Blood lead levels in sheep exposed to automotive emissions. The lead content of whole blood of 62 sheep grazed continuously for 6 months near a major highway was compared with 38 sheep from a nearby uncontaminated area. Mean values of 0.90 and 0.20 microgram/ml were obtained. Four sheep from the contaminated area were placed in an uncontaminated paddock and the lead content of the blood decreased rapidly during the first 10 days and thereafter more slowly. After 185 days, blood lead levels had still not quite reached normal levels. Animal from an uncontaminated area showed an immediate rise in the blood lead levels when placed near a major highway. Sheep placed in a concrete pen away from motor vehicle exhaust emissions showed a rise in lead levels when fed with forage cut from the verges of a busy highway. Sheep placed near a highway and fed with forage from an uncontaminated area showed an increase of lead levels in the blood, comparable to that of the previous experiment. It was concluded that lead uptake by ingestion of polluted forage and by inhalation of airborne particulates are both mechanisms responsible for enhanced lead levels in ovine blood.


A screening technique for assessing effects of pesticides on population and activities of non-target soil microorganisms. A simplified technique for assessing the effect of pesticides on non-target soil microorganisms was developed. Changes in microbial population determined by the dilution plate method, in nitrification by the soil perfusion technique, and in oxygen consumption using a differential respirometer are the only measurements required. A comparison of the effects observed for three pesticides, dieldrin, chlorpyrifos and Vorlex, in three soil types with those produced by an antibiotic, a fungicide, a nitrification inhibitor and steam pasteurization of the soil clearly demonstrate the effectiveness of the simple technique.


The value of pregnancy-associated alpha2-glycoprotein in patients with colorectal cancer. Serum pregnancy-associated alpha2-glycoprotein (alpha2PAG) levels have been measured in 67 patients with colorectal cancer. Postoperative changes in alpha2PAG concentrations were observed in patients undergoing apparently curative surgery and compared with results in patients with residual local or metastatic tumour. Significant increase in alpha2PAG levels were found only in those patients with residual or disseminated tumour. In this study alpha2PAG levels closely paralleled the clinical course of the disease in many patients.


Alpha-fetoprotein concentrations measured by radioimmunoassay in diagnosing and excluding hepatocellular carcinoma. Serum alpha-fetoprotein (AFP) concentrations were estimated by sensitive radioimmunoassay in 30 patients with cirrhosis complicated by hepatocellular carcinoma and in 100 patients with cirrhosis in whom malignancy was excluded. Twenty-nine of the 30 patients with hepatocellular carcinoma had concentrations above 10 IU/ml (10.5 ng/ml) (median 3500 IU/ml (3675 ng/ml)), whereas only one of the 100 patients with cirrhosis and no tumour development had a raised concentration. Eleven out of 20 patients in whom hepatocellular carcinoma had developed in an apparently normal liver had raised AFP concentrations. In this group the differential diagnosis is usually secondary carcinoma, and three of 50 such patients had AFP concentrations above 10 IU/ml. Noting raised AFP concentrations is thus of considerable value both in detecting and in excluding hepatocellular carcinoma in cirrhosis, for in this case such concentrations gave only 1% false-positive and 3% false-negative results. They are less useful, however, in distinguishing between primary tumours arising in patients without cirrhosis and secondary hepatic deposits, giving 6% false-positive and 45% false-negative results.


Distribution of neurons projecting to the retina of Caiman crocodilus. Horseradish peroxidase (HRP) was injected into the vitreous of the eye, the orbital cavity, or the optic tectum of Caiman crocodilus. Following intravitreal injections, retrograde transport of the enzyme was observed bilaterally, but predominantly contralaterally, in a large oblong field of cells at the isthmic level of the midbrain, bounded medially by the trochlear nucleus and laterally by the nucleus isthmi. Control injections of HRP into the orbital cavity and eye muscles labelled motoneurons of the extraocular muscles but not cells of this isthmic field. The field is therefore the source of a projection efferent to the retina in Caiman. Injections of HRP into the dorsal optic tectum produced a fine pattern of anterograde labelling in fibers projecting to the region in which retinopetal cells were identified, but no retrograde labelling of cells in this area. In contrast, the nucleus isthmi showed dense anterograde labelling of fibers and terminals as well as retrograde labelling of cells following tectal injections. These results extend recent evidence for a close evolutionary relationship between the order Crocodilia and modern birds, since birds also possess a well-developed retinopetal system derived from a cell group in the isthmic midbrain.


Uptake and retrograde axonal transport of various exogenous macromolecules in normal and crushed hypoglossal nerves. Macromolecular tracers were injected into the tongue or around a crush in mouse hypoglossal nerves. At various times thereafter, the tracers were histochemically localized on the basis of peroxidase activity. The distribution of reaction product was then examined using light microscopy in order to study the influence of molecular charge and size on uptake and retrograde axonal transport from the periphery or from the crushed axon. Of various proteins with peroxidase activity, horseradish peroxidase and cytochrome-c showed the greatest penetration into axons proximal to the crush. Following injection into the tongue, intra-axonal cytochrome-c was detectable in some of the peripheral branches but not any of the other proteins. Retrograde transport to the nerve cell bodies was demonstrated for horseradish peroxidase and cytochrome-c, both from the tongue and from the axonal crush but not for microperoxidase, myoglobin, hemoglobin, lactoperoxidase and catalase. The number of neuronal cell bodies having detectable reaction product was higher for peroxidase-injected than for cytochrome-c-injected animals. Ferritin and iron-dextran (Imferon) also accumulated in hypoglossal neurons, but this could be detected only after repeated injections into the tongue. Uptake and retrograde transport from the tongue or from the crush occurred both for anionic and for cationic horseradish peroxidase. This is interpreted as evidence against absolute specificity in the uptake and transport of macromolecules on the basis of electrical charge.


CSF electrophoresis: an adaptation using cellulose acetate for the identification of oligoclonal banding. An adaptation of cellulose acetate electrophoresis for studying concentrated cerebrospinal fluid is described. Two hundred and twenty-one patients have been studied, and the specificity for multiple sclerosis and sub-acute sclerosing panencephalitis is discussed. This has been positive for oligoclonal banding (OB) in 79% of patients with clinically definite multiple sclerosis.


Effectiveness of local radiotherapy in colorectal carcinoma. The records of 144 patients with colorectal carcinoma treated with radiotherapy were reviewed. Ninety-two patients received treatment for palliative purposes only (bleeding, pain, mucus discharge, tenesmus, mass). Good palliation was achieved in the majority of patients with a dose on the range of 1200 to 1400 rets. Thirty-seven patients received radical doses of radiation for recurrent or inoperable cancer. Five out of 37 (14%) are alive without evidence of disease, 24 to 84 months following radiotherapy. Fifteen patients received pre- or postoperative radiotherapy as an adjuvant to surgery. Six out of the 15 (40%) are alive without evidence of disease.


Radiation treatment of primary carcinoma of the female urethra. Because of the rarity of the primary carcinoma of the female urethra, there has been limited experience of individual institutions in management of this disease. Treatment by radiation alone required radium implant skill in early cases and individualized consideration for integrating external and internal irradiation in more advanced cases. From 1961 to 1975 a total of 16 women with histologically verified primary urethral carcinoma were given radiation treatment at the University of Maryland Hospital. Six of 16 patients were primarily treated for palliation only and 10 were receiving curative radiation treatment. The results show an excellent local control in anterior urethral disease (3/3), entire urethral disease (2/3) and urethral disease with involvement of vulva and/or vagina (3/3). Bladder neck, parametrial, inguinal lymph node, or paraortic involvement represents a poor prognosis and failure is close to 100%. Eight of 10 patients who were treated with aggressive radium alone or combination of external irradiation and interstitial therapy show complete control of disease (80%). Total dosage between 5500 to 6500 rad appears to be adequate to control the disease. Palliative treatment by irradiation alone can only offer a short term symptom-free result, and should be considered as an occasional alternative modality of choice.


Leiomyosarcoma of the small and large bowel. From 1950 through 1974, a total of 108 cases of primary intestinal leiomyosarcoma were seen at the Mayo Clinic. Most of these uncommon tumors occurred in the fifth and sixth decades of life, and more of them in men than in women (2.6:1). There were 73% in the small bowel, 25% in the large bowel, and 2% in the anus. Gastrointestinal bleeding and pain were the two most common signs at presentation, and they led to surgical exploration in all cases where they appeared. By the time surgery was performed, only 48% of the tumors could be resected with hope of cure. Within that group of cases, 50% of the patients survived 5 years, but only 35% survived 10 years, late recurrence being common. The histologic grade of the tumor affected survival to erroneous early optimism in prognosis.


