A new approach for evaluation of the in vitro haemolytic potential of a solution of a new medicine

In the process of developing an intravenously injectable drug, its haemolytic potential must be considered. There are no Regulatory Guidelines for this kind of test. Many authors have set up different models, attempting to obtain early information about the behaviour of test compounds when injected into the bloodstream.In the present work, an in vitro static model is presented, which takes into account the injection rate (R _inj.) of the drug, and the blood flow rate (Q _v) of the vein in which the drug must be injected. From the relationship between these two parameters, the C_max, expressed as mg/ml, can be calculated. This latter parameter allows us to calculate the drug concentration which, at any moment during injection, comes into contact with a known aliquot of new' blood passing through the injection site. Furthermore, a dynamic test has been developed, which simulates an injection into the blood flow using a tubing system and infusion pumps set for the same R _ini. and Q _v values used in static test. Two injectable drugs, Valium® and Lanoxin®, and a commonly used vehicle, propylene glycol, have been tested by both the methods. These compounds have also been tested with another in vitro method (Prieur et al. 1973), in which a volumetric blood-to-test solution ratio of 1:1 is adopted for every drug tested, with neither R _inj. nor Q _v being taken into account. Results of the haemolytic potential obtained with the three tests have been compared.A good correlation has been observed between the static and the dynamic tests, whereas Prieur's model, which uses a drug-to-blood ratio which is far higher than in vivo, has been shown to give false positive results.It is concluded that a test for the evaluation of the haemolytic potential of drugs must take into account the pharmacodynamic characteristics of the formulation intended to be injected, and at least the blood flow rate. The proposed static test has been demonstrated to be an easy and reliable method of obtaining a true picture of the in vivo situation.     

The oligodendroglial reaction to brain stab wounds: An immunohistochemical study

Summary Myelin/oligodendrocyte specific protein was compared to glial fibrillary acidic protein and 23-cyclic nucleotide 3-phosphodiesterase expression in normal rat brains and following stab wounds to the cerebral cortex, corpus callosum and hippocampus. Animals with stab wounds were allowed to recover for 5,15,28, 45 and 70 days post-operation before fixation by perfusion. Sections were reacted with antibodies against myelin/oligodendrocyte specific protein, glial fibrillary acidic protein and 23-cyclic nucleotide 3-phosphodiesterase, and observed by light and electron microscopy. Normal cerebral cortex had very few myelin/oligodendrocyte specific protein-positive and 23-cyclic nucleotide 3-phosphodiesterasepositive cells, but some glial fibrillary acidic protein-positive cells. The myelinated fibres of the corpus callosum were heavily stained for myelin/oligodendrocyte specific protein but unstained by glial fibrillary acidic protein or 23-cyclic nucleotide 3-phosphodiesterase antibodies. Some immunopositive cells were present in the corpus callosum and hippocampus with all three antibodies. After stab wound myelin/oligodendrocyte specific protein-positive reactive cells had more and longer processes and stained more intensely than equivalent cells in normal brain. These cells were distributed along the wound track, including within the cerebral cortex. The numbers of these cells increased until 28 days post-operation and then decreased so that very few were found at 70 days post-operation except in the corpus callosum. Where demyelination occurred myelin/oligodendrocyte specific protein-staining was lost. Staining for 23-cyclic nucleotide 3-phosphodiesterase revealed a similar pattern. Glial fibrillary acidic protein-positive reactive cells, which were also more robust than the normal cells, were more widely distributed. They increased in number throughout the time periods studied and gliosis was evident on the contralateral side. The glial fibrillary acidic protein-positive astrocytes were also different from the myelin/oligodendrocyte specific protein-positive and 23-cyclic nucleotide 3-phosphodiesterase-positive oligodendrocytes in terms of cell shape. With electron microscopy myelin/oligodendrocyte specific protein-positive cells showed features typical of immature oligodendrocytes. We conclude that the injury caused a numerical increase in oligodendrocytes and that myelin/ oligodendrocyte specific protein is a good marker for the oligodendroglial response and demyelination in pathological conditions.     

