Antigenic cross-reactivity among rodent brain tissues and stem cells. Previous studies have shown that antisera prepared in rabbits against mouse brain (RAMB) contains activity in vitro against the mouse bone marrow colony-forming unit (CFU) or hematopoietic stem cell. In the present study, in vitro treatment of mouse bone marrow with antisera prepared in rabbits against brain tissue from rats (BARB) or hamsters (RAHB) also reduced the CFU content of the mouse marrow. Prior absorption of the RAMB serum with fetal liver tissues from rats or hamsters as well as mice reduced the anti-CFU activity of the RAMB preparation. In addition, absorption of the RAMB preparation with brain tissue from any of the rodents reduced the activity of the antiserum for the mouse stem cell. It appears that the determinants previously shown to be shared by the brain tissue and stem cells of mice are cross-reactive with determinants present on the stem cells and brain tissue of rats and hamsters.


A heterophile system in human renal transplantation. V. Relationship of heterophile transplantation antigen and common antigen of Enterobacteriaceae. Heterophile transplantation antigen and common antigen of Enterobacteriaceae appear serologically to be separate specificities. However, both antigens are common to Enterobacteriaceae, rat erythrocytes, and some human kidneys. Both antigens are obtained from various tissues by the same chemical procedure. Immunity to each antigen is frequently produced by renal transplantation. We suggest that the antigens are either separate molecules which are similar in chemical structure in the region of the antigenic determinant as well as in tissue distribution or separate reactive sites located on the same molecule. The possibility that common antigen may be a human alloantigen raises theoretical possibilities relative to susceptibility to infection and pyelonephritis, as well as to its relationship to histocompatibility.


Hemagglutination inhibition studies for the evaluation of blood group antigens in ethanol soluble substances (ESS) obtained from human, baboon and vervet monkey red blood cells. Soluble blood group substances, isolated from the red blood cells of humans, baboons, and vervet monkeys by ethanol extraction, possessed serologically active specificities for the following antigens: A, B, H, Lea, LebL, P, P19 Pk and I. Human red blood cells lacking any of these specificities by the direct hemagglutination test also lacked the related antigens in their soluble extract. The only exception was in "Bombay" Oh cells, from which soluble H substance could be readily isolated. Soluble substances obtained from baboon and vervet monkey red blood cells, which lack the human variety of A, B, and H antigens on their red blood cells, inhibited both human and lectin anti-H reagents. The detection of "hidden" H activity in Oh cells will pose some important questions regarding membrane characteristics and the role of immune surveilance.


Antileukocyte antibody in postpartum and renal transplant subjects. A comparison of capillary agglutination and lymphocytotoxicity reactions. Sera were obtained from 48 gravida II prenatal and 211 multiparous nonpregnant females and examined for leukocyte antibodies comparing a standard lymphocytotoxicity (CY) test with capillary agglutination (CA). Antibody was detected in 41% of the samples in both groups but only CA tests were positive with approximately one-half of the prenatal and three-fourths of the multiparous specimens. Although, CA reactions, when accompanied by positive CY responses, usually correlated with HLA, no correlation with HLA, 5b, or the neutrophil antigens was determined for 35 of the 48 sera reacting only by CA. As a model to test the specificity of CA positive-CY negative antisera, four extensively studied sera were further analyzed in 16 families. Independent segregation from the HLA complex and ABO and Rh antigens was confirmed and two of the sera appeared to detect separate clusters of reactions in conjunction with some of the other reagents. Pre- and postgraft samples obtained from 23 living related and 75 cadaveric renal transplanted patients were investigated and compared for graft function and prospective tissue typing. Although direct crossmatches were negative prior to surgery, 17.9% of the pretransplant samples from living related and 28.0% from cadaveric recipients contained detectable antibody when tested against a cell panel. Similar to the prenatal and multiparous groups, the majority of these responses were detected by CA. Following engraftment, antibody first became evident in 11 of 19 (58%) living related and in 23 of 53 (48.2%) cadaveric hosts. There was a striking association between the development of CA and CY antibody and failure, as contrasted to 100% 9-month or greater survival in 10 of 10 living related and 15 of 15 cadaveric transplants in whom only CA antibodies arose postoperatively. In total, these studies indicate that CA reacts with HLA antigens in common with CY tests. In addition, CA may detect HLA when CY is negative but many other reactions appear to be directed at non-HLA specificities. The relevance of CA-only responses to clinical transplantation remains uncertain, but we may speculate that they have an enhancing effect on the course of renal transplantation and are associated with important histocompatibility determinants.


Indian childhood cirrhosis of the liver: a morphologic evaluation. Indian childhood cirrhosis of the liver continues to be a major cause of morbidity and mortality of children living in the Indian sub-continent. The present paper deals with the morphological changes in various stages of the disease process. Ballooning and feathery degeneration of hepatocytes, complete absence of fatty change, frequent occurrence of intracytoplasmic hyaline bodies, dissection of pseudolobules by fibrous septa and fibrosis around single cell were notable features. A few cases also showed giant-cell transformation and pseudo-adenomatous arrangement of hepatocytes. The interpretation of morphologic changes has been discussed and the probability of a toxic aetiology has been suggested.


[Proteinuria in quartan malaria-infected Aotus monkeys]. This paper describes the characterization of proteinuria in Aotus monkeys infected with quartan malaria (Plasmodium brasilianum), using a micro-disc-electrophoresis system. In the post infection urine samples, increases in total proteinuria, albuminuria and gamma-globulinuria were noted a few weeks after peak parasitaemia. Two new proteins also appeared in the urine of malaria infected animals. These findings are discussed with reference to the belief that the Aotus-P. brasilianum system can be a model for human malaria in renal immunopathology.


[Preparation of porcine IgA and its detection using the fluorescent antibody technic]. The preparation of immunoglobulin A (IgA) from porcine colostrum, intestinal content and serum is described. The best results were achieved with colostrum, from which an antigen of satisfactory purity was prepared by purification on Sephadex G-200, on DEAE cellulose and subsequent filtration on Sephadex G-200. The serum to this antigen raised in rabbits was adsorbed to an immunoadsorbent from porcine serum (PS) or porcine IgG. The adsorbtion of the serum against secretory IgA (SIgA) to PS removed its undesirable heterologous and nonspecific reactivity. The anti-SIgA serum adsorbed in this way still reacted with IgA from porcine serum. In the direct and indirect immunofluorescent staining we detected the main antigenic determinants of the SIgA molecule, i. e. the heavy chains and the secretory component.


[Radial scars (scarring) and obliterating mastopathy (author's transl)]. The lesion, known under several names, consists of an hyalinized sclerotic center containing abundant elastic and elastoid masses. These radiate into the periphery and enclose lobuli which reveal epithelial proliferation varying from simple hyperplasia with epithelial villi to the rather rare true papillomas. The hyaline and elastic masses in the central parts surround tubuli and correspond to a lesion known as obliterating mastitis (mastopathy) that may lead to total occlusion of the ducts. The entire process ends with a contracting scar that alters the lobuli stemming from the obliterated duct. These may atrophy, dilate, or respond with different forms of hyperplasia. Therefore the designation "radial scars (scarring) possibly with epithelial proliferation" is proposed. As is emphasized the condition may be associated with tubular carcinoma and may resemble pseudoinfiltration in sclerosing adenosis.


Basement membrane-changes in membranoproliferative glomerulonephritis: a light and electron microscopic study. This study is based on 31 renal biopsies from 28 patients with idiopathic membranoproliferative glomerulonephritis (MPGN). 18 cases were classified as "pure" MPGN, 10 as lobular GN. For light microscopy two staining procedures were found to be of particular value: Pearse trichrome (PAS+Orange G) and Jones-Chromotrope (methenamine silver + Chromotrope 2R). These techniques reveal conspicuous basement membrane (b.m.) lesions which are not observed in other types of GN and are characterized by thickening of the b.m. and a bright orange or red coloration, respectively, which can well be separated from normal (PAS-positive or argyrophilic) basement membranes. Electron microscopy was performed in 13 cases in order 1. to analyze the fine structure of this lesion, 2. to match the results with the current subclassification concept which is essentially based on the discrimination of intramembranous dense (IMDD) and subendothelial deposits. The typical finding of a continuous intramembranous electron-dense material which proves to be argyrophilic in ultrathin sections was present in 3 (23%) of the cases.


[Chemoradiation therapy of inoperable lung cancer]. The results of utilization of different variants of the complex therapy in 416 patients with inoperable lung cancer are reported. 5-fluoruracil, cyclophosphan, methotrexate, chrysomallin were employed as antitumor drugs. Radiotherapy was carried out on a distance gamma machine. The best immediate and early results were gained in patients treated by distance gammatherapy against the background of cyclophosphan injections. The results proved to be most favourable within the terms of 3 years and longer in case of gammatherapy associated with 5-fluoruracil. Application of a fractionated radiation course rendered no positive effect on the results of treatment.


Quantitative Immunological study of pregnancy-associated alpha2-globulin antigen. Pregnancy-associated alpha2-globulin antigen, a protein whigh is practically absent from the serum of nonpregnant individuals or could be found in the quantity of only a few milligram percent, gradually accumulates during pregnancy. A peak value of 96 +/- 44 mg% is reached by the 31st week of gestation. In the last 2 months of pregnancy a decrease was observed to 65 +/- 30 mg% before delivery. Significantly lower levels of this protein could be found in prolonged pregnancy and in case of abortions associated with placenta destruction. Elevated amounts of this characteristical pregnancy protein could be found in 63% of 100 tumor patients.


[Use of various immunological methods for the detection of specific antigen in leukocytes of leukemic patients]. A specific antigen was found in white blood cells of patients with leukemia by means of the indirect immunofluorescence and ring precipitation tests. Using a mixture of hyperimmune rabbit sera with leukocytes from patients with various forms of leukemia (lympholeukemia, myeloleukemia, hemocytoblastosis and reticulosis), it can be found in cells of patients with some forms of this disease. An increased amount of the antigen in the cells was found in cases of myeloleukemia or hemocytoblastosis. The immunofluorescence test was found to be more clear-cut when the time between bleeding and microscopy of preparations was short.


Effect of bleomycin on the fine structure of mouse fibroblasts. Asynchronously dividing mouse fibroblasts (L-cells) treated with the antitumour antibiotic Bleomycin show various rather specific morphological alterations. Many of the cells exposed to bleomycin assume a more or less epitheloid cell shape and are larger than untreated cells; in addition to an increase in nuclear size these cells often contain multiple nuclei. In most of the cells nuclei show an almost complete loss of peripheral condensed chromatin. The nucleolar hypertrophy initially observed is followed by a shrinkage and segregation of the nucleolar components. The cytoplasmic alterations include dilatation of the cisternae of the rough endoplasmic reticulum as well as an increase of free, non membrane attached ribosomes, often arranged in spiral- and rosette-shaped polysomes; they are not specific for bleomycin.