Specificity of response in hamster cells induced to produce plasminogen activator by the tumor promoter, 12-O-tetradecanoylphorbol-13-acetate. A phorbol ester promoting agent, 12-O-tetradecanoylphorbol-13-acetate, enhances plasminogen activator production in hamster cell lines that synthesize plasminogen activator immunologically identical with plasminogen activator of normal hamster lung cells. 12-O-Tetradecanoylphorbol-13-acetate exhibits a high degree of specificity in the type of plasminogen activator evoked, which is always of the lung form. No enhancement of plasminogen activator production was detected in hamster cell lines synthesizing plasminogen activator of different antigenic form, although in the presence of 12-O-tetradecanoylphorbol-13-acetate, cells of one such line initiated synthesis of plasminogen activator of the lung form.


Enhancement of DNA chain breakage by bleomycin A2 in the presence of microsomes and reduced nicotinamide adenine dinucleotide phosphate. Chain breakage in DNA induced by bleomycin A2 (BLM) was enhanced more than 150-fold by the reduced nicotinamide adenine dinucleotide phosphate-dependent electron transport system of rat liver microsomes. However, the enhancement effect on DNA was partially reduced by the preincubation of BLM with the microsomal systems. BLM-Cu2+ was found to stimulate considerably microsomal reduced nicotinamide adenine dinucleotide phosphate-dependent oxygen consumption and malondialdehyde formation, whereas BLM inhibited both of the effects. These findings suggest that the pharmacological action of BLM is strongly affected by a membrane system, such as microsomes, that produces free radicals.


Dihydrotestosterone concentration in prostate cancer tissue as a predictor of tumor differentiation and hormonal dependency. Tissue dihydrotestosterone and 5alpha-reductase (delta4-3-ketosteroid-5alpha-oxidoreductase) levels have been measured in prostates of patients with cancer and benign prostatic hypertrophy; significant decreases in average values for both of these biochemical parameters were noted in prostate cancer compared to benign prostatic hypertrophy, although individual values overlapped in both groups. Prostate cancer tissue dihydrotestosterone levels appeared to correlate better than did either histological tumor grading or 5alpha-reductase with the ultimate clinical response to antiandrogen therapy. These results suggest that assay of tissue dihydrotestosterone levels in prostate cancer should be further explored as a possible marker for tumor differentiation.


Feline sarcoma virus in vitro infection of human cells. Influence of chemical carcinogens on focus formation. Human fibroblast cells treated with benzo[alpha]pyrene (BP), aflatoxin B1 (AFB1) or N-acetoxy-2-fluorenylacetamide (A-AAF) inhibited Snyder-Theilen feline sarcoma virus (ST-FeSV) focus formation. Inhibition of focus formation resulting from chemical treatment was not related to cytotoxic concentrations of chemicals in that little or not effect on cells surviving treatment was observed. Maximum inhibition of focus formation occurred with BP when the cells were treated before infection. By contrast, maximum inhibition of focus formation occurred with A-AAF and AFB1 when the cells were treated after virus infection. Inhibition of focus formation by BP and AFB1 was eliminated when virus infected cells were treated 48-96 h post-infection. While no infectious virus was detected in either chemical treated or untreated ST-FeSV virus infected cultures, comparable levels of virus-directed RNA dependent DNA polymerase enzyme assay (RDDP) activity were found in both treated and untreated cultures. The data show that the inhibitory effect on focus formation is chemically mediated while the inhibition of virus synthesis is not.


Gut endocrinology: correlation of physiology and morphology. The gastrointestinal tract is a large and complex endocrine organ. Elucidation of its nature, and the actions of its individual hormonal components, has advanced in recent years following the purification and sequencing of six probable hormones and the subsequent development of radioimmunoassay techniques for their measurement and immunocytochemistry for the determination of their cellular origin. Immunoassay and immunocytochemistry are clearly complimentary to each other in providing a fuller understanding of the dynamics of endocrine function at the cellular level. This is illustrated in the examples described in this paper.


Serum steroids in normal males and patients with prostatic diseases. The subjects investigated comprised 57 normal males between 30 and 80 years of age; 40 patients between 50 and 80 years of age suffering from benign prostatic hypertrophy (BPH), and 11 untreated prostatic carcinoma (Ca) patients aged between 57 and 79 years. Serum concentrations of oestradiol, pregnenolone, progesterone, 17alpha-hydroxyprogesterone, androstenedione, testosterone, 5alpha-dihydrotestosterone and androsterone were determined from a single serum sample (1.6 ml). Oestradiol was determined by an automated non-chromatographic radioimmunoassay, while other steroids were determined by radioimmunoassays, after solvent extraction and chromatographic purification on Lipidex-5000TM microcolumns. When patient groups were compared with the 25 normal males between 50 and 80 years of age, several conclusions could be drawn. Serum concentrations of 5alpha-dihydrotestosterone (P less than 0.01) and 17alpha-hydroxyprogesterone (P less than 0.001) were both significantly higher in the BPH patients when compared with the normal males. This trend was also apparent in the serum concentrations of progesterone and testosterone in the older BPH patients. Although the mean concentrations of 5alpha-dihydrotestosterone and 17alpha-hydroxyprogesterone were slightly higher in carcinoma patients than normal males, these differences were not statistically significant. No differences were seen in the concentrations of pregnenolone, androstenedione, androsterone and oestradiol between normal subjects and patients with BPH or prostatic Ca.


Sex determination in nuclei of amnion fluid cells. Hypotonic treatment of amnion fluid cell-nuclei was found to enhance demonstration of Y chromatin and of autosomal fluorescence in interphase cell-nuclei following quinacrine mustard staining. In contrast, visualization of the Barr body was not improved by this treatment. Nuclear sex determination on cellular material from 200 samples of amniotic fluid was unequivocal in 197 cases; in only 3 cases were the findings uncertain. There were no erroneous determinations. In one instance maternal cellular tissue was observed in smear preparations. Nuclear sex determination presents a simple method of supplementing and substantiating prenatal chromosomal diagnosis.


The effect of mercury derivatives of fluorescencein on allotransplant kidney in experiment. Mercurascan (MSC), a mercury derivative of fluorescein, was studied as to its effect on the survival of allogeneic kidney grafts in dogs. A single perfusion of the donor's kidney with saline solution with MSC added in a ratio of 1 mg/100 ml was found to have significantly extended the graft survival time from 10.8 days (SE +/- 0.6) to 15.4 days (SE +/- 1.6). Graft survival time extension proved to be even more significant after a single dose of MSC given to the donor (0.5 mg/kg b.w.) and repeated administration of an equal MSC dose to the recipient twice weekly. Listed are detailed morphological changes in tissue specimens from the kidney, lymph nodes, spleen, and many other organs of a dog surviving for 42 days. A very moderate rejection lesion with merely tiny lymphocytic infiltrates was proved in the kidney. Lymph node structure was almost entirely obliterated with lymphoid tissue extinction. There was loss of white pulp in the spleen. The absence of morphological changes in the other organs suggested good tolerance of the preparation. More studies are called for to elucidate the complex MSC intervention in the process of rejection.


Influence of food on the bioavailability of drugs. Food intake exerts a complex influence on the bioavailability of drugs. It may interfere not only with tablet disintegration, drug dissolution and drug transit through the gastrointestinal tract, but may also affect the metabolic transformation of drugs in the gastrointestinal wall and in the liver. Different food components can have different effects, and food may interact in opposite ways, even with drugs that are chemically related. Therefore, the net effect of food on drug bioavailability can be predicted only by direct clinical studies of the drug in question. As judged mainly from single meal, single dose studies, food intake enhances the bioavailability of several different drugs, such as propranolol, metoprolol, hydrallazine, hydrochlorothiazide, canrenone (from spironolactone), nitrofurantoin, erythromycin (stearate), dicoumarol, phenytoin and carbamazepine, but reduces that of drugs such as isoniazid, rifampicin, tetracycline, penicillin and ampicillin, while having no consistent effect on the bioavailability of metronidazole, oxazepam, melperone, propylthiouracil, sulphasomidine and sulphonylureas. For some drugs such as digoxin and paracetamol, the rate but not the extent of absorption is reduced. Food may enhance bioavailability even though, or rather because, the rate of gastric emptying is reduced; this is apparently the case with hydrochlorothiazide and nitrofurantoin. The food induced enhancement of bioavailability of propranolol, metoprolol and hydrallazine is probably due to reduced first pass metabolism of these drugs, while food induced improvement of drug dissolution may explain the enhanced bioavailability of carbamazepine, canrenone, dicoumarol and phenytoin. An increased gastrointestinal pH may be in part the cause of the food induced reduction of the bioavailability of drugs such as isoniazid and tetracycline. In addition to single meal effects, repeated intake of protein-rich meals enhance, while carbohydrate-rich meals reduce, the rate of oxidation of antipyrine and theophylline. Moreover, intake of charcoal broiled meat markedly accelerates the oxidation of phenacetin and variably accelerates elimination of theophylline. Thus, food and its components and contaminants may have both short and long term effects on both the absorptive and biotransformation processes influencing systemic availability of drugs.