The influence of display characteristics on active pursuit and passively induced eye movements

Summary A series of experiments has been conducted on human subjects to examine the effect of the movement of small targets located in the peripheral visual field on oculomotor response. Subjects were presented with either a single centrally positioned target or a pair of targets displaced at angles of ±5°, ±10° and ±20° from centre. Target movement was in the horizontal plane, the paired targets always moving in unison. The stimulus waveform consisted of either a sinusoidal or random target motion encompassing a frequency range from 0.1 to 4 Hz with an angular displacement of ±3.5°. Subjects made two types of response. First they were instructed to follow the single target or the centre point of the paired targets. In this active pursuit condition the gain of slow-phase eye velocity progressively decreased as the moving targets were moved from the central position to the most peripheral location (±20°). Secondly, subjects were required passively to ignore the target movement by staring blankly ahead. During this passive response nystagmic eye movements were induced for which the slowphase eye velocity also decreased with increasing target eccentricity, but the gains were always less than those induced during active pursuit. The frequency characteristics of the passive response were very similar to those of the active response, breaking down at frequencies beyond 1 Hz. The ability to suppress the passive response was also investigated by the presentation of a tachistoscopically illuminated earth-fixed target. The response was found to decline as the interval between presentations of the fixation target was decreased from 3000 ms to 100 ms. It is suggested that the passive response originates from a basic velocity drive to the oculomotor system resulting from image movement across the retina. This velocity drive may be cancelled with adequate fixation but must be enhanced to accomplish desired eye velocity during active pursuit.     

alpha1-acid glycoprotein possesses in vitro pro- and antiinflammatory activities

We studied the effects of ₁-acid glycoprotein on tumor necrosis factor- (TNF-) and interleukin-10 (IL-10) production and lymphocyte response to phytohemagglutinin in cultured peripheral blood mononuclear leukocytes from 6 healthy donors. We observed 2 opposite responses to ₁-acid glycoprotein: first, stimulation of TNF- and IL-10 production and inhibition of lymphocyte proliferation, and second, suppression of cytokine production and stimulation of lymphocyte proliferation. In cell cultures isolated from 4 of 6 donors, the TNF-/IL-10 ratio remained unchanged after addition of native ₁-acid glycoprotein, but some fractions isolated by chromatography on concanavalin A-Sepharose changed this parameter. These changes were most pronounced after treatment with fraction C enriched with molecules with incomplete (biantennary) carbohydrate chains. The mechanisms of ₁-acid glycoprotein-induced effects on peripheral blood mononuclear leukocytes are discussed.     

Recombination of yeast mitochondrial DNA does not require mitochondrial protein synthesis

Summary Mitochondrial genes recombine extensively in yeast zygotes. In heteropolar crosses (⁺ × ^–) in which the ^– allele consists of an insertion, there is preferential recovery of ⁺ and markers closely linked to it. This polarity has been postulated to be a consequence of one-way gene conversion beginning at the locus (- to ⁺). We have shown that most or all mitochondrial recombination in homopolar and heteropolar crosses, and the phenomenon of polarity itself, does not require products of protein synthesis on mitochondrial ribosomes. (i) Yeast strains were grown and mated, and the zygotes plated and grown, on glucose medium with erythromycin to inhibit and dilute out the products of mitochondrial protein synthesis. Recombination frequencies and polarity at the cap1 and oli1 loci were normal compared to controls in some homopolar (⁺ × ^–) and heteropolar crosses. Apparent changes in recombination frequencies and polarity were seen in other crosses but are attributable to locus-specific petite induction by erythromycin. (ii) Homopolar (⁺ × ⁺) and heteropolar crosses between pairs of petite mutants retaining the cap1, ery1, and oli1 loci also showed nearly normal recombination at the cap1 and oli1 loci, as determined by test-crossing the petite progeny. The petite mutants and zygotes cannot do mitochondria) protein synthesis. These results support the recombinational model of polarity.     

Expression of parathyroid hormone receptors in MDCK and LLC-PK1 cells

Parathyroid hormone (PTH) inhibits renal proximal tubular phosphate (P_i) and bicarbonate reabsorption by regulating the activity of apical Na/P_i cotransport and Na/H exchange. Two renal epithelial cell lines [proximal tubular, LLC-PK₁; distal tubular, Madin-Darby canine kidney, (MDCK) cells] were stably transfected with complementary deoxyribonucleic acids (cDNAs) encoding a cloned PTH receptor in order to examine the polarity of transfected receptor function and whether or not intrinsic P_i transport is regulated by the transfected PTH receptor. The receptors are functionally coupled to the stimulation of adenosine 35 cyclic monophosphate (cAMP) production at both cell surfaces in LLC-PK₁ cells, whereas this response is primarily limited to the basolateral surface in MDCK cells. Immunocytochemistry suggests an apical and basolateral localization of the transfected PTH receptor in LLC-PK₁ cells and only a basolateral localization in MDCK cells. PTH activation of the transfected receptors is not coupled to the regulation of intrinsic P_i transport in either LLC-PK₁ or MDCK cells.     