[The effect of drugs on "sino-atrial conduction time" and on sinus-node automaticity in man]. The effect of atropine, propafenone, and disopyramide on sinus node automaticity and "sino-atrial conduction" was tested in normal patients and patients with the sick sinus-syndrome. "Sino-atrial conduction time" was estimated indirectly by the extrastimulus technique. Atropine (n = 11) caused a significant increase in heart rate in all patients. The sinus node recovery time was shortened in 10 patients. "Sino-atrial conduction time" decreased on an average 35% (P less than 0.01). Three patients with a sick sinus-syndrome demonstrated a change of the pattern of the postextrasystolic pauses indicating great improvement in sino-atrial conduction. Propafenone (n = 10) led to a significant prolongation of the sinus node recovery time by 17% and of the "sino-atrial conduction time" by 27%. Disopyramide (n = 8) had no significant influence on heart rate and "sino-atrial conduction time". Sinus node recovery time was not changed in 6 patients. However, in two patients  with a sick sinus-syndrome a dangerous prolongation of the sinus node recovery time after application of disopyramide occurred. The results indicate that atropine enhances sinus node function and sino-atrial conduction. On the other hand, propafenone and disopyramide exert either a depressant influence on sinus node automaticity or on sino-atrial conduction.


[Experimental findings on tyramine-dependent release of noradrenaline in various parts of the heart under the influence of practolol]. Determinations of Noradrenaline-concentrations (NA) in the myocardium of the atria and ventricles of rats were performed and the influences of Practolol and Tyramine were controlled. Following Tyramine, NA was decreased significantly in all specimens. After preinjection of Practolol, NA depletion was considerably less in the right and left atria compared to control values. By these results, the clinical observation of a preferential antiarrhythmic action of Practolol on atrial extrasystoles and tachycardias may be supported.


The biphasic Ca2+-uptake by the fragmented sarcoplasmic reticulum. The non-equilibrium dialysis has been used for kinetic studies of ATP dependent calcium uptake by the sarcoplasmic reticulum. The uptake displays two phases, which are defined as fast and slow uptake. The former is an exponential function of time, with a half-life time of approximately 15--20 sec, the latter presents the characteristics of an autocatalytic reaction. The fast uptake is characterized by its amplitude, the slow uptake by its rate. Compared with the fast uptake, the slow uptake requires higher amounts of Mg2+ or ATP, and is more sensitive to pH variations and aging. The reasons which formerly prevented the resolution of the second phase from the first are discussed. It is concluded that the fast uptake is not a simple binding reaction, and that the slow uptake is more sensitive to changes by the vesicular membrane.


Haemostatic plugs as a histological vital reaction in the skin wounds of guinea pigs. Applying a histological staining method (Carstairs' modification of picro-Mallory staining), haemostatic plugs could be demonstrated at points of vascular lesion in one-minute vital, or older, skin wounds of the guinea-pig on the average in 33% if the specimen was taken immediately after the reaction period, and in about 10% of wounds examined if the specimen was taken five days after death. No haemostatic plugs could be observed in postmortally inflicted skin wounds.


[Cytotoxic effects of a leukocidin from Staphylococcus aureus (author's transl)]. In an attempt to improve the assay of leukocidin from Staphylococcus aureus a combined vital fluorochrome staining procedure with acridine organe and ethidium bromide was developed. It proved to be more suitable for the demonstration of leukocidic effects on leukocytes than phasecontrastmicroscopy. Damaged leukocytes fluoresced distinctly red and undamaged leukocytes green. With the fluorochrome-method leukocidic changes became also evident in some lymphocytes. These were not clearly demonstrated by phasecontrastmicroscopy.


[The influence of brain dysfunction on child development]. In childhood and adolescence two forms of so-called ''psychoorganic syndrome" are distinguished. The early form, in the English and American literature called minimal brain dysfunction, impairs the entire development of the personality in its somatic and psychic aspects. It requires extensive therapeutic efforts. Early diagnosis therefore is of great preventive importance. The infantile psychoorganic syndrome which is acquired later in childhood can be distinguished from the early form by the fact that the impairment is usually only temporary, the etiology is understood and therefore understanding for the child is present. Minimal brain dysfunction when it continues into adolescence is called juvenile psychoorganic syndrome. The emotional liability and irritability which characterises the pubescent period are intensified in the early adolescent with juvenile psychoorganic syndrome and therefore stand out.  The brain damage acquired in adolescence resembles the one acquired in adulthood. It is characterised however by the problems of this age group (identity, vocational decisions, social sexual maturations etc).


[Protein sorption with mineral colloids]. A study was made of the effect on sorption of the molecular weight of model proteins (ribonuclease with a molecular weight of 12 10(3), trypsin with a molecular weight of 24-10(3), bovine albumin with a molecular weight of 64-10(3) and gamma-globulin with a molecular weight of 160-10(3)) and dispersity of suspensions of aluminium hydroxide, aluminum phosphate and calcium phosphate used as biopreparation sorbents. The expediency of using phosphate and calcium phosphate used as biopreparation sorbents. The expediency of using for effective sorption of a definite area of sorption surface necessary and adequate for the distribution of protein macromolecules with the best degree of conformational liberty was revealed.


[Hemagglutinin specificity of Cl. botulinum types A, B, and F in reaction with erythrocytes of various animals]. The following differences were revealed in the haemagglutination reaction with the erythrocytes of man, sheep, rabbit, chicks and mice between the haemagglutinins of Cl. botulinum, types A, B and F, having a close affinity with one another: haemagglutinin of type A actively reacted with the erythrocytes of man, sheep, rabbit, rats and chicks; haemagglutinin of type B reacted only with the erythrocytes of man and rabbits; haemagglutinin of type F failed to react with any of the types of the erythrocytes tested; only with the use of erythrocytes treated with neuraminidase was it possible to establish the presence of haemagglutinin fraction in Cl. botulinum, type F. Treatment of human erythrocytes with neuraminidase and proteolytic enzymes caused a marked increase in the sensitivity of the haemagglutination reaction in haemagglutinins of Cl. botulinum, types A, B and F.


[Dry erythrocytic diagnostic agent for the determination of antiglobulins]. Dry erythrocytic diagnostic agents were obtained under experimental conditions for determination of antiglobulins forming in the organism of man and animals under the effect of serum preparations from the blood of horses and homologoum immunoglobulins. A study was made of the sera of 100 patients with tick-borne encephalitis treated with heterologous and homologous immunoglobulins of directed action; in response to the administration of horse gamma-globulin antiglobulins (in titres below 1 : 10000) appeared in the serum; they circulated in the blood for long periods and inhibited the accumulation of hormonal antibodies to the causative agent; in the majority of cases a high level of antiglobulins to the foreign protein correlated with the presence of remote side-reactions of the serum sickness type. In patients treated with immunoglobulin of human origin antiglobulins were determined in low titres, disappeared from the blood in 15--20 days and did not hinder the accumulation of antihemmagglutinins to the tick-borne encephalitis virus.


[Dynamic aspects of RNA metabolism in the brain of mollusca during electrical effects]. The dynamics of RNA and protein metabolism (as indicated by the incorporation of labeled precursors) during electrophysiological 'learning" and electrical stimulation of isolated ganglia of the snails Tritonia diomedia and Helix pomatia has been investigated. 'Learning" phenomenon manifested itself in the maintenance of synchronous discharges in the electrical activity of the neurons after cessation of the stimulus. At all the electrical stimulations, fast periodic processes of accumulation-degradation of RNA and protein were observed. Radioautographic studies indicate that these processes begin at first in the glia and then in the neurons. The pattern of incorporation of the precursor into the protein is similar to that into RNA, indicating that short-living RNA is involved into the synthesis of short-living proteins. Aurantin (a mixture of actinomycines) inhibits glial synthesis of RNA to a greater extent than neuronal one,-both at rest and electrical stimulation. On the other side, aurantin hinders or even completely blocks "learning" phenomenon. It is suggested that normal RNA metabolism is necessary for the formation of trace phenomena. No essential differences in the metabolism of RNA, protein and nucleotides were found during "learning" and stimulation.


The influence of gramicidin A and valinomycin of the permeability of mammalian erythrocytes. 1. The K+ transport in erythrocytes induced by gramicidin A or valinomycin is a first-order reaction. The activation energy of the induced transport is low and amounts to 6 and 10 kcal/mole, respectively. The indirect method for calculation of the driving force of diffusion, c, is given; in pig erythrocytes the c value for gramicidin A is 21.7, and for valinomycin 35.4 mM-KCl. 2. The valinomycin-induced transport was found to be the same in erythrocytes and biomolecular lipid membranes. The gramicidin A-induced transport corresponds to that of a single gramicidin channel, and not to the multichannel transport observed in the model systems. 3. Erythrocytes of various mammals show large differences in sensitivity to the ionophores. No correlation could be found between membrane lipids and the induced permeability. The role of membrane proteins in ionophore-induced permeability is discussed.


[Retardation of growth and development of rats induced by inhibition of protein synthesis at early postnatal ontogenesis]. Treatment of rats with aurantine for 7-30 days reduces the growth and development of animals, and especially of skeletal muscles. Low relative weight of muscles in aurantine-treated animals was accompanied by low resting and action membrane potentials. Incorporation of labelled uridine and lysine into muscles, heart, brain and liver was decreased. Retardation in the growth and development of skeletal muscles resulted into unfavourable shift of the ratio body weight/surface and led to prevalence of catabolic processes over anabolic ones (increased oxygen consumption, heart and respiration rate in experimental animals). These changes are probably related not only to the inhibition of protein synthesis, but to disturbance of regulatory mechanisms, which reveals itself in an increased norepinephrine content of the brain stem and in the increased cholinesterase activity in cardiac pacemaker.


[Resection of pulmonary metastases (author's transl)]. The authors report on a personal series of 27 patients with pulmonary metastases; seventeen were operated, 3 for cure. Nine hundred and eighty six operated cases from the litterature are then analysed yielding the following notions. Where the primary tumor is controlled at the time of metastatic resection, when there is no systemic or mediastinal dissemination, and when the growth of metastases was slow, these secondary resection of lung metastases were followed by a 16 to 63% 5- year survival rate. The authors conclude it is necessary to control the hypothesis that 5 to 10% of operated cancers present a second operative indication for cure: slow growing metastases limited to the lungs.