[Rubella in pregnancy: experimental studies on the value of gamma-globulin after rubella wild virus infection (author's transl)]. Rubella infection was produced with a wild rubella virus in 24 volunteers by intranasal application. Some of the subjects were given gamma-globulin intramuscular 24 hours later. In the control group clinically manifest rubella with massive viral excretion in the throat occurred in all in whom infection had taken place. In the majority of subjects rubella virus could be demonstrated in blood shortly before the skin rash appeared. High antibody titres occurred already 20 days after the infection. In subjects who were given gamma-globulin there were no clinical signs of rubella, viral excretion in the throat was diminished and shortened and viraemia was never demonstrated, antibody response was delayed and lower. But there was no significant decrease in the infection rate after gamma-globluin administration. It is concluded that early administration of a sufficient amount of rubella antibodies achieves inhibition of rubella virus multiplication and thus decreases significantly the risk of damage to the fetus.


[Radioimmunoassay of beta2-microglobulin in monoclonal gammopathies (author's transl)]. Serum concentration of beta2m were measured by radioimmunoassay in 78 healthy subjects and 80 patients with monoclonal gammopathies. The beta2m levels of normal sera were normally distributed with a mean concentration of 1610 microgram/l. 77% of values were between 1200 and 2000 microgram/l. The serum beta2m levels of patients with monoclonal gammopathies were significantly higher than those of the healthy subjects, but there was no significant difference when comparing the Ig class or the light-chain type. Urinary beta2m excretion of patients with monoclonal gammopathies (3--5400 microgram/l) were slightly higher than normal (15--113 microgram/l). Serum beta2m levels did not correlate with the serum levels of monoclonal immunoglobulins. This supports the hypothesis of the mutual independence of beta2m and Ig production.


[Changes in the activity of histamine-liberating enzymes in acute experimental anaphylaxis]. The authors examined the changes in the histamine level, DAO (diaminoxidase) activity and N--MT (N-methyltransferase) activity in blood (plasma) and tissues of lung and liver (N--MT was examined only in the indicated tissues) in 105 guinea pigs, 45 of whom were healthy and 60 sensibilised and examined during the anaphylactic shock. It was established that during the acute anaphylactic shock there was demonstrative hyperhistanemia and hyperhistaminasemia, which were not observed during experimental anaphylactic shock with subacute course. The activity of lung and liver N--MT in animals with acute and subacute anaphylactic shock did not differ from that of the healthy guinea pigs. The complete inhibition of DAO activity in healthy animals and those with anaphylactic shock did not induce change in the activity of N--MT as well. The latter did not alter after sharp or high level or moderate elevation of the tissue histamine for several days. This histamine was a specific substrate for this enzyme. N--MT is an inductive enzyme, which apparently does not participate in histamine metabolism during acute and subacute anaphylaxis in guinea pigs. For its change, probably, is necessary longer time.


[Audiovisual recordings of electromyographic examinations (author's transl)]. The presented method is to be an aid in the recording and documentation of single EMG examination results by the possibility of adding acoustic and clinical details to the previous method which was purely an optical method. This recording method is especially good for continuing education courses and for detailed storing of certain results, because the method may be reprodused indefinately. The method is not suitable for conventional documentation because 1) it is too costly and 2) it does not fulfill the qualitative requirements of a documentation.


Carcinogens and dicumarol: opposite effects on rat liver NAD(P)H dehydrogenation. The carcinogens, N-acetyl-aminofluorene, 7,12-dimethylbenzanthracene, 3,4-benzpyrene and 3-methylcholanthrene, increase the activity of the soluble enzyme D-T-diaphorase. This action is observed 24 h after the administration of these chemicals to rats. Dicumarol blocks this effect. Dicumarol does not inhibit the increase in activity of the microsomal aryl hydrocarbon hydroxylase system as elicited by 3,4-benz(a)pyrene and 3-methylcholanthrene. The functional significance of these findings and the possible role of cytosolic enzymic changes in chemical toxicity are discussed.


The effects of extracellular potassium and several drugs on the premature action potential and postextrasystolic potentiation. To investigate the mechanism of postextrasystolic potentiation of contraction (PESP), the effects of extracellular K concentration, isoproterenol, verapamil and caffeine were examined in canine ventricular muscle. The muscles were driven at 1 Hz, then a premature stimulus was applied to elicit a premature action potential and after a compensatory pause postextrasystolic stimulus was applied to induce PESP. At 5.9 mM K the premature action potential had a longer plateau than the preceding control action potential. The prolongation was dependent on the proximity, i.e. the interval between the premature stimulus and the preceding control action potential; the maximal enhancement of about 25% was obtained with a proximity of 70 ms. In parallel with this prolongation the post-extrasystolic contraction was increased more than 5-fold. However, PESP was obtained without prolongation of the premature action potential when [K]0 was reduced to 1.2 mM. Similarly, PESP under the effect of 10(-6) g/ml isoproterenol or 10(-5) M verapamil was not accompanied by prolongation of the premature action potential. Thus it is difficult to explain the PESP solely by the increased Ca influx during the premature action potential. Since caffeine abolished PESP, some other effects of the premature beat on sarcoplasmic reticulum might be responsible for the PESP.


Mechanisms of tachyarrhythmias, past and present. Several hypotheses, developed in the early years of arrhythmology, and frequently used for the explanation of many types of clinical tachyarrhythmias, are often based on experiments on unusual models, unphysiologic interventions, or, in some instances, on erroneously interpreted histologic findings. However, it can be demonstrated that several of these hypotheses are valid. Focal activity, unidirectional block, extremely slow conduction velocity, differences in conduction velocity in closely adjacent regions, reentry of the excitation wave, circulating excitation waves in small or large regions of the heart and other postulated mechanisms as, for example, entrance and exit block, local ventricular fibrillation, appear to be involved in the causative mechanisms of clinical tachyarrhythmias. A multicausal genesis of at least some of these must be considered seriously, particularly those which occur in acute or chronic coronary heart disease. A recent suggestion that the injury current might be involved in the genesis of ventricular tachyarrhythmias in acute myocardial ischemia and infarction is supported by observations on several properties of this current. Progress has been made in recent years but large gaps in our knowledge of mechanisms causing arrhythmias are still present.


Monoclonal xenogeneic antibodies to murine cell surface antigens: identification of novel leukocyte differentiation antigens. Hybrid myeloma cell lines secreting monoclonal antibodies to mouse cell surface antigens have been prepared. Spleen cells from a DA rat immunized with B10 mouse spleen cells that had been enriched for T cells were fused to cells from a nonsecreting mouse myeloma line (NSI). The presence in the culture supernatants of antibodies binding to mouse spleen cells was tested by a binding assay with 125I-labeled anti-rat IgG. From a large number of positive cultures, ten independent hybrid clones were purified, each secreting a different antibody. Each antigenic target was analyzed by (a) gel electrophoresis of immunoprecipitated 125 I-labeled cell surface molecules, (b) heat stability, (c) strain and species distribution and (d) cross-inhibition of binding of different monoclonal antibodies. It was concluded that the ten monoclonal antibodies regognized four types of antigen. One was the heterophile, heat-stable, Forssman antigen. The second (mol.wt. 210 000) appears to be a major 125I-labeled lymphoid cell surface protein. The third, a minor component of spleen cells, was precipitated as two polypeptides of mol.wt. 190 000 and 105 000. Five IgG-secreting clones identify the fourth antigen, a heat-stable, possibly glycolipid component expressed on mouse red blood cells and also on thymocytes. Cross-inhibition studies suggest that these last monoclonal antibodies bind to overlapping, but not identical, determinants. The class and chain composition of the monoclonal antibodies were studied by gel electrophoresis, isoelectric focusing and ability to lyse red blood cells and thymocytes.


ATP in cholinergic nerves:evidence for the axonal transport of a stable pool. The ATP content of the cholinergic electromotor nerves of Torpedo marmorata has been measured. After freezing and thawing of pieces of nerve, their ATP content dropped rapidly to about 5% of that of pieces extracted immediately. The amount of ATP in nerves incubated in Torpedo Ringer remains at a high level over a long period. In nerves ligated for seven days there is no accumulation of total ATP above the ligature. If, however, the ligated nerves are frozen and thawed for 20 min before extraction, the residual ATP shows significant proximal accumulation at the ligature. This transport is not due to the accumulation of mitochondria at the ligature.Acetylcholine, measured in the same nerve extracts of nerve segments, is also axonally transported. The ATP which is resistant to hydrolysis is probably sequestered within a subcellular organelle. The possibility that its accumulation at ligatures may indicate movement of cholinergic synaptic vesicles which are known to contain ATP and acetylcholine is considered.