Chromaticity of synaptic inputs to H1 horizontal cells in carp retina: analysis by voltage-clamp and spectral adaptation

Summary Cone photoreceptor inputs to H1 horizontal cells (H1 HCs) in carp retina were studied by measuring light-modulated currents (I_L) to monochromatic stimuli (460, 533, 688 nm) under a voltage-clamp condition. By using double-barrelled micro-electrodes H1 HCs were voltage-clamped whilst perfusing with dopamine to uncouple the cells. The I_L of the H1 HCs driven by each cone input was segregated by selective chromatic adaptation, and differences in the kinetics of the I_L of the H1 HCs were revealed. Thus, all together, three types of I_L were observed: (1) a fast outward current to the long-wavelength stimulus; (2) a slow outward current to the middle-wavelength stimulus; and (3) a delayed inward current that followed the peak of slow outward current to the short-wavelength stimulus. The reversal potentials of the three currents were estimated to be at least 20 mV more positive than the dark resting potential by extrapolation of the I_L-V curve. These observations are consistent with the idea that the H1 HCs receive sign-inverting, conductance decreasing synaptic input(s) from at least one other cone mechanism, in addition to the main conventional EPSP type synaptic input from red-sensitive cones.     

Rhinovirus detection using probes from the 5′ and 3′ end of the genome

Summary This study investigated the abilities of cDNA probes from the 5 and 3 ends of the genome of human rhinoviruses (HRV-) 14, 9, and 1B to detect RNA from 59 rhinovirus serotypes. The results show that probes from the 5 end of the genomes of HRV-14, 9, and 1B detected a large number of serotypes but the detection rate was variable and depended on the degree of homology with the particular probe. In contrast, all the 3 end probes were specific for the homologous virus. However, along HRV-9 probe detected a large number of serotypes.It was concluded that such cDNA probes would not detect all serotypes with equal efficiency. Synthetic oligonucleotides corresponding to short but highly conserved regions in the 5 non coding region may overcome this problem.     

Viewing-distance invariance of movement detection

Summary Since visual movement information is often presented in electronic displays or films it is amazing that there is a paucity of research on the influence of viewing distance on motion detection in cinematograms. We report a relatively high degree of detection constancy with changing viewing distance for coherent motion in random-pixel cinematograms. A constant performance irrespective of viewing-distance is called distance-invariance and for motion detection it proves to hold reasonably well for a relatively wide range of viewing distances both for foveal and eccentric vision. The limits of this viewing-distance invariance are explored as a function of screen velocity. Detection performance is quantified by a theshold signal-to-noise-ratio (SNR-) value, S, which is determined as a function of velocity for a range of viewing distances from 53 to 13476 mm for foveal vision and from 60 to 1925 mm at 24° eccentricity on the nasal horizontal meridian of the right eye's retina. The data can be explained, at least qualitatively, by a model in which a spatial-resolution stack has a stack of velocity-tuned motion detectors at every resolution layer. Such a stack-of-stacks model is in line with proposals for contrast-detection stack-models, but it suggests that the usual hypothesis that motion perception is based on the activity of two separate systems, the short-range and the long-range system, might be superfluous. This two-systems distinction was largely based on the different performance found for moving random dot patterns and moving form-defined stimuli. A moving random pixel array viewed at very close range (e.g. 6 cm) presents the subject with relatively large almost square blobs, which are less dissimilar from the phi-stimuli used in classic motion perception studies than random dot stimuli at the usual medium to large viewing distances. It leads to maximum displacement threshold (D_m-) values that are not untypical of the long-range system, but by gradually increasing the viewing-distance and thus decreasing the pixel-size a continuous change is found from typical long-range to typical short-range values of D_m. The two-systems distinction for motion detection appears to refer to the stimulus rather than to the visual system: The motion-detection system might be forced into a local or a global mode of operation by the choice of stimulus.     