A transient rise of hormone secretion: a response of the stimulated rat thyroid gland to small increments of iodide supply. Small doses of iodide (2 times 3.2 mug at 12 h interval), below those capable of inducing Wolff-Chaikoff effect, were injected into rats kept on a moderately low iodine diet. By means of a 125I equilibration technique as well as by direct measurement of cold T4, it was demonstrated that the level of circulating PB125I (representing iodothyronines as confirmed by column chromatography) increased by a mean of 40% within 24 h following the first iodide injection. The serum TSH concentration (measured by radioimmunoassay) was simultaneously depressed. Thus, in stimulated thyroid glands, a biologically significant fraction of an iodide load escapes autoregulatory control of iodothyronine synthesis. A small, transient increase of hormone release is likely to represent the physiological response of a normal gland to a sudden supplement of iodide supply. The ensuing depression of TSH secretion may be necessary for final adjustment of thyroid function. It is considered to be the last step in a cascade of mechanisms whose interaction keeps the thyroidal hormone output within narrow limits in the face of a fluctuating iodide supply. Failure of one or several of these mechanisms in the goitrous human gland could conceivable explain the phenomenon of "Jod Basedow".


Cellular localization of kallikreins in rat submandibular and sublingual salivary glands: immunofluorescence tracing related to histological characteristics. Immunohistochemical localization of rat salivary gland kallikrein was related to glandular structures in tissue processed and stained by various methods. In the submandibular gland, most of the kallikrein was located to cytoplasmic granules of the granular tubules. Cells of the striated ducts showed a faint cytoplasmic staining with a bright luminal rim that occasionally was seen also in the excretory ducts. Minor amounts of kallikrein was found in the interstitial tissue. In the sublingual gland, kallikrein was found in the cytoplasm of the striated duct cells and as a luminal rim in the main ducts. Acini were negative in both glands. Fixation in Helly's fluid preserved cytoplasmic granules and was thus superior for intracellular localization of kallikrein, whereas ethanol fixation, due to absence of non-specific background staining, afforded the most sensitive method for detection of small amounts of antigen. In the submandibular gland, best identification of granular tubules and striated ducts was achieved with DMAB-nitrite staining for tryptophan and counterstaining with Mayer's haemalum on sections of tissue fixed in Helly's fluid. In the sublingual gland, the duct system was best demonstrated by haematoxylin-eosin staining.


Ultrastructure of symmetric interneuronal junctions after staining with ethanolic phosphotungstic acid. 1. The symmetric interneuronal junctions (attachment plaques) of large axon terminals in the dorsal lateral geniculate body of the rat were studied, applying besides the conventional staining the block staining method with ethanolic phosphotungstic acid (E-PTA). 2. After staining these contact zones with E-PTA 3 main components become visible: the interior presynaptic layer, the intermembranous material and the postsynaptic layer. 3. As opposed to the asymmetric synaptic junctions the interior presynaptic layer is not divided into dense projections. 4. The intermembranous material consists of 2 median situated intracleft lines, which are more or less fused with each other. 5. The staining intensity of the paramembranous material of the symmetric junctions stained with E-PTA corresponds to the intensity of the material of the asymmetric synaptic junctions.


Histochemical factors influencing the sulfation of the mucosubstances of the Brunner's glands of the human duodenum. The mucopolysaccharides of the human Brunner's glands were sulfated by sulfuric acid diluted with acetic anhydride or nitrobenzene (1:50) in 5 min. They were recognized by toluidine blue at a pH of less than 1 by exhibiting a red-violet or violet-red metachromasia. The sulfated radicals were removed by methanol or butanol with HCl, H2SO4, Na or NaOH, aqueous Ba(OH)2 at 60 degrees C, or by acetic anhydride, acetyl chloride, benzoyl chloride, or bromine water at 25 degrees C. The carbohydrates were altered so that sulfation was prevented by prior treatment with aqueous Ba(OH)2, or by acetic anhydride with pyridine at 60 degrees C and by acetyl chloride and by bromine water at 25 degrees C. Periodic acid Schiff staining was prevented by sulfation with a nitrobenzene: H2SO4 mixture but not by an acetic anhydride: H2SO4 combination in 1 h suggesting an additional sulfate radical at hydroxyl sites 1 or 2. Brunner's glands contain a large amount of a neutral mucopolysaccharide and can be used as a model for testing a large number of chemical and blockage reactions. In nearly all instances, Brunner's glands reacted more like pyloric glands than duodenal goblet cells.


A procedure for the simultaneous demonstration of neurosecretory and mucosubstances in tissue sections. Many well known histochemical techniques have been employed to study the neurosecretory products in invertebrates and vertebrates. Among these aldehyde-fuchsin (AF) has been one of the two specific stains to demonstrate the presence of neurosecretory cells. Various counterstains have been employed with AF. However, AF is also useful in demonstrating the presence of mucosubstances or elastin. A method is described here for a simultaneous demonstration of neurosecretory substances (NSS) and mucosubstances (MS) by mercurochrome counterstaining. For this method sections of material fixed in methanol-formaldehyde-acetic acid (MFA) or alcoholic Bouin's or Susa were oxidized in 0.3% permanganate with 0.3 ml concentrated H2SO4 for 1 to 2 minutes, decolorized in 1% oxalic acid, stained in 0.5% aqueous basic fuchsin containing 1 ml concentrated HCl and 1 ml paraldehyde for 10 min and counterstained with 0.5% aqueous mercurochrome. After mercurochrome counterstaining the characteristic AF staining (purple colour) of NSS is dramatically swamped (becoming brick red) but the purple developed by MS remains unaltered in vertebrate and invertebrate material. Eosin or other counterstains do not behave in this way.


Alpha-fetoprotein concentration in amniotic fluid during the last trimester in normal pregnancies and in pregnancies with severe fetal abnormalities. The concentration of alpha-fetoprotein (AFP) in the amniotic fluid was studied during the last trimester of pregnancy. 129 samples of amniotic fluid were collected by transabdominal amniocentesis in 94 pregnant women. Only women with uncomplicated pregnancies giving birth to normal infants at term were included. The 90% reference interval was calculated and a distinct decrease in the amniotic fluid AFP concentration was found during the last trimester. An AFP concentration above the 90% reference interval was found in 8 out of 10 cases of anencephaly. Normal AFP concentration was found in a case of congenital heart disease with severe oedema, and a low concentration was found in a case of Down's syndrome (Trisomy 21).


The bound carbohydrates of fractionated serum proteins in protein-calorie malnutrition. Alterations in the carbohydrate composition of serum protein fractions have been determined in pooled sera from 35 children with protein-calorie malnutrition (PCM), 30 children treated for PCM, and 24 age- and social status-matched controls. Total protein-bound hexose (PBH), protein-bound sialic acid (PBSA), and protein-bound fucose (PBF) were elevated in PCM, but only PBF returned to control levels with treatment. Ratios of bound carbohydrate to protein in the unfractioned serum were elevated in PCM but PBH and PBF protein or globulin ratios returned to normal with treatment. The most marked change in carbohydrate composition occurred in the alpha1-globulin fraction. Molar ratios in the individual globulin fractions indicated an increase in carbohydrate moieties as follows: PBSA greater than PBH greater than PBF.


Thin-needle aspiration biopsy. A personal experience with 469 cases. The thin-needle aspiration biopsy technic was used in 469 cases to diagnose benign and malignant tumors. The success of the technic depends upon careful examination of the lesion to be biopsied and attention to the details outlined in procuring the aspiration material and preparing the smears. The most important technical consideration is creating a simulated tissue pattern in the preparation of the smears. Routine Papanicolaou, May-Grünwald-Giemsa, metachrome B, and hematoxylin and eosin stains are all useful. Special stains may provide additional evidence for specific diagnoses. Lymph nodes were the most common tissue examined by aspiration biopsy in this series. It was possible to identify the primary tumor site from the aspiration smear in most of the cases. Forty-seven breast cancers were identified from 127 aspiration biopsies of breast lumps. Breast aspirations are the most difficult to interpret, and no deviation from classic malignant smears should be attempted. The false-negative rate for breast aspirations was 7.0%. No delay in treatment occurred because of a false-negative report. Patient acceptance of the procedure was excellent.


Thyrotoxicosis with painless thyroiditis. Four women and one man with painless subacute thyroiditis presented with hypermetabolic signs and symptoms. Thyroxine (T4) and triiodothyronine (T3) resin uptakes (T3R) were increased but the 24 hour radioactive iodine (RAI) uptakes were less than 1 per cent. Surreptitious use of thyroid hormone was excluded. The thyroid was enlarged in one patient and nontender in all. Exophthalmos was absent. The protein-bound iodine level was 1.1 to 9.5 mug/dl greater than the T4 level. The sedimentation rate was normal or minimally increased, and antithyroglobulin and antimicrosomal antibodies were undetectable. In one hospitalized patient 84 per cent of the administered dose of 131I was recovered in the urine within 48 hours (normal 64 per cent) excluding extrathyroidal uptake. In all subjects the T4 and T3R levels fell to normal or slightly below normal within one to four months. An increase in the 2 and 24 hour RAI uptake to minimally increased or high normal values and return of the T4 and T3R levels to normal occurred in four of five patients within six months. In one of these, the administration of thyroid-stimulating hormone (TSH) resulted in an appropriate increase in 24 hour RAI uptake from 14.9 to 37.1 per cent. One woman remained clinically hypothyroid for six months with a low T4 concentration (3.2 mug/dl), an elevated TSH level (48 muU/ml) and evidence of a persistent organification defect -- two hour RAI uptake decreasing from 33 to 23 per cent after the administration of perchlorate and the 24 hour RAI uptake increasing from 32 to 76 per cent following the administration of TSH. At 21 months after the initial onset of her illness, she is euthyroid but increased RAI uptake persists. The clinical course in four of the five patients is similar to that in an additional eight patients treated during the same time period who presented with typical subacute thyroiditis. Thus, these patients have a form of painless subacute thyroiditis which presents as thyrotoxicosis but is differentiated from it by a low RAI uptake and in whom recovery of thyroidal iodine trapping is the first indicator of recovery. The hyperthyroidism is self-limiting and should be treated conservatively.