Alpha benzene hexachloride inhibition of aflatoxin B1-induced hepatocellular carcinoma. A preliminary report. Alpha benzene hexachloride protected against the development of liver carcinoma in male albino Fisher rats ingesting aflatoxin B1.


Binding bovine immunoglobulins to anterior lobe cells of the hypophysis. Cells of the anterior lobe of the bovine hypophysis, show an affinity for immunoglobulins IgG, IgA and IgM. The phenomenon either provides a message to stimulate cellular secretion or to excite mechanisms associated with the synthesis and release of Ig's or their determinants.


The kinetics of ion movements in the gramicidin channel. The main features of the ion permeability of gramicidin channels are summarized. The significance of maximums in the single channel conductance-concentration curves, of concentration-dependent permeability ratios, and or current-voltage curves with concentration-dependent form, as well as of other features, is discussed in terms of the mechanism of the ion transfer processes. The observations are then shown to be accounted for by rate theory expressions derived for a model pore consisting of two sites in series and in which ions are not permitted to pass each other. The status of other models is briefly reviewed.


Ion channels formed by chemical analogs of gramicidin A. Channel-forming peptides such as gramicidin A offer the opportunity to study the relationship between chemical structure and transport properties of an ion channel. This article summarizes a number of recent experiments with chemical analogs and derivatives of gramicidin A using artificial lipid bilayer membranes. The introduction of negative charges near the channel mouth leads to an increase in the cation transport rate. Hybrid channels consisting of a neutral and a negatively charged or of a positively and a negatively charged half-channel may be formed. The current-voltage characteristic of these hybrid channels exhibits a pronounced asymmetry. Experiments with charged derivatives of gramicidin A have been used in order to distinguish between different structural models of the dimeric channel; these studies strongly support Urry's model of a single-stranded, head-to-head associated helical dimer. In a further set of experiments gramicidin analogs with modified amino acid sequence were studied. It was found that a single substitution (tryptophan replaced by phenylalanine) leads to marked changes in the conductance of the channel. Analogs with a simplified amino acid sequence such as (L-Trp-D-Leu)7-L-Trp or L-Trp-Gly-(L-Trp-D-Leu)6-L-Trp are able to form cation permeable channels with similar properties as gramicidin A.


Identification of A1 macroglobulin and A2 macroglobulin in rat serum by a two-dimensional quantitative immunoelectrophoresis with intermediate gel. A1 macroglobulin was identified in rat serum by a two-dimensional quantitative immunoelectrophoresis with intermediate gel containing monovalent anti-A1M rat serum. A second serum macroglobulin A2M antigenically related to A1M was identified in turpentine-stimulated or irradiated rats.


Immunological studies with the HA1 and HA2 polypeptides of influenza A virus haemagglutinin. HA1 and HA2 polypeptides of influenza A virus haemagglutinin (HA) were separated in purified form using electrophoresis in SDS containing polyacrylamide gels (PAGE) or chloroform-methanol extraction. The populations of HA1 polypeptides were immunogenic but considerably less so than the intact HA molecule and induced antibody which cross-reacted with influenza A and B viruses. After absorption with heterologous influenza B virus, the cross-reacting antibodies were removed and the HA1 antisera then possessed antibodies which reacted only with the cross-reactive (CR) determinants of the HA of the homologous influenza A virus and viruses of the same subtype. Neither strain-specific (SS) nor virus-neutralizing antibodies were detected in these anti-HA1 sera. HA2 polypeptides were less immunogenic and anti-HA2 antisera after absorption with influenza B virus failed to react with influenza A virus in immuno double diffusion tests and only reacted with partially denatured HA in the more sensitive single radial diffusion tests.


Carcinosarcomas and mixed mesodermal tumors of the ovary: a clinicopathologic study of six cases. A clinicopathologic study of three ovarian carcinosarcomas and three mixed mesodermal tumors of the ovary is presented. None of the patients survived for more than 1 year, irrespective of chemotherapeutic treatment or the extent of surgery. No prognostic differences were noted between the histologic patterns of varying tumor sizes of carcinosarcomas and mixed mesodermal tumors. The association of these two entities with other neoplasms of Müllerian nature suggests a multicentric tumor response of Müllerian tract derivatives. The necessity of adequate sampling, both for correct diagnosis and to exclude the possibility of teratoma (which behaves in a more benign fashion), is emphasized.


A comparison of the short-term effects of ibuprofen and diclofenac in spondylosis. In this controlled, single-blind parallel group study, the effect of ibuprofen 1200 mg daily was compared with diclofenac 75 mg daily. Thirty patients entered the study, randomized into two groups, each group receiving one tablet three times daily for two weeks. A one-week wash-out period (i.e. a drug-free period during which only physiotherapy was given), preceded and followed the treatment on active drug. Assessments were made by the same clinician throughout who was unaware of the treatment of individual patients. Statistically significant improvement was shown by patients receiving ibuprofen for the degree of pain relief at rest and improvement was also shown for the degree of pain at rest and during exercise, for pain relief during exercise and for spinal flexion. Patients who received diclofenac showed statistically significant improvement for forward flexion, together with improvement for the degree of pain on exercise. Side-effects were very few.


Analgesic efficacy and acceptability of fenoprofen combined with paracetamol and compared with dihydrocodeine tartrate in general practice. A single-blind between-patient study in general practice was made to examine the acceptability and analgesic efficacy of a combination of fenoprofen (200 mg) and paracetamol (500 mg) given 3 to 4 times daily for a period of 4 to 14 days in the treatment of acute and chronic pain. The combination was compared with dihydrocodeine tartrate (30 mg) in two groups of 75 patients suffering from a wide variety of presenting conditions, including osteo-arthritis, spondylitis, lumbago, sciatica, non-articular rheumatism and trauma. Clinical and patient assessment indicated that the combination was an effective analgesic combination, significantly better in success rate than dihydrocodeine tartrate and with a much lower incidence of side-effects (13% of patients receiving fenoprofen and paracetamol reported side-effects compared to 37% for the dihydrocodeine group). The combination demonstrated particular success in the treatment of non-articular rheumatism.


The acute effect of insulin on renal haemodynamics and protein excretion in diabetics. The effect of IV injection of 7 to 8 I. U. of insulin on renal haemodynamics and on urinary excretion of beta-2-microglobulin and of albumin was examined in 5 juvenile diabetics. Plasma glucose decreased from a mean value of 250 mg/100 ml to 117 mg/100 ml during the first 85 min after insulin. None of the patients had symptoms of hypoglycaemia and plasma adrenaline did not increase. There was no change in arterial blood pressure after insulin whereas pulse rate increased from 66/min to a maximum of 75/min. Glomerular filtration rate (GFR) and renal plasma flow (RPF) were decreased by 9 per cent and 13 per cent, respectively, during the first 90 min after insulin (2 p less than 0.01). There was also a statistically significant decrease in urine flow and urine secretion of several electrolytes, while filtration fraction remained almost constant. IV insulin decreased urinary excretion of beta-2-microglobulin and increased albumin excretion (2 p less than 0.05). The albumin excretion induced by insulin is most likely due to increased amounts of filtered albumin, the mechanism of which remains unexplained.


Early idiopathic hemochromatosis with absent stainable bone marrow iron stores. Early idiopathic hemochromatosis was diagnosed in a 52-year-old woman during the course of a hospitalization for another disorder. Quantitative studies revealed massive hepatic parenchymal siderosis and absent stainable marrow iron. Additionally, intestinal absorption and hepatic sequestration of radiolabeled iron were elevated. These studies in a patient with very early hemochromatosis, before organ damage, complement observations previously made in patients with advanced disease and provide further support for the concept that this disorder may result from impaired reticuloendothelial handling of iron.


Sequential staining of DNA-aldehyde with Schiff's reagent and acriflavine-SO2. Acid hydrolysed DNA of rat liver was stained with Schiff's reagent at pHs 1.7 or 3.0 followed by staining with acriflavine-SO2 at pH 2.0 as well as with acriflavine-SO2 followed by Schiff's reagent at pH 1.7 or 3.0. Nuclei stained with Schiff's reagent at pH 1.7 were brown-yellow and an analysis of their absorption characteristics revealed two peaks--one at 470 nm and the other at 570 nm. Although nuclei stained with Schiff's reagent at pH 3.0 followed by acriflavine-SO2 were deep magenta in colour, they also showed similar peaks of maximum absorption. Identical peaks were also seen when the sequence of staining was reversed. It is suggested that in the conventional Feulgen-type reactions only some of the DNA-aldehyde molecules are stained; the remaining molecules can be stained by sequential application of another Schiff or Schiff-type reagent such as acriflavine-SO2. The possible mechanism of staining in these cases has been discussed.