Zur Permeation der Serumeiweiße in die Haut und ihrer quantitativen Bestimmung am Modell der Cantharidenblase

Zusammenfassung Mittels chemischer und immunochemischer Methoden wurde vergleichend der Gehalt an Proteinen in Cantharidenblasen und Blutserum bestimmt. Erstmals werden auch quantitative Untersuchungen von Präalbumin, ₂-M-Globulin, Coeruloplasmin, Haptoglobin und Transferrin im Cantharidenblaseninhalt vorgenommen. Dabei fanden sich statistisch gesicherte Unterschiede zwischen Cantharidenblasen- und Seruminhalt im Sinne tieferer Werte in den Blasen bei Gesamteiweiß, Präalbumin abs., ₂-M-Globulin abs., ₂-Globulin abs. und rel.-%, Coeruloplasmin abs., -Globulin abs., Transferrin abs. und -Globulin abs. Albumin abs. und rel.-% wiesen signifikante Unterschiede in Richtung höherer Werte in den Cantharidenblasen auf. Das Makroglobulin ₂-M-Globulin passierte vergleichsweise schwerer die Capillarschranke zwischen Blut und Blaseninhalt als die niedermolekularen Proteine Coeruloplasmin, Haptoglobin und Präalbumin.

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Summary Comparative studies on Protein contents of blood and Cantharidin blisters were performed by means of chemical and immunochemical methods. For the first time quantitative measurements of Praealbumin, ₂-M-Globulin, Coeruloplasmin and Transferrin were done. Statistically significant differences between blister content and serum levels of proteins with higher values in the blisters were found in Total Protein, Praealbumin abs., ₂-M-Globulin abs., ₂-Globulin abs and rel.-%, Coeruloplasmin abs., -Globulin abs., Transferrin abs and -Globulin abs. Only Albumin abs. and rel.-% showed higher values in blisters as compared with the serum levels. Praealbumin, Haptoglobin and Coeruloplasmin were found to pass the capillary membrane, as measured by the difference between serum and blisters fluid content, more easily than the macromolecular protein ₂-M-Globulin.

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Herrn Prof. Dr.H. G. Bode, Direktor der Univ.-Hautklinik Göttingen, zum 60. Geburtstag am 31.8.64 verehrungsvoll gewidmet.     

Regulated overproduction of α-amylase by transformation of the amylolytic yeast Schwanniomyces occidentalis

Summary High frequency transformation of a Schwanniomyces occidentalis mutant defective in the last step of tryptophan synthesis was achieved with plasmids containing the tryptophan synthetase gene (TRP5) of Saccharomyces cerevisiae and an autonomous replication sequence from S. occidentalis, which we called SwARS1. The SwARS1 fragment is also functional in S. cerevisiae. The average copy number of the plasmids in both yeast species was 5–10 per cell under selective conditions. S. occidentalis cells that were transformed with an autonomously replicating plasmid carrying the cloned -amylase gene from S. occidentalis secreted about five times more -amylase than cells without additional copies of the -amylase gene. Both the chromosomal copy and the plasmid-carried copies of the -amylase gene were repressed in the presence of glucose. This transformation system provides a possibility to improve starch degradation by S. occidentalis.     

Untersuchungen über den Fett-Transport im Serum mit der Lipidelektrophorese auf Membranfolie

Zusammenfassung An einem kasuistischen Material von 52 Fällen mit insgesamt 201 Untersuchungen wurde der Mechanismus des Fett-Transportes nach oraler und parenteraler Fettbelastung untersucht. Die Bestimmung der Lipoproteide erfolgte mit der Lipidelektrophorese auf Membranfolien.Die Transportfraktionen für die leichten Lipoproteide (Chylomikronen) wurden durch einen Vergleich des Lipidpherogramms vor und nach Eliminierung leichter Lipoproteide ermittelt. Wie die vorgelegten Resultate zeigen, wird das Neutralfett in der ₂-Lipoproteidfraktion transportiert, während die übrigen Fraktionen nur in geringem Umfang zu Transportzwecken verwendet werden. Oral oder intravenös zugeführtes Fett verhält sich gleich. Normalerweise kommt es nur zu einem vorübergehenden, kurzfristigen Anstieg der ₂-Lipoproteidfraktion. Bei gestörtem Fett-Transport finden sich bereits im Nüchternserum erhöhte Werte für die ₂-Lipoproteidfraktion, die auch nach Entfernen leichter Lipoproteide über die Norm vermehrt anzutreffen sind. Nach Fett-Belastungen kommt es in dieser Fraktion zu einem weiteren Zuwachs von Lipoproteiden. Der Anstieg hält länger an als gewöhnlich. Die Lipidelektrophorese auf Membranfolien erlaubt ferner eine Differenzierung vorwiegend hyperlipämischer und vorwiegend hypercholesterinämischer Seren. Eine Neutralfetthyperlipämie ist durch eine hohe ₂-Lipoproteidfraktion charakterisiert, während eine Hypercholesterinämie mit einer Zunahme der -Fraktion einhergeht.Die mit immunologischen Methoden festgestellten Veränderungen des Lipoproteidmusters bei gestörtem Fett-Transport lassen sich auch mit Hilfe der Lipidelektrophorese auf Folien nachweisen. Bei Fett-Transportstörungen nimmt die Wanderungsgeschwindigkeit der _1–A-und _1–B-sowie der -Lipoproteidfraktion zu.Die Vorteile dieser Technik gegenüber den bisher verwendeten Methoden werden diskutiert und die Bedeutung der Lipidelektrophorese auf Membranfolie für die Beurteilung der Serumlipoproteide und von Fett-Transportstörungen erörtert.

