has been called Raf (for regulation of alpha‐fetoprotein synthesis). The conclusion that Raf is a single recessive Mendelian gene is supported both by genetic data published previously and by new genetic data presented in this paper. The Raf gene not only controls the rate at which AFP synthesis is turned off after birth, but also controls the amount of AFP produced under conditions of liver regeneration. Thus, characterization of the Raf gene can shed light both on the mechanism that turns off AFP synthesis after birth and on the mechanisms that control regeneration of the liver.
A Sensitive and Simple Determination of Human α‐fetoprotein by Enzyme‐electroimmunodiffusion
Authors: NISHI SHINZO , FUJIYAMA YOSHIHIDE , HIRAI HIDEMATSU
Published: February 2009

A simple and sensitive determination method of α‐fetoprotein was developed by introducing α‐fetoprotein horseradish peroxidase conjugate into electroimmunodiffusion.

The precipitin line formed was directly visualized by staining for peroxidase activities. The sensitivity was 32 ng/ml. Electrophoretic activities of AFP were analyzed by two dimensional electrophoresis applying the present method to the second electrophoresis.

AFP from normal adults and several patients was confirmed to migrate as an α‐globulin as AFP of hepatoma, yolk sac tumor and fetuses did.

AFP‐like activities were detected in β‐γ‐globulin region in some samples but these were not due to AFP‐anti‐AFP interactions.
Alpha‐fetoprotein Determination: A Warning
Authors: MARRINK J.
Published: February 2009

Alpha‐1‐fetoprotein (AFP) was determined by different immunochemical techniques in the serum of a patient with liver cirrhosis. Although a precipitation line was observed against anti‐AFP antiserum, using bidimensional immunodiffusion, immunoelectrophoresis, countercurrent electrophoresis and rocket electrophoresis, this precipitation line however showed a reaction of non‐identity with a positive AFP control. Several different commercial anti‐AFP antisera gave the same false‐positive result. Analysis of the patient's serum revealed the falsely positive reacting component to be of IgM‐like nature, having a cathodal mobility. These findings are discussed in the light of the determination of slightly elevated AFP levels by a method such as the radioimmunoassay, not allowing identification by means of identity with control specimen. The consequence of our results is also discussed with respect to normal serum AFP.

The described non‐AFP activity was detected regularly in sera taken over the period 1969–1976.
Concanavalin A‐affinity Molecular Variants of Rat AFP: Differences Among Amniotic Fluid, Fetal Serum, Newborn Serum and Maternal Serum
Authors: SMITH C. J. , KELLEHER P. C.
Published: February 2009

The concanavalin A‐affinity molecular variants of rat alpha‐fetoprotein were measured in amniotic fluid, fetal serum, newborn serum and maternal serum. During the last 6 days of gestation, 54% ± 2% (mean ± S.E.M.) of the amniotic fluid alpha‐fetoprotein does not react with concanavalin A. At 17 days gestation, 54% ± 4% of the fetal serum alpha‐fetoprotein does not react with concanavalin A. Between 18 and 21 days gestation the percentage of the concanavalin A‐nonreactive molecular variant of fetal serum alpha‐fetoprotein decreases to 43% and remains at 43% ± 0.4% through the 28‐day newborn period studied. Following day 18 of gestation the proportions of the alpha‐fetoprotein concanavalin A‐affinity molecular variants in serum and in amniotic fluid are different (p<0.001). On day 21 of gestation, 41% of the alpha‐fetoprotein present in maternal serum does not react with concanavalin A. These results suggest that during late gestation yolk sac and fetal liver synthesize different patterns of alpha‐fetoprotein concanavalin A‐affinity molecular variants and that on day 21 of gestation the major portion of the maternal serum alpha‐fetoprotein is derived from fetal serum.
Purification and Physical Characterization of Human Alpha‐fetoprotein from Cord Blood and from Ascites Fluid of Individuals with Hepatocellular Carcinoma
Authors: KORTRIGHT K. H. , KAPADIA G. G. , LEE S. Y. , McINTIRE K. R. , WALDMANN T. A.
Published: February 2009

Human alpha‐fetoprotein (HuAFP) was isolated from cord blood and ascites fluid of individuals with hepatocellular carcinoma. The purification procedures employed antibody‐affinity chromatography and preparative polyacrylamide gel electrophoresis (Prep PAGE). Final yields of HuAFP averaged 40% and demonstrated 99.8% purity.

Physicochemical studies of HuAFP from cord blood showed a molecular weight of 70.000, pl = 4.85, Stokes radius of 2.699 nm, and valence of (H*/mol) of 11.0. Similar studies of HuAFP from ascites fluid from individuals with hepatocellular carcinoma showed a molecular weight of 69.000, pl = 4.80, Stokes radius of 2.798 nm. and valence of (H*/mol) of 12.2. These AFP molecules isolated from separate origins demonstrated no significant physicochemical or antigenic differences. Assessment of potency and purity of these preparations were made by Lowry protein analyses, radioimmunoassay (RIA) to AFP and albumin. PAGE, SDS‐PAGE, and isoelectric focusing patterns on polyacrylamide gel electrophoresis.

The sensitivity of the RIA for human AFP was increased by the use of high affinity goat anti‐AFP. To minimize non‐specific serum effects, human serum, from which AFP had been specifically removed by affinity chromatography was added to each tube in the standard dose response curve. The sensitivity of the assay was 0.7 ng/ml.