Studies on the Indian sand lobster Thenus orientalis (Lund): mucopolysaccharides of the tegumental glands. The tegumental glands are imbedded in the connective tissue below the epithelium of oesophagus. Each gland is made up of cells which are conical in shape with their narrow ends directed towards the lumen of the gland. In the centre of each gland there is a cavity which communicates with intracellular duct. Similar glands have been found in the hind-gut region also. These glands secrete both acid and neutral mucopolysaccharides and to some extent glycoproteins. The glands are charged mostly with the task of secreting weakly acidic mucopolysaccharides and neutral mucopolysaccharides which are confined to the apices and central cavity of the gland. The acidic nature is partly due to sialic acid and partly due to hyaluronic acid. These weak acids do not seem to play any role in digestion but lubricate the lumen of the oesophagus for easy passage of food and keep the lining of the oesophagus slimy. In the hind-gut they help in binding the faecal matter into pellets.


Cytogenetic examination of the NOR activity in a proband with 13/13 translocation and in her relatives. NOR activity in a proband with 13/13 translocation and in her relatives was examined by NOR silver impregnation and by determination of the association frequencies. In the proband, besides the fused chromosomes 13, also a chromosome 14 and a 15 showed no NOR staining. Therefore the possibility could be ruled out that the loss of NORs was compensated by the activation of inactive NORs. However, in the proband, one chromosome 22 seemed to be more intensively stained by silver nitrate than in her parents. As in the proband, the association frequency remained constant because of an increased association tendency of chromosomes 22. The possibility is discussed that the loss of NORs was compensated by a higher NOR activity of one chromosome 22.


Prenatal diagnosis of Meckel syndrome.

Prenatal diagnosis of Meckel syndrome. The three main features of Meckel syndrome are encephalocele, polycystic kidneys, and polydactyly. Prenatal diagnosis of a fetus with Meckel syndrome was made in the 16th week of gestation by means of amniotic fluid alpha1 fetoprotein estimation. The indication for amniocentesis was a previous child with an occipital meningocele and polycystic kidneys. Interpretation of the alpha1-fetoprotein value (240 microgram/ml) was difficult due to fetal blood contamination. Prenatal diagnosis is indicated in any pregnancy following the birth of a child with only two major symptoms of Meckel syndrome.


Reactions of rabbit antisera to human beta2-microglobulin (beta2-M) with platelets. Reactions of rabbit antisera to human beta2-M against human platelets and man-mouse hybrid cells were studied by MA tests and cytolysis tests. Evidence is presented that the beta2-M act as species-specific cell surface antigens, some of which are most likely shared by cell surface proteins determined by genes on chromosomes other than the chromosome 15.


Human triclonal anti-IgG gammopathy. II. Determination of the antigenic specificity patterns of the IgG, IgA and IgM autoantibodies for the subclasses of IgG. The specificity and reactivity patterns of monoclonal IgG, IgA and IgM anti-IgG autoantibodies isolated from the serum of one patient (Gil) have been determined for IgGs of the four gamma chain subclasses. The haemagglutination produced by the interaction of the Gil anti-IgGs and anti-Rh IgG coated erythrocytes was inhibited by a panel of intact IgGs, their polypeptide chains, and enzymatic fragments which included purified heavy chain constant region domains. Intact IgG1, IgG2, and IgG4 produced the same patterns of reactivity with the Gil anti-IgGs. When partially reduced and alkylated IgG1 heavy chains and its tryptic digests were tested, these were much more reactive than Fc fragments isolated from IgG of the four subclasses which were weaker inhibitors, and gamma chain constant region domains which were totally non-reactive. In all instances and by use of two anti-Rh antisera, the specificity patterns obtained for the Gil anti-IgGs were identical. The data combined with previous knowledge of the identity of the Gil light chains suggests that the antibody combining sites of these molecules are very similar if not identical.


Human triclonal anti-IgG gammopathy. III. Determination of the clonal persistence of the IgM autoantibody for a four and one-half year period. The persistence in the structure of IgM anti-IgGs obtained from the serum of a patient (Gil) with Felty's syndrome was studied. Co-isoelectric focusing experiments showed that intact IgM from early (1970) and late (1973) sera, and their heavy and light chains had identical isoelectric points (pI). Light chains from 1970, 1973, and 1974 IgM-Gil had identical amino terminal amino acid sequences and belonged to the v Kappa II light chain subgroup. Specificity studies of IgM-Gil isolated from serum obtained in 1970, 1972 and 1974 showed identical patterns of reactivity with IgG and IgG subunits of all gamma chain subclasses. Finally, by haemagglutination-inhibition, antisera specific for idiotypic determinants of IgM-Gil-1970 were shown to be shared by IgM from 1970, 1972 and 1974 sera. The data support the notion that autoantibody-producing cell clones persist unaltered during the course of an untreated autoimmune disease, in this instance for 4 1/2 years.


Heterogeneity of Epstein-Barr virus. IV. Induction of a specific antigen by EBV from two transformed marmoset cell lines in Ramos cells. Infection of cells of the EBV-genome-negative human B-lymphoma Ramos line with viral isolates obtained from two EBV-transformed marmoset cell lines (B95/8; Nyevu) resulted in the induction of a nuclear antigen (RAM-ag) apparently different from other EBV-associated antigen complexes. This antigen is revealed by indirect immunofluorescence and shows no detectable cross-antigenicity with EBNA or any other known EBV-associated antigen. EBV-isolates from P3HR-1 cells fail to induce a similar antigen in Ramos cells although they induce EBNA. No RAM-ag was expressed, either after infection of cells of another EBV-genome-negative human B-lymphoma line BJAB with B95-8 EBV or in a series of EBV-harbouring cell lines. Thus the antigen appears to be cell-line-specific for Ramos cells. It is also induced upon infection of either B95-8 or P3HR-1 converted Ramos sublines with EBV from B95-8 cells. All human sera with RAM-ag-reactivity revealed antibodies against VCA. However, sera from patients with acute infectious mononucleosis containing high anti-VCA-antibodies did not react with RAM-ag. Seroconversion for this antigen apparently more closely coincides with the appearance of EBNA-directed antibodies.


Identification and behavior of the precipitating BLV antibodies in sera of leukotic cattle. Precipitating bovine leukemia virus antibodies isolated from bovine leukemic sera by ion-exchange chromatography, gel filtration and affinity chromatography were identified as immunoglobulins of the IgG1 subclass and IgA class. They proved to be different with regard to molecular size and electrical charge. Immunoglobulins IgG2 and IgM as well as low-molecular components ranging in the microglobulin regions (less than 4S) failed to precipitate BLV antigens. Individual bovine leukemic sera showed differing precipitating activities against p24 and gp69 antigens. Precipitating monospecific gp69 antibodies represented exclusively the IgG1 subclass. Some unidentified antigens in disrupted BLV preparations showed serological relationship to p24 and gp69, respectively.


Surface expression of murine leukemia virus structural polypeptides on host cells and the virion. The expression of murine leukemia virus structural polypeptides on the surface of cells producing exogenous Friend leukemia virus, endogenous ecotropic AKR and xenotropic BALB/c virus was investigated. Antisera to Friend virus gp71, p30, p15E, p12 and p10 were employed in a complement-dependent chromium release assay and to immunoprecipitate lactoperoxidase iodinated surface polypeptides prior to analysis in polyacrylamide gel electrophoresis. With the latter technique gag-gene encoded proteins and their precursors were not discovered on the viral and cellular surface membranes. Only env-gene encoded polypeptides gp85, gp71, and p15E were detectable. p15E is embedded into the lipid membrane. gp85 is formed by disulfide linkage of p15E to surface-exposed gp71. The ratio of gp71 to gp85 is variable and apparently determined by the host cell. Antibodies of strong cytotoxicity are those against type- and group-specific epitopes of gp71 as well as type-specific epitopes of p12.


Specific cytotoxic lymphocytes against syngeneic tumors are generated in culture in the presence of syngeneic, but not xenogeneic, serum. Experiments were performed to test the effect of xenogeneic (fetal calf) serum (FCS) , as compared to syngeneic mouse serum (SMS) on the generation in culture of specific cytotoxic lymphocytes (CL) against syngeneic tumors. Sensitization in FCS against 3LL tumor cells resulted in CL cross-reacting with B-16 tumor cells and vice versa. Anti-syngeneic fibroblast CL also cross-reacted with 3LL. Such cross-reactivities were shown to be derived from CL directed against FCS determinants. In contrast, sensitization in the presence of SMS resulted in CL directed against tumor-specific antigens. Anti-3LL generated in SMS lysed 3LL targets but not B-16, and anti-B-16 lysed B-16 but not 3LL. The two types of CL had two distinct reactivities in vivo. Anti-3LL CL generated in FCS enhanced tumor growth in vivo, whereas anti-3LL CL generated in SMS had an inhibiting effect on the growth of tumor cells. These results indicate that the application of syngeneic serum during in vitro sensitization against syngeneic tumors may open up new possibilities for the analysis of tumor-specific antigens and for eliciting specific immune reactions against such antigens.