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Summary The mechanism of fat-transport after oral and parenteral fat-application has been explored on a choice of 52 cases with 201 findings all together. The analysis of lipoproteids was carried out with the membrane-lipid-electrophoresis. The vehicles for the light lipoproteids (chylomicrones) were stated by comparison of the lipid-pherograms before and after elimination of light lipoproteids.In the course of investigation we found, that the neutral fat is transported in the ₂-lipoproteid-fraction, whereas the other fractions are used for transport purposes only in a small extent. There is no difference in the transport-mechanism of fat, given orally or intravenously. Normal fat-transport is characterized by a transitory increase of the ₂-lipoproteidfraction. Irregular fat-transport shows a fat-increase within the ₂-lipoproteid-fraction in the serum of fasting patients. This fraction is to be found increased abnormally after having removes the light lipoproteids. After fat-application a further increase of lipoproteids in this fraction is to be noted. This increase lasts longer than usually.With membrane-electrophoresis it is possible to distinguish between the serum of the hyperlipemic and hypercholesterinemic type. Neutral fat-hyperlipemia is characterized by a high ₂-lipoproteid-fraction, whereas a hypercholesteremia is combined with an increase of the -fraction.Results, obtained by immunological methods concerning the lipoproteid pattern of irregular fat-transport could be confirmed with aid of the membrane-electrophoresis. Irregular fat-transport shows an increased speed of the _1–A and _1–B as well as the -lipoproteid-fraction on the acetate membrane.The advantages of this technique are discussed in comparison to methods hitherto practised and the importance of the membrane-electrophoresis is mentioned in regard to the judgement of the serum-lipoproteids and the irregular fat-transport.

     

Anti-CD3-induced changes in protein kinase C isozymes expression in human CD4+ and CD8+ T lymphocytes

In order to determine whether there is a differential expression and activation of PKC isozymes between CD4+ and CD8+ T cells, peripheral blood mononuclear cells were stimulated with anti-CD3 monoclonal antibody (moAb) for various time intervals and the expression of calcium-dependent PKC isozymes (, , ) and calcium-independent PKC isozymes (, , ) was analyzed with dual color flow cytometry, using anti-PKC isozyme antibodies and anti-CD4 or anti-CD8 antibodies. The basal fluorescence intensity of all PKC isozymes was comparable between CD4+ T cells and CD8+ T cells. Following activation with anti-CD3 moAb a marked increase in the fluorescence intensity of all PKC isozymes in both CD4+ and CD8+ T cells, albeit to a different extent and with different kinetics was observed. Among all PKC isozymes studied, the least striking changes were observed in PKC isozyme and the most striking changes were observed in PKC- isozyme. Laser-based confocal microscopic studies confirmed that the increase in fluorescence intensity of PKC isozymes following anti-CD3 moAb stimulation, as measured by flow cytometry was accompanied by the translocation of PKC isozymes from cytosol to the plasma membrane. This study demonstrates a differential effect of anti-CD3 moAb on the expression of PKC isozymes between CD4+ and CD8+ T cells and suggests that flow cytometry can be used to study the translocation of PKC isozymes from cytosol to the plasma membrane.     