Cancer diagnosis. The role of tumor-imaging radiopharmaceuticals. Several radiopharmaceuticals have recently been shown to have a considerable affinity for malignant tissue. All the tumor-seeking radiopharmaceuticals in current use are nonspecific and may also be picked up by benign tumors and infectious processes, including abscess and granuloma. The sensitivity of the tumor-imaging procedure depends on the radiopharmaceutical employed, the type of tumor, its size and location, and previous or current treatment. Gallium-67 citrate (67Ga), the most widely used tumor-seeking radiopharmaceutical, seems to have its greatest value in detecting bronchogenic carcinomas irrespective of cell type. The sensitivity for lung cancer in 489 studies was 93 per cent. Gallium-67 is also of great value in the staging of Hodgkin's disease, in which its sensitivity is 87 per cent. Non-Hdgkin's lymphomas are detected with only slightly lower sensitivity. There is, in fact, evidence that 67Ga is at least complemenatry, if not more sensitive than lymphangiography, in the staging of lymphoma. However, adenocarcinomas originating in the gastrointestinal tract are detected by 67Ga with a sensitivity of only about 40 per cent, whereas various chelates of bleomycin (including 111In-Bleo, 99mTc-Bleo and 57Co-Bleo) detect adenocarcinoma of the gastrointestinal tract with considerably higher sensitivity. In the few studies available comparing bleomycin chelates, 57Co-Bleo and 99mTc-Bleo appear to be more sensitive in detecting tumor than 111In-Bleo. Other tumor-seeking radiopharmaceuticasl which have been employed with somewhat less success include selenium compounds, labeled pyrimidines, several inorganic cations, lanthanide chelates and labeled proteins. Yet to be evaulated clinically is the efficacy of radiolabeled antibodies which are specific for tumor antigens, such as 131I-anti-CEA (carcinoembryonic antigen).


Methods for isolation of anaerobes in the clinical laboratory. Major points to be emphasized include: 1) Anaerobes are very numerous in nature, but only a relatively small number of species are commonly associated with disease in man; 2) The majority of anaerobic bacterial infections involve endogenous rather than exogenous microorganisms, and the patients are usually compromised in some manner prior to infection; and 3) Success in isolating anaerobic bacteria requires careful performance and control of each step in the isolation procedure. These steps include selection and collection of clinical specimens, microscopic examinations, selection and use of primary isolation media, use of anaerobic systems, incubation of cultures and subculture of colonies to obtain pure culture isolates.


Immunoglobulins and beta-1-C globulin in hydatidiform mole vesicles. Qualitative and quantitative determinations of immunoglobulins (Ig's) and beta-1-C globulin were carried out in fluid vesicles of hydatidiform moles and carrier mothers. The mean concentration of IgG was 9.9 per cent, IgA had 14.3 per cent, in relation to maternal values. No IgM concentrations were detected in molar fluid. Beta-1-C globulin showed 60.5 per cent concentration in relation to mother's serum. It is suggested that in trophoblast of molar vesicles a selective transudate migration of Ig's by a mechanism of pinocytosis takes place to the vesicular fluid.


Fine structural observations of the liver in alpha-1-antitrypsin deficiency. Morphologic studies of liver tissue in individuals deficient in alpha-1-antitrypsin (alpha-1-AT) have established the presence of membrane-delimited deposits which are diastase resistant, perodic acid-Schiff positive, sialic acid deficient, and immunologically related to serum alpha-1-AT. The molecular basis for the accumulation alpha-1-AT-like substance in hepatocytes and the serum deficiency in alpha-1-AT patients is unknown. In an effort to gain insight into the membrane sites involved in the storage of the alpha-1-AT-like material, we examined liver biopsies by light and electron microscopy from 3 children with homozygous PiZZ deficiency and varying degrees of liver pathology. Contrary to the more widely held belief that accumulation occurs primarily in the rough endoplasmic reticulum, we find the earliest and greatest accumulation of alpha-1-AT-like material in smooth endoplasmic reticulum of hepatocytes. We have combined our ultrastructural observations with the current knowledge which is available concerning the structural properties of M-type and Z-type alpha-1-AT and have proposed a model which may explain the basis for the hepatic accumulation of alpha-1-AT-like material and the serum deficiency state in the PiZZ genotypes.


Effect of antilymphocyte serum (ALS) on shock in rats. Effects of treatment with rabbit antirat anti-lymphocyte serum and globulin (ALS and ALG) on shock survival were studied in Sprague-Dawley derived male rats. Because of their known cytotoxic capability, it was postulated that lymphocytes might play a role in the pathogenesis of shock and that suppression of lymphocyte function by ALS/ALG treatment should then protect against shock. Shock models used were tourniquet, endotoxin, and hemorrhagic shock. Protection against tourniquet shock was found for ALS made against thymocytes but not for ALS against spleen cells or lymph node cells. The shock-protective factor was found in the ALG-containing serum fraction but not in the primarily albumin fraction. No significant protection was found for ALS treatment against either endotoxin or hemorrhagic shock. ALS effects on blood cell counts, reticulo endothelial system clearance, and inflammation were studied to help identify effects of ALS on shock survival. It was concluded from these studies that thymic or thymus-processed lymphocytes could play a role in the pathogenesis of shock but that multiple effects of ALS/ALG treatment necessitate further studies to elucidate any role for lymphocytes in shock.


[The protective effect of hydroxyethylstarch (HES) in the cryoconservation of bone marrow of the mouse and of man (author's transl)]. The cryoprotective effect of hydroxyethylstarch has been tested in mice and in man using the spleen colony technique (CFU-S test) or the agar colony technique (CFU-C test) as viability assays for the haemopoietic stem cell. In mice we found a mean CFU-S recovery of 88.7%+/-3.9% when a rapid, and 77.8%+/-3.2% when a slow, stepwise dilution method was applied to remove the protective medium (17.5-20% hydroxyethylstarch+20% calf serum) after thawing. In man the rapid dilution after thawing gave a mean CFU-C recovery of 80% but a wide scatter.


[Randomised study on histaminelike side-effects of 5 common plasmasubstitutes in orthopedic surgery (author's transl)]. Preceding anaesthesia 750 randomised patients, sub-divided into 5 different age groups, were given 500 ml of a standard plasma-substitute at a flow rate of 25-30 ml/min. There was direct correlation (p=0.1%) of the frequency of side-effects to the substance used; none, however, to the age or general surgical risk. Side-effects were observed in 21.3% with derivates of gelatin as compared to 3.7% with Macrodex and Plasmasteril combined. The rate of more serious anaphylactoid reactions was 6% with Haemaccel, 1.3% with Gelifundol-S, 0.67% with Macrodex and less than 0.67% with Neo-Plasmagel. The study reveals that serious side-effects may be expected in orthopaedictic patients after application of plasma substitutes. As a prophylactic procedure for those patients we recommend, therefore, the application of these substances only after careful consideration of the indications for their use.


[The influence of hydroxyethyl starch on postoperative alterations of blood coagulation (author's transl)]. The influence of hydroxyethyl starch (HES) on postoperative hypercoagulaemia was examined in a double blind study. One group of 10 patients received 500 ml HES during surgery and on the following three days. A comparable group received the same amount of isotonic saline solution. In some respects, hypercoagulaemia was significantly diminished in the HES-group. The diminution could, however, be mainly explained by the diluting effect of the plasma expander.


[The course of the gamma globulines in septic patients of an intensive care unit (author's transl)]. In 19 septic patients from an intensive care unit, changes were found in the gamma-globulin-fractions with the use of the quantitative immunelectrophoresis. This technique of study presents a great improvement over the diagnostic methods which were used previously. In addition, the possibility is offered to treat the detected deficiencies in certain Ig-fractions and to provide a more directed treatment with antibiotics.


Reversible inhibition of rapid axonal transport in vivo by lidocaine hydrochloride. Rats were given standardized injections of 3H-L-proline into the trigeminal ganglion and 14C-lidocaine hydrochloride at the infraorbital foramen. The 3H-L-proline was always injected 2.5 h before removal of the nerve. Lidocaine, 1, 2, and 4%, produced a concentration-related inhibition of entry of 3H-labeled rapid axonal transport into the distal portions of the nerve. Addition of epinephrine, 1:200,000, doubled the intensity of the effect. The time delay of recovery was also concentration-related, and with 4% lidocaine recovery still seemed incomplete after 4.5. h. It is concluded that inhibition of rapid axonal transport is probably a usual byproduct of nerve block with local anesthetics such as lidocaine. The inhibition seems attributable in part to a disturbance of the energy metabolism of the nerve.


Gene dosage and antigenic expression on the cell surface of bovine erythrocytes. Electron microscopic and serological techniques have been used to study the relationship between cell surface expression of bovine erythrocyte antigens and the genes coding for these antigens. Using cells which are genetically and serologically defined for their zygosity with respect to the Z allele, it was found that homozygous (Z/Z) cells have approximately twice as much surface Z antigen as heterozygous (Z/-) cells. Cells labeled for the J antigen, a soluble serum substance which secondarily adsorbs to the erythrocyte surface, display a quantity of antigen which is directly related to the J titer of the cells. A new antigen is described which is independent of the J antigen, and which is detectable by EM labeling and by indirect agglutination, but not by hemolysis.


[Immunochemical study of glomerular basement membrane in human and experimental pathology]. After a review of the histology and the biochemical structure of the glomerular basement membrane, the authors discuss the role of its diverse antigenic components in the origin and course of several types of glomerulonephritis. In addition, they deal with the problem of biochemical modifications of glomerular basement membrane during some experimentally induced or naturally occuring pathological states.


Orcein positive hepatocellular material in histological diagnosis of primary biliary cirrhosis. Liver biopsies from 18 patients with primary biliary cirrhosis (PBC) and from 25 patients with chronic active hepatitis (CAH) were stained by orcein after oxidation of the tissue sections with potassium permanganate. In 15 out of the 18 cases which could be classified on clinical, biochemical and immunological basis as PBC, the hepatocytes, usually periportally, contained cytoplasmic stainable material. 5 out of the 25 CAH patients contained the same material, but four of these patients were clinically atypical and showed features of cholestatic form of CAH and features crosslinking with PBC. All patients in both groups were serologically hepatitis-B antigen negative. The orcein staining method seems to be a reliable addition to differentiating histologically between PBC and CAH.


Immunochemical localization of in vivo bound immunoglobulins in pemphigus vulgaris epidermis. Employment of a peroxidase-antiperoxidase multistep technique for light and electron microscopy. A multi-step immunocytochemical method utilizing horseradish peroxidase (HRP) as an immunological marker was employed to demonstrate, at the ultrastructural level, IgG antiepithelial autoantibodies bound in vivo to pemphigus epidermis, and circulating IgG antibodies, using monkey oesophagus as substrate. To demonstrate IgG the following antisera were employed in sequential steps: Goat antihuman IgG; rabbit antigoat IgG; goat-anti-HRP-serum. After incubating with the final antigen, HRP, the latter was visualized with a cytochemical method. Results paralleled those obtained by immunofluorescence, the antiepithelial antibody being demonstrated at the sites of the intercellular spaces of the epidermis. Electron microscopic cytochemistry showed IgG in the surface coat of epidermal cells and in the intercellular space, both in the desmosomal and the interdesmosomal areas. These results confirm the results of a previous study but the localization of the antibody is more exact than that obtained with HRP-conjugates. The present method is more versatile and specific than HRP methods that utilize HRP-conjugated antibodies.