Expression of oncofetal antigens on murine sarcomas characterized for expression of endogenous MuLV. A rabbit antiserum raised by repeated immunization with BALB/c fetuses obtained at 10-14 days of gestation was used to search for oncofetal antigens (OFA) in murine sarcomas which had previously been characterized for the expression of endogenous murine leukemia virus (MuLV). Iodinated protein A from staphylococcus aureus (IPA) was used to quantitate binding of the antiserum to cultured tumor or fetal cells or to saline extracts of tumors and fetuses. Use of the "antigen" extracts facilitated the assay: the extracts bound to plastic and served as targets for the binding assay, eliminating the need to establish tumors in culture. After absorbtion in vitro and in vivo with adult tissues the rabbit antiserum bound to day 10-14 fetal cells and extract but not to endogenous MuLV (BALB virus 1). The antiserum bound equally well to MuLV-negative and MuLV-positive sublines of MCA-induced sarcomas 1420 and 1414 but not to Moloney sarcoma cells and MCA-induced sarcoma 1386. Thus, the absorbed antiserum detects a class of common cross-reacting antigens which are serologically distinct from MuLV-associated antigens.


Delayed hypersensitivity reactions of cancer patients to antigens on lymphoid cell lines. Four-hundred and fifty nine cancer patients were skin tested with extracts from five lymphoid cell lines. More than 50% of patients with lymphoma had positive skin tests with the extracts prepared from the cell line derived from Burkitt's lymphoma (BL) and more than 50% of nasopharyngeal carcinoma (NPC) patients reacted to the NPC-derived cell line extracts. Although the significant association between patient diagnosis and orgin of cell lines suggested that tumor-associated antigens were responsible for the pattern of delayed hypersensitivity, problems in standardization of antigen potency and non-specificity need to be resolved before this in vivo assay achieves its full potential.


Immunological characterization of the low-molecular-weight DNA binding protein of mouse mammary tumor virus. p14, a low-molecular-weight MMTV protein previously identified as having DNA-binding properties and encoded by the gag region of the MMTV genome, was purified by affinity chromatography on DNA-sepharose. Immunological characterization of the purified protein showed that MMTV p14 shares no cross-reactivity with gp52, gp36 and p10, antigens associated with the MMTV envelope, nor with p27 antigen found in the virion core. Purified MMTV p14 did show cross-reactivity with purified intracytoplasmic A particles, supporting the concept that A particles are morphological precursors to MMTV cores. In addition, shared antigenic determinants between intracytoplasmic A particles and MMTV p27, p20 and p10 were demonstrated. MMTV p14 did not cross-react with the low-molecular-weight DNA-binding proteins of MuLV or of type-C or -D viruses of higher mammals.


Combined chemotherapy for unresectable carcinoma of the lung: a randomized trial. A selected case file of 32 patients with carcinoma of the lung which had passed the stage of surgical cure was randomized in 3 arms and treated with different schedules: regimen A, adriamycin plus cyclophosphamide; regimen B, the BACON combination, slightly modified; regimen CVP, cyclophosphamide plus vincristine plus prednisone. Response rate and survival curves were analyzed. Statistical evaluation showed a significant increase in survival of B versus A, but not versus CVP, while other factors such as the initial performance status and the response rate seemed to have a marked influence on survival time. No significant correlation was observed with different histiotypes within each regimen. All three regimens compare favorably with patients who received only supportive treatment.


A contribution to immunological tumour diagnostics in urology. Fifty-four patients with malignant and other diseases of the urinary bladder and the prostate were examined for lymphocyte sensitization by means of electrophoretic mobility test (EM test). In this antigens from brain tissue (EF) and from malignomas of kidney, urinary bladder and prostate were used. Positive HEP values were obtained in 40 patients out of 42 with malignant tumours and in 5 patients out of 12 with other kidney diseases. With the corresponding tumour-associated antigens (TAA) in the EM-test, the histological tumour diagnosis could be confirmed in every case. Positive findings after using EF as well as the corresponding TAA are with great probability an indication of a malignant disease.


Axoplasmic flow during chronic experimental glaucoma. 1. Light and electron microscopic studies of the monkey optic nervehead during development of glaucomatous cupping. The anterior optic nerve and the macular region of the retina of glaucomatous eyes of five rhesus monkeys (Macaca mulatta) have been examined by light and electron microscopy. The experimental glaucoma had been induced by argon laser treatment of the anterior chamber angle. The eyes were examined 3 to 11 weeks after the onset of sustained elevation of intraocular pressure above 20 mm Hg. Severe degenerative changes were seen in eyes with higher intraocular pressure and longer duration of glaucoma. Eyes with a lesser elevation of intraocular pressure and shorter duration of glaucoma showed changes sharply localized to the axon bundles in the scleral lamina cribrosa. Accumulation of mitochondria and dense bodies occurred anterior and posterior to collagenous septae. The location of these changes is in agreement with the localization of block of axoplasmic transport identified by autoradiographic studies. It is speculated that these cytologic changes reflect blockage of axoplasmic flow in the optic nerve of eyes with glaucoma.


Localization of nervous system antigens in retina by immunohistology. Several antigens expressed in the nervous system were localized in tissue sections of developing and adult mouse retina by indirect immunofluorescence. Two antigens expressed in oligodendrocytes, basic protein of myelin, and NS-1 are not detectable. Antisera against nervous system-3, -4, and -7 antigens (NS-3, NS-4, and NS-7) give a uniformly intense reaction on all retinal cell structures. Large, external, transformation-sensitive (LETS) protein is present on blood vessels. Glial fibrillary acidic (GFA) protein is absent at birth but is found after day 4 in cells of the ganglion cell and nerve fiber layers. GFA protein--positive cells extend in parallel from inner to outer limiting membranes only at the ora serrata and around the optic nerve. No other glial elements contain GFA antigen, suggesting two distinct populations of astrocytes. Neurofilament (NF) protein--positive cells are present in the adult retina in outer plexiform and ganglion cell layers. By day 4 the ganglion cell layer and the ependymal zone have become NF-positive.


The relationship between cholera toxin receptor and the Thy-1 antigen determinants. The aim of the present study was to determine the relationship of the Thy-1 antigenic determinants to the cell surface ganglioside GM1. Exposure of either thymus or bone marrow cells to Thy-1 alloantiserum partially inhibited the binding of [125I]cholera toxin (CT) by the cells. Xenoantibodies to Thy-1 antigens failed to affect the binding of [125I]CT to mouse thymus cells. In the reverse experiments, treatment of either thymus cells or brain homogenate with CT did not alter their capacity to absorb Thy-1 allo- and xenoantibodies. It is concluded that GM1 ganglioside does not constitute a Thy-1 antigenic determinant.


Classification of rotaviruses: report from the World Health Organization/Food and Agriculture Organization Comparative Virology Program. The Reoviridae working team was established under the World Health Organization/Food and Agriculture Organization Comparative Virology Program in 1975. The generic name rotavirus has been adopted for the reovirus-like agents associated with diarrhea in man and animals, and the Nebraska calf diarrheal virus strain of bovine rotavirus has been selected as a candidate reference virus. Stocks of this virus and of gnotobiotic calf antiserum have been prepared. Antigenic differences among rotaviruses isolated from different species were recognized on the basis of virus-neutralization tests; a possible association between antigen specificity and variation in the RNA segments and structural proteins of rotaviruses was noticed.


Studies on gramicidin S synthetase. Purification of the heavy enzyme obtained from some mutants of Bacillus brevis. The heavy enzyme of gramicidin S synthetase was purified to an almost homogeneous state by a combination of ammonium sulfate fractionation, ornithine-Sepharose 4B chromatography, DEAE-cellulose chromatography, and Ultrogel AcA 22 chromatography. The enzyme was proved to be essentially homogeneous by ultracentrifugation and polyacrylamide disc gel electrophoresis. The heavy enzymes of gramicidin S synthetase from various groups of mutant strains lacking the ability to form gramicidin S were also purified to a similar extent. The sedimentation rates of the purified enzymes from a wild strain and the mutant strains (BI-3, BII-3, BI-9) were studied by analytical centrifugation and sucrose density gradient centrifugation. The enzymes from the wild strain and these mutant strains were all found to have an S20,W value of 12.2 at a protein concentration of 2.5 mg per ml. These results strongly suggest that the failure of specific amino acid activation in the heavy enzyme of these gramicidin-lacking mutants might be due to some modification at the active center of the corresponding amino acid-activating enzyme rather than to a complete absence of the amino acid-activating enzyme protein in the heavy enzyme.