An analysis of adrenergic influences on the sural-gastrocnemius reflex of the decerebrated rabbit

Summary The sural-gastrocnemius reflex was observed in decerebrated rabbits during intrathecal application of four -adrenoceptor antagonists. Idazoxan and yohimbine, which are antagonists at the ₂-receptor, caused facilitation of the reflex, although idazoxan was more potent and produced a larger overall increase in the reflex response. However, when given after yohimbine, idazoxan elicited no further increase in reflex responses. The differences between the two drugs may result from the interaction of yohimbine with receptors for 5-hydroxytryptamine. The selective ₁-receptor antagonist prazosin had no consistent effects when given alone, but reduced the facilitatory effects of idazoxan. The putative selective post-junctional ₂-receptor blocker SK&F 104078 had no significant effects when given alone, nor did it influence the facilitatory action of a subsequent dose of idazoxan. Section of the spinal cord in the presence of idazoxan always caused a decrease in gastrocnemius responses to sural nerve stimulation. These data show that the facilitatory effects of idazoxan are almost certainly mediated at the spinal cord and that they do not involve blockade of ₁-receptors. It appears that idazoxan acts by blockade of adrenergic descending inhibition in combination with increased descending facilitation. The inhibition is probably mediated through noradrenaline acting at ₂-receptors, and the facilitation may be the result of release of noradrenaline (acting at ₁-receptors) and 5-hydroxytryptamme in the spinal cord.     

Presumptive seventh serotype of human rotavirus

Summary Four group A human rotaviruses having antigenic specificity of subgroup I and long RNA electropherotype were isolated in MA104 cell cultures. Cross-neutralization tests with hyperimmune antisera suggested that they are serologically distinct from the six previously recognized human rotavirus serotypes.     

Effects of antisense oligodeoxynucleotide hybridization on in vitro translation of potato virus Y RNA

Potato virus Y (PVY), a potyvirus, has an RNA genome containing 9704 nucleotides of which 185 belong to the 5 nontranslated region (NTR). Contrary to most eukaryotic mRNAs that have a cap structure, the potyvirus RNA has a genome-linked protein (VPg). In order to understand the mechanisms of PVY RNA translation initiation, hybrid-arrest translation was used to localize sequences involved in binding of proteins and/or ribosomes. The 5 NTR was fused to the -glucuronidase (GUS) reporter gene. Six antisense oligodeoxynucleotides were used for hybridization, and the efficiency of the in vitro translation of the hybridized mRNA was modified to different levels depending upon the position of the oligodeoxynucleotide used. The highest inhibition was obtained with an oligodeoxynucleotide hybridized to the 5 end. In addition, translation of GUS mRNA containing the PVY 5 NTR was greatly enhanced when this mRNA was capped. These results differ from those obtained with the tobacco etch virus (TEV) and three picornaviruses, but are similar to those obtained with capped mRNA.     

Role of phosphatidylinositol-3-kinase and protein kinase Cζ in the adenylate cyclase signal mechanism of action of relaxin in muscle tissues of rats and mollusks

We showed that phosphatidylinositol-3-kinase and protein kinase C are involved in the adenylate cyclase signal mechanism of relaxin action. A selective inhibitor of phosphatidylinositol-3-kinase wortmannin blocked the stimulatory effect of relaxin on adenylate cyclase in rat skeletal muscles and Anodonta cygnea smooth muscles. Antibodies against protein kinase C abolished the relaxin-induced stimulation of adenylate cyclase in rat muscles, but not in mollusk muscles. Our results indicate that phosphatidylinositol-3-kinase and protein kinase C play a role in the adenylate cyclase signal mechanism of relaxin action.Translated from Byulleten Eksperimentalnoi Biologii i Meditsiny, Vol. 138, No. 10, pp. 420–423, October, 2004     

Immunochemical identification of organ-specific human placentalαin2-globulin and its content in the amniotic fluid

An organ-specific human placental ₂, differing immunologically from previously known ₂ and ₁-globulins of pregnancy, -fetoprotein, placental lactogen, and chorionic gonadotropin, was identified by immunochemical analysis. The antigen thus discovered is present in large quantities in the placental tissue and amniotic fluid in the early stages of pregnancy, but at parturition its concentration is sharply reduced.Presented by Academician of the Academy of Medical Sciences of the USSR Yu. M. Lopukhin.Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 82, No. 7, pp. 803–804, July, 1976.     

In vivo andin vitro tests in anaphylactic reactions to anaesthetic agents

The majority of patients with anaphylactoid reactions (AR) under general anaesthesia showed histamine release from basophil leucocytes (HRL) by neuromuscular blockersin vitro (none in normals) and high levels of serum IgE antibody (detected by the new paper radioallergosorbent test, RAST; little or none in normals) which reacted with the quaternary ammonium group of choline, and less frequently of alcuronium, strongly suggesting that IgE antibody, HRL and AR are related. Skin tests on their own are of limited value.