[The mechanism of antigen-antibody reaction at the basement membrane zone in bullous pemphigoid. The question of interaction of complement factors and bivalent cations (author's transl)]. 1. In a medium poorly cationic indirect immunfluorescence of the basement membrane zone normally observed with preparations of sera from patients with pemphigoid has not be seen. 2. The fluorescence of basement membrane zone takes place if Fe2+ ions are added together with the antibasement zone antibodies. Evidently Fe2+ ions are essential for fixing antibodies. 3. With the method of indirect fluorescence the C3 component of complement could be observed after fixation of pemphigoid antibodies to the basement membrane zone. 4. EDTA prevents binding C3 too. Only after adding Fe2+ (not with Ca2+ or Mg2+) complement IF staining is demonstrable. Apparently EDTA causes the loss of fixing antibasement zone antibodies at first. Thereafter complement staining reaction is negative. 5. Participation of C1 could not be demonstrated using indirect IF staining. The alternate pathway to activation of C3 is discussed.


Complement in pemphigus vulgaris and other bullous dermatoses. The behavior of the complement  and of its components C4 and C3 has been studied in serum and in blister fluid both in patients with pemphigus vulgaris and other bullous dermatoses and in healthy subjects with experimentally induced blisters. The results are suggestive of local activation of the complement, in the blister fluid of patients with pemphigus vulgaris, pemphigus erythematosus and bullous pemphigoid according to the classical enzymatic sequence.


Studies on stimulation and properties of antiglobulin antibodies. III. Antiglobulin antibodies with different specificity in patients with bronchial asthma complicated by suppurative catarrh of the respiratory tract. Sera from 14 of 50 patients suffering from bronchial asthma complicated by suppurative catarrh of the respiratory tract contained antibodies to homologous and heterologous gamma-globulins. The findings were attributed to hyperimmunization causing changed in antigenic properties of antibodies. Antigenically altered antibodies can act as autoimmunogenic stimuli inducing an immune response manifested by the appearance of antibodies to autologous gamma-globulins and cross reacting with homologous and heterologous gamma-globulins.


A trial of utilizing antibodies as carriers of alkylating agents. I. Influence of cyclophosphamide and chlormethine on antigenic properties of serum proteins. Electrophorograms of human and rabbit sera before and after incubation with various concentrations of cyclophosphamide and chlormethine (Nitrogranulogen) were compared. Sera of rabbits injected with chlormethine and cyclophosphamide in therapeutic doses for humans were examined for antibodies reacting with homologous and antologous gamma-globulins. Immunoelectrophorograms of sera incubated with both alkylating agents differed from those of preincubation sera with regard to number and intensity of precipitin lines. The changes in electrophorograms were related to the type and concentration of alkylating agent. No antibodies to homologous or autologous serum antigens of rabbits treated with chlormethine or cyclophosphamide were found.


Marrow adipose cells. Histochemical identification of labile and stable components. Two histochemically distinct populations of fat cells are present in the bone marrow with a topographic distribution paralleling that of hemopoiesis. Fat cells of red marrow stain positively with performic acid-Schiff (PFAS) whereas those of yellow marrow do not stain with this technique. When hemopoietic tissue expands in response to experimentally induced hemolysis, PFAS-positive cells of red marrow disappear whereas the PFAS-negative cells of yellow marrow remain stable. This differential staining indicates a difference in lipid composition of fat cells in red and yellow marrow and may relate to their relative stability during the expansion of hemopoiesis.


Recurrent immunogenic interstitial keratitis. Production in the guinea pig by extraocular means. Combinations of intracutaneous (IC) and intravenous (IV) injections of bovine gamma-globulin (BGG) or bovine serum albumin (BSA) induced interstitial keratitis in 24 of 29 guinea pigs. For IC injection, antigen was incorporated in complete Freund adjuvant. When IC and IV BGG reactants were administered simultaneously, corneal opacities appeared after 11 to 19 days and persisted for periods of 2 to 11 days. Following a free interval, a secondary reaction of somewhat shorter duration occurred. A BGG-adjuvant emulsion administered three or five weeks after IV injection of BGG induced an accelerated and intensified response followed by a protracted course of recurring corneal opacities. Irritative brain lesions produced by implanted alumina cream or cobalt powder prior to simultaneous IC and IV antigenic treatment were associated with diminished conreal reactions (alumina) and a number of tertiary eye responses.


[Estrus synchronization in cattle using chlormadinone acetate ("Bovisynchron" Jenapharm) in the tropics. 1. On the effect of blocking the cycle using chlormadinone acetate and on the introduction of this biotechnical method in the Republic of Mali]. The oestrus synchronization in cattle is being tested as a biotechnical method supporting the implementation of the crossing programme in the Republic of Mali Using the latest results of sexual physiology, the effect is described of Bovisynchron Jenapharm (chlormadinonacetate). Bovisynchron is characterised by a high certainty of synchronization and a good toleration. It leads to a highly fertile oestrus of all animals and to a high rate of conception. An analysis of the situation in the Republic of Mali is used to discuss the advantages of this method.


Antigenic determinants of hen egg-white lysozyme in delayed hypersensitivity. I. Macrophage migration inhibition activities of lysozyme fragments. Five kinds of fragments of hen egg-white lysozyme (HL) were tested by macrophage migration inhibition (MMI) assay using peritoneal exudate cells (PEC) of guinea pigs immunized with HL in complete Freund's adjuvant. (See article). These four kinds of HL fragments were also shown to be composed of the immunodominant groups of the HL molecule for circulating antibody against HL in guinea pigs. The relationships between the antigenic sites related to circulating antibody and the cellular recognition sites are discussed.


Antigenic determinants of hen egg-white lysozyme in delayed hypersensitivity. II. Antigenicity and immunogenicity of the N- and C-terminal peptide. Various small fragments of (see article) which is one of the immunodominant groups of hen egg-white lysozyme (HL), were tested for macrophage migration inhibition (MMI) of peritoneal exudate cells (PEC) from guinea pigs immunized with HL. P17 was split in the middle with cyanogen bromide. The terminal portion of (see article) showed positive MMI, whereas the non-terminal half of P17, P17i (sequence 13-27) only showed very weak MMI activity. A fragment derived from the middle portion of P17, P17m (sequence 11-22), was inactive. When P17 were reduced and alkylated, one of the resultant peptides, P17N (sequence 1-[CM-Cys-6]-27) still has MMI activity with PEC taken from guinea pigs immunized with HL, although no antibody reacting with it was detected, but P17C (sequence 123-[CM-Cys-127]-129) was inactive. The peptides P17 and P17N were both immunogenic in guinea pigs in respect to the delayed hypersensitivity response. Again P17t and P17N were immunodominant groups, but the reactivity of P17i in MMI assay of this group of animals was greater than that in guinea pigs immunized with HL. The reactivities of HL with PEC taken from guinea pigs immunized with P17 or P17N were generally weaker than those of the antigens used for immunization.


The radioimmunoassay of steroid glucuronides. The oestrogen C-3 glucuronides as haptens. Antisera were prepared against three related oestrogen ring-A glucuronides, oestrone 3-glucuronide, oestradiol 3-glucuronide and oestriol 3-glucuronide. The corresponding 6,7-3H-labelled conjugates were synthesized as radioligands and the cross-reactions of the antisera against ring-A oestrogen glucuronides and other steroid conjugates were examined. The specificity of the antiserum against oestriol 3-glucuronide was compared with that raised against oestriol 16alpha-glucuronide, and the measurement of the former conjugate in late-pregnancy urine is discussed.


The comparative specificity of 3 oestradiol-binding proteins. Rat alpha-foetoprotein, rat liver 17beta-hydroxy steroid dehydrogenase and anti-(oestradiol-6-carboxymethyloxime-bovine serum albumin) antiserum. 1. The specificity of 3 oestradiol-binding proteins was studied. Two of these proteins are naturally occurring (rat alpha-foetoprotein and rat liver microsomal 17beta-hydroxy steroid dehydrogenase) and the third is an artificially induced model, anti-(oestradiol-6-carboxymethyloxime-bovine serum albumin) gamma-globulins. 2. A specific binding procedure for each protein model permitted a determination of its affinity for oestradiol and for 30 other steroids. 3. The results obtained have brought to light the different areas of the steroid molecule that are important for its recognition by each of the three proteins. The two naturally occurring proteins (alpha-foetoprotein and 17beta-hydroxy steroid dehydrogenase) recognize the edge of the steroid defined by C-4, C-6, C-8 and C-15. On the other hand, the gamma-globulins recognize the opposite edge, i.e. that defined by C-2, C-10, C-11 and C-17. 4. Diethylstilboestrol, whose structure is analogous to that of a steroid, is only recognized by the two naturally occurring proteins.


The polypeptide composition of bovine epidermal alpha-keratin. 1.  The polypedtide chains that comprise the subunits of the tonofilaments, or th alpha-keratin component, of bovine epidermis were fractionated by combination of chromatography on DEAE-cellulose and preparative polyacrylamide-gel electrophoresis.  2.  The seve polypeptide chains investigated had generalyy similar properties; all contained two residues per molecule of tryptophan and N-acetylserine was the common N-terminal amino acid residue.  3.  On the basis of close similarities in alpha-helix content and amino acid composition, the polypeptide chains were classified into three distinct groups.  Each group contained approximately one-third of the total polypeptides on a molar basis.  The groups and designated polypeptides chain numbers were: group one, polypeptides 1a and 1b, which had moleculae weights of 58,000, contained about 25% alpha-helix, 86 glutamic acid and 8 cysteine residues per molecule, but which differed in net charge, extinction coefficients and tyrosine contents; group two, polypeptides 2, 3, and 4, which hadmolecular weights within thewithin the range of 52,00-56,000, contained about 48% alpha-helix, 54 glutamic acid and 6 cysteine residues per molecule, but which differed in extinction coefficients and tryosine contents; and group, polypeptides 5  and 6, which had molecular weights of 47000-48000, contained about 56% alpha-helix, 64 glutamic acid and 4 cysteine residues per molecule, but which differed in extinction coefficients and tyrosine contents...