Studies on gramicidin S synthetase. Purification and properties of the light enzyme obtained from some mutants of Bacillus brevis. The phenylalanine-activating and/or-racemizing enzyme, i.e., the light enzyme, of gramicidin S synthetase was purified to a homogenous state by D-phenylalanine-Sepharose 4B chromatography from a wild and some gramicidin S-lacking mutant strains of Bacillus brevis. The light enzyme obtained from a mutant strain E-1 could activate phenylalanine but not racemize it, and had no phenylalanine-dependent ATP-[14C]AMP exchange activity, whereas the same enzyme obtained from other mutants and the wild strain had all three activities. Furthermore, the light enzyme of the mutant E-1 could form only acid-labile enzyme-bound phenylalanine, while the same fraction of the wild strain carried half of the enzyme-bound phenylalanine as acid-labile adenylate and half as a acid-stable thioester. These results suggest that the thiol site of the light enzyme of mutant E-1 might be damaged.


Bone changes incident to blade-vent implants. This research was undertaken to study the dynamics of bone changes in relation to endosseous blade-vent implants with vital labeling technique. Three mandibular premolars were bilaterally extracted in six two years old dogs. Subsequently each healed edentulous space was implanted with a titanium endosteal blade. One blade in each dog had the post extending into the mouth; the other had the post cut at the level of the neck and covered by the mucoperiosteal flap. The animals received I.V. Pyrrolidin-Methyl-Tetracycline and Alizarine Red S labelings at 15 and 30 days intervals. Dogs 1, 3 and 5 were sacrificed on the 97th day; dogs 2, 4 and 6 on the 52nd day. On ground sections of the mandibular bone which included the blades under U.V. light, it was observed that the implants were surrounded by remodeled trabecular bone of cancellous nature containing the labeling lines of Tetracycline (yellow) and Alizarine (red). The bone-metal interface consisted mainly of a thick layer of non mineralized tissue of about 70 micrometer. The labeling results showed that the new bone apposition began early (at least by the 15th day) and continued for a long time, being still present at the moment of the last injection (90th day). No difference was recognized between the bone responses to the implants jutting out and those submerged. This confirms that implant failure is mainly linked to irregular traumatism and inflammatory infiltration which occurs over a more prolonged duration.


Rotational relaxation of free and protease-bound alpha2-macroglobulin. The recently described triplet probe depolarization technique has been utilized to investigated the rotational relaxation of free and protease-bound alpha2-macroglobulin. The molecular Stokes radius of the free globulin was found to be 88 A, a value which, when compared to the dry radius, indicates a high degree of hydration. The correlation time of alpha2-macroglobulin does not change after its binding with chymotrypsin, but slightly increases in the presence of plasmin. In the presence of 4 M urea, alpha2-macroglobulin dissociates into subunits and this dissociation does not lead to a release of the bound proteases.


Glucocorticoid induction of hepatic alpha2u-globulin synthesis and messenger RNA level in castrated male rats in vivo. alpha2u-Globulin is a male rat liver protein of Mr = 20,000 which is synthesized in the liver of adult male rats, secreted into the serum, and excreted in the urine. Its function is unknown. The hepatic synthesis of this protein is under complex hormonal control. We had previously shown that castration of male rats diminishes hepatic alpha2u-globulin synthesis and the level of its mRNA, and that administration of androgen to these castrated animals results in the reinduction of the synthesis of this protein and the level of its mRNA. We now report that alpha2u-globulin synthesis and the level of its mRNA can be fully reinduced in castrated males by administration of glucocorticoid alone. This induction is much more rapid than the androgenic induction and is inhibited by the glucocorticoid antagonist progesterone. Administration of glucocorticoid to intact male animals does not induce alpha2u-globulin synthesis above normal levels; however, if alpha2u-globulin synthesis has been depressed in intact male rats by pretreatment with estrogen or cyproterone acetate, the level of this protein can be reinduced by administration of glucocorticoids. The implications for the control of alpha2u-globulin gene expression are discussed.


Cholinergic stimulation of skeletal muscle alanine and glutamine formation and release. Evidence for mediation by a nicotinic cholinergic receptor and guanosine 3':5'-monophosphate. The mechanism of cholinergic stimulation of alanine and glutamine formation and release from skeletal muscle was studied using rat epitrochlaris preparations. The increased alanine and glutamine release produced by carbamylcholine (10(-6) M) was reproduced by tetramethylammonium (10(-6) M) but not by pilocarpine (10(-6) M) and was blocked by hexamethonium (10(-4) M) but not by atropine (10(-7) M). This increased alanine and glutamine release was not associated with altered muscle cAMP levels. However, carbamylcholine (10(-6) M) and tetramethylammonium (10(-6) M) did not increase levels of cGMP, 134% and 101%, respectively, and these increments in cGMP were blocked by hexamethonium but not by atropine. Carbamylcholine produced a concentration-dependent increase in cGMP levels. Methylisobutylxanthine and theophylline augmented the increased amino acid release and increased cGMP levels produced by carbamylcholine. Neither xanthine derivative alone altered alanine and glutamine release or cyclic nucleotide levels. Added cGMP increased amino acid release and the uptake of [U-14C]alanine and alpha-amino[14C]isobutyric acid. Carbamylcholine did not alter muscle phosphorylase a activity, glycogen levels, or basal adenylate cyclase activity. These data indicate that cholinergic stimulation of muscle alanine and glutamine formation and release involves a nicotinic cholinergic receptor and may be mediated by increased levels of cGMP, which in turn may result from a cholinergic stimulation of muscle guanylyl cyclase.


Metastatic tumors involving the cervical vertebrae: surgical palliation. Pain, weakness, or paralysis from involvement of the spinal cord and nerve roots secondary to invasion of the vertebrae by a malignant tumor often can be avoided or alleviated by stabilization of the spine. Twelve patients with neoplastic infiltration of the cervical vertebrae were so treated. The operation of wiring, augmentation bone-grafting, and decompression of the spinal cord was successful after conservative methods failed. Indications for operation were: (1) unremitting pain in the neck, not relieved by bracing or radiation therapy; (2) a major degree of vertebral destruction with loss, or impending loss, of support for the head; (3) collapse of a vertebral body; or (4) neural deficit from local tumor invasion. A classification of our twelve patients into three groups helped to delineate the surgical procedure needed. The value of obtaining spinal stability and a solid fusion above and below the tumor was evident in eleven patients. For almost all of their survival time, they were comfortable. Surgical treatment may not appreciably extend the lenght of a patient's survival, but it generally improves the patient's quality of life.


Specific glycoprotein changes during development of the chick neural retina. After separation of whole proteins of chick neural retina by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS), a number of glycoproteins can be detected by staining the gels with 125I-labeled wheat germ agglutinin (WGA) and other lectins. The glycoprotein patterns show both quantitative and qualitative changes between days 7 and 13 of development. Some of these glycoproteins can be separated by chromatography on columns of insolubilized lectins. These observations suggest that purification of some of these glycoproteins identified by staining with radioactive lectins would yield retinal antigens which may be specific for developmental stage and cell type.


New staining method for the detection of sister-chromatid exchanges in BrdU-labelled chromosomes. A direct Giemsa technique is described for the selective staining of Brd-U-substituted chromatin. It can advantageously be used for sister-chromatid exchange analysis and for the detailed cytological study of the mouse kinetochores. Presumptive inversions within the centromeric region of chromosomes 4 and 18 in the C57Black/6J mouse are described.


Detection of goitrin and its heptafluorobutyryl derivative by gas-liquid chromatography with electron capture, electrolytic conductivity and sulfur detectors. The separation of goitrin by two gas-liquid chromatographic columns, the response of different detectors and the use of the heptafluorobutyric (HFB) anhydride derivative of goitrin to improve sensitivity was investigated. The non-polar 3% OV-1 and the intermediate 4% SE-30/6% SP-2401 on 80-100 mesh Chromosorb W HP gave comparable results and were considered interchangeable. The sensitivities of the electron capture, sulfur 394 nm emission, chlorine and nitrogen Coulson electrolytic conductivity detectors were inadequate for goitrin per se. Chromatography and sensitivity of all detectors to goitrin were greatly improved by using by using the HFB derivative. It was possible to detect 1 to 60 ng of goitrin-HFB in standard solutions. Application of the technique to goitrin-spiked (2 ppm) milk, cleaned up by high-performance liquid chromatography, encountered no problems and was considered satisfactory for all four detectors.