Rabbit collagenase. Immunological identity of the enzymes released from cells and tissues in normal and pathological conditions. 1. The immunological cross-reactivity between rabbit collagenases from a variety of normal and pathological sources was examined. The specific antibody raised against collagenase secreted from normal rabbit synovial fibroblasts gave reactions of complete identity with collagenases secreted from fibroblasts derived from rabbit skin, and from synovium from experimentally arthritic rabbits. 2. The rabbit fibroblast collagenase was immunologically identical with collagenases obtained from the organ culture medium of normal rabbit skin, synovium, ear fibrocartilage and subchondral bone. 3. Collagenases from the culture media of normal rabbit synovium and from hyperplastic synovium of rabbits made experimentally arthritic were identical. 4. The collagenase secreted from rabbit fibroblasts gave a reaction completely identical with that of a collagenase extracted directly from a rabbit carcinoma. 5. IgG (immunoglobulin G) from a specific antiserum to rabbit fibroblast collagenase was a potent inhibitor of the collagenases obtained from the culture media of the various rabbit cells and tissues. 6. Collagenases from human synovium and from mouse macrophages and bone were neither precipitated nor inhibited by antibodies to rabbit collagenase. 7. No immunoreactive material was found in lysates of rabbit polymorphonuclear leucocyte granules with the specific antisera to rabbit fibroblast collagenase. No evidence for inactive forms of rabbit collagenase in lysates of the rabbit synovial fibroblasts could be found, either by double immunodiffusion against the specific collagenase, or by displacement of active enzyme from inhibition by the IgG.


Purification of the Thy-1 molecule, a major cell-surface glycoprotein of rat thymocytes. The Thy-1-molecule, which was identified by its antigenic activities, has been purified from rat thymocytes. The purification involved preparation of crude membranes and solubilization in deoxycholate, followed by gel filtration and affinity chromatography on antibody or lectin columns. In all cases the purified molecule was a glycoprotein that did not form higher polymers and was not associated with other polypeptide chains. The Thy-1 glycoprotein could be found in two forms, one binding to lentil lectin, the other not. Both forms had the same detectable antigens and were of a similar but not identical size. After sodium dodecyl sulphate-polyacrylamide-gel electrophoresis the apparent molecular weight of Thy-1 binding to lentil lectin was 25 000, whereas that not binding to the lectin was 27 000, with heterogeneity towards forms of apparently higher molecular weight.


Purification of the Thy-1 molecule from rat brain. The present paper describes the characterization of the Thy-1 molecule from rat brain. The molecule was recognized by its antigens, which could be solubilized from brain membrane with deoxycholate. In the solubilized form the Thy-1 antigens were associated with a homogenous component with the following hydrodynamic properties: s20,w=2.2s,v=0.72ml/g and Stokes radius=3.0nm. The mol.wt. of the deoxycholateantigen complex was estimated to be 27000; these values are not significantly different from those obtained thymocyte Thy-1. Brain Thy-1 was further purified by affinity chromatography with lentil lectin coupled to Sepharose 4B, and more than 80% of the antigen was bound. The material eluted with methyl alpha-D-glucopyranoside was then filtered on a column of Sephadex G-200, and only one glycoprotein was found in the antigenically active fraction. On sodium dodecyl sulphate-polyacrylamide-gel electrophoresis the glycoprotein was very similar to the Thy-1 from thymocytes that binds to lentil lectin. Its apparent mol.wt. on 12.5% acrylamide gels was 24000, and it electrophoresed as a symmetrical band. Brain Thy-1 was antigenically indistinguishable from thymocyte Thy-1 when analysed with rabbit antisera raised against brain or thymocyte Thy-1.


Pharmacological interactions between ditazole and anticoagulant drugs. 4,5-Diphenyl-2-bis(2-hydroxyethyl)aminoxazole (ditazole) 200 mg/kg administered orally for 3 days significantly reduced the one-stage prothrombin time in rats treated concomitantly with bishydroxycoumarin (BHC) or warfarin, orally or i.p., as compared with rats given only BHC or warfarin. No significant change occurred in heparin anticoagulant activity when it was administered to rats pretreated with ditazole for 3 days. Warfarin and BHC, in vitro or in vivo, did not influence platelet aggregation or the antiaggregating activity of ditazole. The co-administration of heparin and ditazole enhanced the antiaggregating activity of the two drugs in both in vivo and in vitro tests.


Histamine release in human subjects by modified gelatin (Haemaccel) and dextran: an explanation for anaphylactoid reactions observed under clinical conditions? Histamine release by modified gelatin (Haemaccel) and dextran (Macrodex) has been demonstrated in volunteers by direct and indirect methods. In a pilot study of Haemaccel, histamine release was observed in six of seven volunteers. The highest plasma histamine concentration was 4.8 ng/ml, the lowest 1.7 ng/ml: two of the subjects showed slight allergic reactions. Using Haemaccel batch 2551, 10 out of 12 subjects reacted to the rapid infusion of Haemaccel with increased plasma histamine concentrations, whereas none reacted to Ringer's solution. None of the 10 subjects had an allergic reaction, but an increase in gastric secretion was observed in eight. Changes in the venous basophil granulocyte count were found in both those who reacted and those who did not react to Haemaccel. After the rapid infusion of dextran the highest plasma histamine concentration was 5.0 ng/ml, the lowest 1.3 ng/ml. The withdrawal of blood had no influence on plasma histamine concentration. The incidences of histamine release produced by Haemaccel varied with different batches. Thus, it seems unlikely that immunological mechanisms are principally responsible. Nine instances of allergic and anaphylactoid reactions to plasma substitutes have been reported, seven after Haemaccel infusion, and two after dextran administration. One of the patients who received dextran died. Histamine release was always associated with Haemaccel infusion and corresponded in extent to the clinical symptoms observed, but there was no significant histamine release associated with the reactions to dextran.


Studies on the structure and immunological activity of carcinoembryonic antigen - the role of disulphide bonds. Carcinoembryonic antigen (CEA) has been shown to contain no free cysteine thiol groups but 6 cystine disulphide bonds. 5'5-Dithiobis-(2-nitrobenzoic acid) (DTNB) will react with CEA only after reduction of the disulphide bonds with dithioerythritol. Reduction-alkylation of CEA using dithioerythritol and bromo-[1-14C] acetic acid confirmed the presence of 6 disulphide bonds, as did oxidation of the glycoprotein with performic acid. The products from the DTNB and reduction-alkylation treatments of CEA had less capacity to inhibit the binding of [125I]-CEA to anti-CEA in a radioimmunoassay than the original CEA but could, in sufficient quantities, totally inhibit the binding. Removal, using mercaptoethanol, of the thiol blocking groups from the DTNB-treated CEA resulted in a 55% recovery of antigenic activity. The product from the performic acid oxidation could only inhibit approximately 50% of the binding. Treatment of CEA with 0.533M sodium periodate (NaIO4) greatly reduced its antigenic activity, presumably a result of the oxidative cleavage of the disulphide bonds. No loss in activity, however, was observed when 5.33mM NaIO4 was used, and one Smith degradation (i.e. treatment in sequence with periodate, borohydride and mild acid) of CEA removed approximately 50% of the carbohydrate, including all of the fucose, sialic acid and 2-acetamido-2-deoxygalactose but did not change the antigenic activity.


Characterization of tumour cell aggregation promoting factor from rat ascites hepatoma cells: Separation of two factors with different antigenic property. The previously described glycoprotein that promotes tumour cell aggregation, derived from rat ascites hepatoma cells and capable of partial purification by chromatography, was found to be a mixture of 2 factors with different antigenic property. One was not absorbed by immunoadsorbent chromatography with anti-rat serum antibody and the other was. The action of the unabsorbed factor was clearly more potent than that of the absorbed factor. Both the factors were found in the serum of tumour bearing rats and the action of the unabsorbed factor was also more potent than that of the absorbed factor; its amount increased with time after i.p. inoculation of the cells. The serum of healthy rats contained the absorbed factor but not the unabsorbed factor. It was thus assumed that the unabsorbed factor was associated with the hepatoma cell surface itself and released into the serum, while the absorbed factor was associated with serum protein coating the cell.


Maternal plasma alpha-fetoprotein in normal and complicated pregnancies. Plasma alpha-fetoprotein (AFP) has been estimated by radioimmunoassay in healthy non-pregnant adults, some of whom were taking oral contraceptives, and in women with normal and complicated pregnancies. A curve with the 90 per cent range in normal pregnancy was established. The prop&rtion of values below the 5th centile in complicated pregnancies was significantly greater than in uncomplicated pregnancies. However, three-quarters of the estimations in the complicated groups lay within the 90 per cent range. Nor correlation was found between maternal plasma AFP values and the incidence of fetal distress, small-for-dates babies or perinatal mortality. It is concluded that the measurement of maternal plasma AFP is not reliable test for predicting fetal well-being.


Elevated levels of maternal plasma alpha-fetoprotein after amniocentesis. An elevation of maternal AFP levels was observed in 11 of 65 cases (17 per cent) after amniocentesis. It is suggested that blood samples in which AFP levels are to be measured should always be collected before and not after amniocentesis.


Maternal serum alpha-fetoprotein and previous neural tube defects. A case-control study involving 63 pregnancies showed that women who had previously had an infant with anencephaly or spina bifida did not have raised serum alpha-fetroprotein (AFP) levels in subsequent unaffected pregnancies. The value of a serum AFP determination in such women is discussed. If the serum AFP level is normal and if ultrasonography excludes anencephaly the risk of spina bifida might be low enough (about 1 per cent) to make diagnostic amniocentesis difficult to justify. Conversely, a high serum AFP value in such women should not be acted on without confirmation of an abnormality by ultrasonography and, if that is negative, by diagnostic amniocentesis.


Serum protein fractions from children of differing nutritional status analysed by polyacrylamide gel electrophoresis and electroimmunoassay. 1. The nutritional status of children showing no clinical signs of malnutrition, from the University School of Khon Kaen, Khon Kaen province, north-east Thailand and from two villages nearby, was tested. The children were grouped according to their body-weight expressed as a percentage of expected weight-for-height (Harvard standards (Stuart & Stevenson, 1959), as given by Jelliffe (1966)). 2. The differing prealbumin concentrations indicated that nutritional status differed between the groups. 3. The urinary urea: creatinine ratio was significantly lower in the village children compared with the children from Khon Kaen, indicative of the higher dietary protein intake of the latter. 4. alpha1-Acid glycoprotein and the first 'post-albumin peak' (obtained by polyacrylamide gel electrophoresis of serum and containing mainly mainly Gc-globulin, alpha1-antichymotrypsin and alpha1-B-glycoprotein) were found to be significantly higher in the village children compared with children from Khon Kaen. 5. The three main proteins of the first 'post-albumin peak' from polyacrylamide gel electrophoresis of serum were tested separately using the electroimmunoassay method. There was no significant difference in Gc-globulin between the children from Khon Kaen and the village children. The concentration of alpha1-B-glycoprotein from those Khon Kaen children whose body-weight was more than 95% expected weight-for-height was significantly lower compared with that of village children, alpha1-Antichymotrypsin concentration was significantly higher in serum from Khon Kaen children than in serum from village children.


Studies on the isolation and partial characterization of apolipoprotein D and lipoprotein D of human plasma. This report describes further studies on the characterization of apolipoprotein D (ApoD), a recently recognized human plasma apolipoprotein, and presents results on the isolation and distribution of its lipoprotein form, lipoprotein D (LP-D). ApoD, isolated by a procedure combining hydroxylapatite and Sephadex G-100 column chromatography, migrated on 7% polyacrylamide gel as a single band with a mobility intermediate between those of A-II and C-II polypeptides. On double diffusion and immunoelectrophoresis, ApoD reacted only with antiserum to ApoD. It was characterized by the presence of all common amino acids including half-cystine. The amino terminal acid was blocked. Carbohydrate analysis demonstrated that ApoD is a glycoprotein with glucose, mannose, galactose, glucosamine, and sialic acid accounting for 18% of the dry weight of ApoD. The estimated molecular weight of ApoD IS 22 100. ApoD occurs in the serum as a lipoprotein which was isolated from high density lipoproteins3 by two different chromatographic procedures. In the first procedure, high density lipoproteins3 were treated with neuraminidase and chromatographed on concanavlin A. The retained fraction containing LP-D was purified by hydroxylapatite column chromatography. Alternatively, LP-D was isolated by a procedure combining chromatography of high density lipoproteins3 or whole serum on an immunosorber containing antibodies to ApoD, and hydroxylapatite column chromatography. LP-D displayed a single, symmetrical boundary in the analytical ultracentrifuge and a single band on 7% polyacrylamide gel electrophoresis. When injected into rabbits it produced antisera that reacted only with ApoD. On immunoelectrophoresis LP-D had a mobility different from that of lipoprotein A (LP-A). A direct immunological comparison of LP-D and LP-A showed a reaction of nonidentity. LP-D consists of 65-75% protein and 25-35% lipid. The lipid moiety contains cholesterol, cholesterol ester, triglyceride, and phospholipid. The phospholipid. composition is characterized by a relative high content of lysolecithin and sphingomyelin and a relatively low content of lecithin. We have concluded from these studies that ApoD is a unique apolipoprotein that exists in the form of a distinct lipoprotein family with a macromolecular distribution extending from very low density lipoproteins into very high density lipoproteins, but with a maximum concentration in high density lipoproteins3 and a minimum concentration in high density lipoproteins.


Studies on the receptors of the MNSs group system. Results with modified human red cell membrane sialoglycoproteins indicate that alkali-labile sialic acid and amino groups are parts of the erythrocyte receptor sites recognized by common rabbit and human anti-M and -N sera. The "N" antigen, demonstrable in MM glycoprotein preparations by rabbit anti-N, has structural properties which are similar to those of the MN receptors. Sialic acid, amino groups and carbohydrate, susceptible to periodate oxidation, are not involved in the Ss antigen sites. The specificity of the Vicia graminea lectin is dependent on free amino and carboxyl groups. Its affinity for the substances is increased by blocking of amino groups.


The homeless person and the psychiatric services: an Edinburgh survey. A representative sample (R.S.) of 79 subjects living in Edinburgh common lodging houses was compared with a clinical series (C.S.) of 44 patients from the same type of resisence. C.S. patients were more likely to be out of work, to be under 55 years of age, and to have been married as some time. They had spent much shorter times in lodging houses, in Edinburgh and at their current address. Alcoholism was rather more often diagnosed in the C.S., and personality disorder much more often. Schizophrenia tended to be found more in the R.S. The C.S. obtained higher Personal Illness and 'Character Disorder' scores. It is concluded that those subjects presenting to the psychiatric services are a highly selected group quite unrepresentative of homeless single persons in general.


Ramification patterns of ganglion cell dendrites in the retina of the albino rat. Ganglion cell morphology has been analyzed in the retina of the adult, albino rat by the Golgi technique and evidence has been obtained that the ganglion cells of this species show a far greater variety and complexity of organization than described previously. Both diffuse ganglion cells, whose dendrites ramify throughout the inner plexiform layer, and stratified ganglion cells, whose dendritic branching is restricted to one or two planes, have been identified. Evidence is also presented for distinct stratification of the inner plexiform layer in this species, based on radioautographic analysis following intravitreal injection of tritiated amino acids.


Projections from the 'cingular' vocalization area in the squirrel monkey. In 5 squirrel monkeys the anatomical projections from the 'cingular' vocalization area were studied by the autoradiographic tracing technique. The 'cingular' vocalization area lies around the sulcus cinguli at the level of the genu of the corpus callosum; its electrical stimulation yields purring and cackling calls. The following efferent connections were found: corticocortical fibers could be traced into the orbital cortex (areas 10 and 11), dorsomedial frontal cortex (areas 9, 8 and 6), limbic cortex (areas 25, 24 and 23), Broca's area (area 44), frontal operculum (area 50), insula (areas 13 and 14), and auditory association cortex (area 22). Subcortical terminal fields within the telencephalon were found in the nucleus caudatus, putamen, claustrum, globus pallidus, olfactory tubercle, preoptic region and nucleus centralis and basolateralis amygdalae. Fibers reached most of these structures along different trajectories. In the diencephalon terminal fields lay in the dorsal hypothalamus, the subthalamus, lateral habenular nucleus, and the following thalamic nuclei: nucleus reticularis, ventralis anterior, centralis medialis, centralis superior lateralis, centralis inferior, submedius, medialis dorsalis and centrum medianum. In the midbrain, the periaqueductal gray was the only projection area, extending into the parabrachial nuclei at the pontomesencephalic transition. The most caudal terminal field was found in the medial pontine gray. No terminals were detected in the nucleus ambiguus, nucleus n. hypoglossi or in any other cranial motor nucleus involved in phonation processes. A comparison of this projection system with the whole of structures producing vocalization when electrically stimulated yielded only partial overlap. Not all vocalization areas lie within the 'cingular' projection system, and inversely, not the whole projection system yielded vocalization. Overlap took place in the anterior limbic cortex, preoptic region, central amygdaloid nucleus, midline thalamus, dorsal hypothalamus, periaqueductal gray and parabrachial nuclei. These structures are considered to compose a functionally coherent vocalization system. The projections into Broca's area, nucleus ventralis anterior thalami, frontoopercular cortex within the lateral fissure, pontine nuclei and superior temporal gyrus are discussed in their possible relationship to vocalization processes.


Antigenic and morphologic comparisons of Ibaraki and bluetongue viruses. Bluetongue disease virus, type 10, and Ibaraki disease virus, which causes a bluetongue-like disease of cattle, were compared to determine whether they are the same or different viruses. They were similar in morphology, but neutralization tests, complement-fixation tests, and ferritin tagging indicated that they have antigenic differences. Therefore, they should be considered as different viruses. Two other viruses of this group, African horsesickness and equine encephalosis, were included to show that Ibaraki and bluetongue had developmental morphological features that could be used to differentiate them from the two equine viruses.


Ultrastructural studies of the nucleoids of the pleomorphic forms of Chlamydia psittaci 6BC: a comparison with bacteria. The nucleoids of the various pleomorphic forms of Chlamydia psittaci have been examined by direct observation of infected cells and by observations on isolated particles. The fixation and staining methods used were the same as those routinely used for the examination of bacteria to facilitate the comparison of chlamydial fine structure with that of bacteria. The nucleoids of reticulate bodies were composed of fine fibrils which extended throughout these particles. The nucleoids of intermediate bodies are characterized by an electron-dense mass with which the fibrous elements are associated in a structurally coherent manner. As condensation of the intermediate bodies proceeds, the electron-dense mass becomes eccentrically located and the fibers form a distinct radiating structure. Large elementary bodies have a few fibers associated with their condensed electron-dense nucleoids but the more condensed mature elementary bodies have a very discrete and homogeneous electron-dense nucleoid which is separated from the cytoplasmic elements of these particles by a very distinct electron-transparent space. These highly condensed elementary body nucleoids are usually ovoid, but may be elongated or irregular, and a small number of these structures react very strongly with ruthenium red. While the nucleoid structure of reticulate bodies resembles that of the bacterial cell, both the condensation process and the nucleoid morphologies which result from it in intermediate and elementary bodies have no parallels among the bacteria. Thus we conclude that major differences in nucleoid organization exist between the chlamydia and the bacteria.


Production of beta-(4-hydroxyphenyl)ethanol and beta-(4-hydroxyphenyl)lactic acid by Candida species. Two crystalline compounds were isolated from the culture filtrates of Candida species grown in synthetic medium supplemented with L-tyrosine as the sole source of nitrogen. These compounds were characterized as beta-(4-hydroxyphenyl)ethanol (HOPEA) and beta-(4-hydroxyphenyl)lactic acid (HOPLA). The production of these compounds in five species (both pathogenic and non-pathogenic) was compared and marked differences were revealed. Experiments using L-[14C]tyrosine indicated that both HOPEA and HOPLA are synthesized from L-tyrosine.


Chemotherapy of germ cell tumors of the testis. I. Induction of remissions with vinblastine, actinomycin D, and bleomycin. Patients with disseminated testicular carcinoma were treated with the combination of vinblastine, actinomycin D, and bleomycin in an attempt to induce remission. Of 47 patients receiving an initial adequate trial of this regimen, 34% achieved a complete or partial remission; in the 18 patients with either no prior nonsurgical treatment or treatment with actinomycin D alone, the response rate was 61%. Those who attained complete response status enjoyed significant prolongation of life compared with the nonresponders or partial responders. Responses were seen in all histologic categories and were not related to the performance status of the patient at the start of the trial, to the total dose of drug in the first mouth of therapy, or the extent of hematologic toxicity produced by the drugs. Responders had a higher incidence of stomatitis than nonresponders.


Low dose bleomycin and methotrexate in cervical cancer. Twenty patients with recurrent and disseminated carcinoma of the cervix were treated with Bleomycin, 10 mg/m2 weekly and Methotrexate, 10 mg/m2 every fourth day. Twelve of the 20 (60%) had a greater than 50% shrinking of measured tumor masses lasting a median remission time of 7.5 months. The data suggest that combination protracted chemotherapy with these drugs of metastatic cervical cancer might improve the outlook of patients with this condition.


Effect of surgical and chemotherapeutic treatment on alpha-fetoprotein levels in patients with hepatocellular carcinoma. Quantitative determinations of serum alpha-fetoprotein (AFP) by radioimmunoassay in 193 patients with hepatocellular carcinoma have demonstrated a wide variation in serum levels that appear to be relatively constant for each patient by the time that diagnosis is made. If there is no therapeutic intervention the serum AFP usually follows a gradual increase as the tumor progresses. A few patients have a fall in serum AFP as a preterminal event. Various forms of chemotherapy cause only minor and transient decrease in serum AFP. Surgical resection of tumor produces an immediate fall that parallels the catabolic decay rate for AFP. All AFP-positive patients treated with surgery had recurrence of their tumor with a rise in serum AFP preceeding clinical discovery. The correlation of serum AFP and effective treatment is demonstration of the usefulness of this oncofetal protein marker as an indicator of neoplastic activity for hepatocellular carcinoma and tumors with embryonal cell components and possibly for some other entodermally derived neoplasms.


Fetal antigens in retinoblastoma.

Fetal antigens in retinoblastoma. Plasma carcinoembryonic antigen (CEA) and alpha fetoprotein (AFP) levels were studied in five patients with retinoblastoma. CEA and AFP were found elevated in four of five patients prior to therapy. Following enucleation and/or radiation therapy, the elevated values were significantly lowered. This preliminary study suggests that CEA and AFP levels may be of value in the diagnosis of retinoblastoma, and that serial CEA levels may be useful in evaluation of retinoblastoma patients after treatment.


Gonadal and extragonadal yolk sac carcinomas: a clinicopathologic study of 14 cases. Fourteen cases of yolk sac carcinoma, 10 occurring in gonadal, and four in extragonadal sites, seen at the Indiana University Hospitals from 1949 to 1974, were analyzed with respect to pathologic features, laboratory findings, and clinical course. Their histologic appearance was similar regardless of the site of origin. Two basic histologic types were observed--the more common endodermal sinus pattern and the rare polyvesicular vitelline form. The prognosis is unfavorable, but three of our cases exhibited objective responses to chemotherapy. In our small series, the better prognosis of testicular yolk sac carcinomas in children found by some authors was not evident. Four of the 6 patients with yolk sac carcinoma in which serum alpha-fetoprotein determinations were performed showed positive results. Three of these cases had residual or metastatic disease clinically. The demonstration of alpha-fetoprotein in the serum of patients with yolk sac carcinoma lends further support to the yolk sac origin of these tumors and could also prove to be of prognostic value by indicating the presence of residual or recurrent disease.


Alpha-1-fetoprotein in a patient with a poorly differentiated carcinoma of the anterior mediastinum. A case of a mediastinal tumor which was associated with the production of AFP is described. The diagnostic significance of an elevated serum concentration of AFP is reviewed in both neoplastic and non-neoplastic clinical situations.


Effective bone palliation as related to various treatment regimens. Various dose-time treatment plans have been used to obtain long duration pain relief in patients with metastatic bone disease. Very little has appeared in the literature evaluating the relationship of dose and fractionation to initial, delayed and permanent bone pain relief. At the Swedish Hospital Tumor Institute, 152 treatment fields in 110 patients were evaluated, with a clinical follow-up in many of over five years. Those treated at lower total doses with less fractionation achieved the same quality and duration of pain relief as higher doses. Treatment plans also were compared using the Ellis method of nominal standard dose.


C-type virus-associated antigens and their relevance to human leukemia control. This review describes oncogenic C-type virus and virus-associated antigens as phenotypic expressions of virus activities in malignant cells and cell surface antigens acquired by malignant transformation. The range of host-immune responses to these antigens in experimental systems is discussed in relation to the immunotherapy and immunoprophylaxis of human cancer patients.


Autogenous immunity to endogenous RNA tumor virus: humoral immune response to virus envelope antigens. Autogenous immune sera from several strains of mice have been examined for type-, group-, or interspecies-specific reactivities against leukemia virus envelope antigens and virus-induced cell surface proteins. The natural antibody of these test sera react with gp69/71, gp43, and p15 structural components on murine leukemia viruses including AKR, Friend, Rauscher, Moloney, and xenotropic BALB:virus-2. Furthermore, comparable radioimmune titration curves are obtained when these viruses are used in radioimmune precipitation assays. Competition experiments, however, suggest that natural immune sera are predominantly type specific and only weakly cross-react with the Rauscher or Friend virus. Natural immune sera react with the virion envelope but not with the virus-induced cell surface antigen. With respect to the biological activity of autogenous immune sera, there appears to be an inconsistency between the spectrum of virus-precipitating antibody and virus-neutralizing antibody. Although normal mouse serum readily neutralizes xenotropic viruses (BALB:virus-2), only weak neutralization of the ecotropic viruses can be achieved in vitro. Although there is a lack of direct evidence to indicate that autogenous immunity to murine leukemia virus is involved in the control of virus-mediated neoplasia, several empirical correlations point in this direction.


Cell-mediated immunity to leukemia virus- and tumor-associated antigens in mice. Cell-mediated immune reactions appear to play an important role in resistance against growth of leukemia cells in mice. Possible mechanisms for in vivo protection in two tumor systems are discussed. These tumor models, which are a Friend leukemia virus-induced transplantable tumor, FBL-3, and primary murine sarcoma virus (MSV) -induced tumors, are strongly antigenic; under some conditions, tumors regress completely. In mice with regressing FBL-3 tumors, cell-mediated cytotoxicity was measured by release of [125I]iododeoxyuridine. The response was biphasic, with an initial peak at 10 days and a 2nd peak after 30 days. A boost in reactivity could be elicited by later challenge with tumor cells. All of the reactivity was dependent on T-cells, being eliminated by treatment with anti-theta plus complement. The specificity of the reactions was not completely defined, but it was consistent with Friend type-specific antigen plus broader, common antigens. In mice with regressing MSV tumors, strong cell-mediated cytotoxicity, measured mainly by release of 51Cr, was seen against RBL-5, a Rauscher virus-induced leukemia. A single peak of response occurred at about 14 days after virus inoculation. Upon later challenge with RBL-5 cells, a vigorous and rapid secondary response was elicited, mainly in the region of tumor challenge. This cytotoxic reactivity and in vivo resistance to leukemia.lso was completely dependent on T-cells. In addition, macrophage-mediated inhibition of leukemia cell growth in vitro was seen in this system at the time of peak tumor development. The 51Cr release cytotoxicity was specific and directed primarily against an antigen, MEV-SA1, associated with mouse endogenous C-type viruses. The macrophage-induced growth inhibition appeared to be nonspecific. In both the FBL-3 and MSV tumor systems, protection against tumor growth could be adoptively transferred by immune lymphoid cells. In addition to induction of cell-mediated immunity by tumor cell or virus inoculation, cell-mediated cytotoxic reactivity was found to occur naturally in most young mice. This natural killer activity was quite distinct from the experimentally elicited reactions, being mediated by N-cells, a subpopulation of lymphoid cells with no clearly identifiable cell surface markers. The natural cytotoxicity was also directed against antigenic specificities different from those recognized by the MSV-immune cells. The central issue in all of these studies has been to determine the relationships between the in vitro-detected cell-mediated reactivity and in vivo resistance to leukemia.


Immune response to leukemia virus and tumor-associated antigens in cats. Cats represent an unusually valuable model for studying the role of the immune response to leukemia, lymphoma, and other mesodermal neoplasms. The agents that cause spontaneous feline leukemias, lymphomas, and fibrosarcomas, the feline leukemia and sarcoma viruses, are well characterized. A specific tumor cell membrane antigen, designated the feline oncornavirus-associated cell membrane antigen (FOCMA) has also been described. Feline leukemia and feline sarcoma viruses are antigenically indistinguishable, and FOCMA is common for both. Both laboratory-induced and spontaneous feline leukemias, lymphomas, and fibrosarcomas are available for study. A clear correlation has been shown between the resistance of cats to development of lethal tumors following inoculation of feline sarcoma virus and the presence of high humoral antibody titers to FOCMA. The geometric mean antibody titer to FOCMA for cats that resisted growth of fibrosarcomas was more than 20-fold higher than the mean for cats that succumbed to lethally progressing tumors. Cats with induced or spontaneous leukemia or lymphoma also have either no detectable FOCMA antibody or very low levels. Conversely, some cats resist development of leukemia or lymphoma following natural exposure to feline leukemia virus in leukemia cluster households, and these cats have high FOCMA antibody titers. These results support the concept of a natural immunosurveillance mechanism against leukemia or lymphoma development in an outbred mammalian species.


In vivo and in vitro measurements of the relationship of human squamous carcinomas to herpes simplex virus tumor-associated antigens. An additional 244 unfiltered sera have now been studied in a series of controlled, coded tests to determine the relationship of squamous carcinomas of the head and neck and cervix to the presence of complement-fixing antibodies to herpesvirus-tumor-associated antigens (HSV-TAA) in both tumor-bearing and cured patients. Ninety % of sera from patients with squamous carcinomas had antibodies to HSV-TAA, in contrast to 11% of sera from patients with nonsquamous cancers and 4% of sera from noraml individuals. The temporal relationship of Stage 1 laryngeal carcinomas suggests that HSV-TAA appearance precedes the immune defects. An in vitro correlate of the previously demonstrated specific delayed hypersensitivity reactions in controlled skin tests of squamous carcinoma patients with HSV-TAA is reported. In leukocyte migration inhibition tests, the migration indices after incubation with HSV-TAA of peripheral blood leukocytes from patients with squamous carcinoma (x = 0.847) were in definite contrast to migration indices seen for normal leukocytes (x = 1.037) and patients with nonsquamous solid cancers (x = 1.03). Thus, these polypeptides elicit both humoral antibody response and cell-mediated reactivity.


Serological and epidemiological considerations of the role of herpes simplex virus type 2 in cervical cancer. To assess the possible biological significance of the observations that women with cervical cancer tend to be younger at first intercourse than control women, data from 1823 women were analyzed for the relationship between age at 1st intercourse and number of sex partners. Women who were younger at first intercourse had more sex partners than did women who were older at first intercourse. The interdependence of age at first intercourse and number of sex partners does not exlude the possibility that intercourse at an early age represents a biologically significant event in which the neoplastic process is initiated. However, it is equally possible that younger women at first intercourse may have multiple sex partners and be at greater risk of coming in contact with a putative oncogenic agent later in life. In addition, sera from patients with herpesvirus infections were assayed for cross-reacting and type-specific antibodies. Approximately 80% of the total antibody activity was to the cross-reacting antigen and only 20% was to the type-specific antigens in the sera of patients infected with either type 1 or type 2 virus. Among patients infected with both types of virus, less antibody activity to the type-specific antigens and more antibody activity to the cross-reacting antigens were found. These observations are discussed with respect to case-control seroepidemiological studies.