Coagglutination test for identification of Pasteurella multocida associated with hemorrhagic septicemia. Two serogroups (B and E) of Pasteurella multocida associated with hemorrhagic septicemia in cattle were differentiated by a coagglutination test with antibody-coated staphylococci. The group antigen was soluble and heat stable. It was not detectable in unencapsulated cells or their heated extracts. Adsorbed onto human type O erythrocytes, it could be demonstrated by an indirect hemagglutination test with specific antiserum or by coagglutination with specific antibody-coated staphylococci. Antibody-coated staphylococci could detect the soluble group antigen in the plasma and liver extract of mice experimentally infected with P. multocida. The group antigen did not affix to mouse erythrocytes in vivo.


Specificity of exoantigens for identifying cultures of Coccidioides immitis. Forty-nine isolates of arthroconidial fungi other than Coccidioides immitis did not produce immunodiffusion lines of identity with the reference system for that fungus.


A Golgi study of goldfish optic tectum.

A Golgi study of goldfish optic tectum. A study of goldfish optic tectum was performed with rapid-Golgi, Golgi-Kopsch and a modified Golgi-Cox impregnation which proved quite suitable to impregnate cells in the middle tectal layers and to study more closely axonal properties. Fifteen cell types are distinguished, based upon the position of dendritic trees and axonal properties. Two cell types are found with dendrites in the marginal layer: type I with an axon terminating in the central gray layer and type II without an impregnated axon. Three cell types (III, IV and V) have dendrites in a single, specific tectal layer and an axon terminating within the tectum. Five cell types (VI-X) have dendrites in two horizontal planes. Two of them have myelinated axons leaving the tectum, whereas the axons of the remaining three types project to different tectal layers. While these first ten cell types have dendrites almost exclusively in the superficial half of the tectum, the remaining five types have dendrites in deeper layers too. This especially holds for the most conspicuous tectal cells (types XII and XIII), which have dendritic trees branching at three or more horizontal levels and a myelinated axon leaving the tectum, with sometimes a very peculiar course (XIII1). Also type XI has three or more dendritic trees, but its axon was not found. The numerous cells with cell bodies in the deepest tectal layer (type XIV) have dendrites and axonal terminations anywhere in the tectum, except in the most superficial and the deepest layer. However, most dendrites occur in the optic layers, whereas the axons, always originating from the dendritic shaft in the superficial tectal half, generally terminate in the middle tectal layers. Type XV cells have their soma in the deepest tectal layer as well, but their dendrites do not reach the optic layers. Per tectal lobe the following numbers are estimated: type I : 5,000-20,000 neurons; Type III : 2,500-10,000; types IV--XIII : each 500-2,000 and type XIV : 1,000,000-2,000,000. The total number of myelinated tectal efferents is estimated at 2,000-8,000. Comparison with other Golgi studies in teleosts leads to the conclusion that the tecta of these species of fish are basically similar.


Development of distinct cell types in the feline red nucleus: a Golgi-Cox and electron microscopic study. The feline RN contains neurons which fall into three size categories: giant (40--80 micrometer), medium (25--35 micrometer) and small (6--20 micrometer). These three populations of rubral neurons are distinguishable on the basis of a number of ultrastructural criteria and form classes not dissimilar from the traditional three divisions of cell types. Each of the three populations of rubral neurons can be further divided into three subgroups on the basis of a large number of configurational criteria discernible by the Golgi-Cox method. Each of these nine cell types are clearly separate, distinguishable by at least three criteria, and are found in different regions of the RN. It is shown that in the 5-day prenatal kitten, rubral neurons are already organized into the aforementioned three size categories. At this age most of the subpopulations are also distinguishable by the Golgi-Cox method. However, the giant rubral neurons (about 30 micrometer) and the medium sized cells (about 20 micrometer) are much smaller than in the adult cat. The dendrites elaborate many fine processes which emerge from multiple varicosities. The neuropil differs strikingly from that of the adult in that the vast majority of axons are small and unmyelinated. A number of changes in the RN are apparent as the kitten matures. The larger rubral cells undergo configurational changes before the smaller neurons, yet the giant cells continue to grow for a longer period of time. In the perinatal period, the extent of arborization of the dendritic trees diminishes, the number of spines decreases, and the long dendritic spines shorten. Somatic spines first appear in the giant cells at about one week after birth. In prenatal kittens, large cells frequently elaborate a tuft of fine branching processes in one region of the soma. These tufts later diminish in size and disappear by one week postnatal. Recent investigations (Pompeiano, '59; Condé, '66; King et al., '73' Sadun, '75) indicate that the RN of the cat is highly organized and very heterogenous. Afferent terminals are restricted to certain regions of certain cell types which are themselves specifically located within the RN. This specificity is apparent in perinatal kittens, despite the manifest immature appearance of the RN.


Correlation of computed tomography and serum tumor markers in metastatic retroperitoneal testicular tumor. Preliminary findings of an apparently good correlation between computed tomography and serum tumor markers--human chorionic gonadotropin and alpha fetoprotein--are presented. These findings are from an ongoing prospective randomized study evaluating the efficacy of intensive chemotherapy and cytoreductive surgery in metastatic retroperitoneal testicular germ cell tumors.


Reactivity of ragweed allergens with IgE antibodies. Analyses by leukocyte histamine release and the radioallergosorbent test and determination of cross-reactivity. Five distinct proteins with allergenic activity have been isolated from short ragweed pollen. We initially tested three of these, AgE, AgK, and Ra3, for reactivity with IgE antibodies by leukocyte histamine release and by the radioallergosorbent test (RAST). We found highly significant correlations between the reactivities of these allergens by leukocyte histamine release and by the RAST, consistent with the view that both procedures detected comparable allergenic activity. We next tested the allergenic cross-reactivity of all five ragweed allergens. AgE, AgK, Ra3, Ra4, and Ra5, by RAST inhibition. With solid-phase AgE the only nonhomologous inhibitor was AgK, which cross-reacted weakly and required a 140-fold mass excess of AgK compared to AgE. With solid-phase AgK both AgK and AgE produced significant inhibition; AgE was slightly more potent than the homologous AgK, Ra3 and Ra5 were allergenically unique, because only the homologous allergen produced 50% inhibition. Ra4 was weakly inhibited by AgE, Ra3, and Ra5 when these allergens were added in 300- to 5---fold mass excesses; this weak inhibition may represent either cross-reaction or cross-contamination. We found that RAST inhibition could be used as an assay for the individual ragweed allergens and we demonstrated the presence of all of the allergens in a whole ragweed extract. The sensitivity of the RAST inhibition assay ranged from 10 ng to 100 ng for 50% inhibition. Finally, the solid-phase ragweed allergens were used to determine the frequency of elevated IgE antibody levels in 65 patients with ragweed hay fever. Virtually all of the patients reacted with AgE (97%), while 88% reacted with AgK, 51% reacted with Ra3, 28% reacted with Ra4, and 17% reacted with Ra5. These results highlight the usefulness of the RAST as a specific and sensitive tool for immunochemical studies of allergens.


[The present state of prevention of rhesus immunisation (author's transl)]. Rhesus factor immunisation and its complications should disappear if the indications and the techniques for abolishing it are followed. The indications follow from the usual mechanism by which immunisation occurs. This is the passage of Rh positive fetal red blood cells into the maternal circulation where they are detected by Kleihauer's test. If in pregnancy uterine bleeding, of accidental trauma occur or if amniocentesis, versions, operations on the pregnant uterus are performed or intrauterine death occurs, the necessary preventive action has to be performed on a rhesus negative woman. When pregnancy comes to an end, be it because of delivery at term when every rhesus negative woman who has not been immunised has to be treated (if the infant is rhesus positive) so the same applies after spontaneous abortion, extra-uterine pregnancy and especially after therapeutic termination of pregnancy after which it is often forgotten. The technique is simple: Within 72 hours a dose of 85 micrograms of anti D globulin is enough if injected intravenously or intramuscularly. Sometimes this quantity has to be increased, however, when the Kleihauer test has shown that more than 5 ml of rhesus positive blood has passed into the maternal circulation, or if blood of the wrong group has been transfused. The dose to neutralise 1 ml of blood is 10 micrograms.


Cellular immunity in infectious mononucleosis. II. Specific reactivity to Epstein-Barr Virus antigens and correlation with clinical and hematologic parameters. Infectious mononucleosis (IM) patients, Epstein-Barr virus (EBV)-seropositive and seronegative healthy donors, and patients with other viral infections were tested for lymphocyte blastogenesis (LB) with phytohemagglutinin and six EBV (virus concentrate, culture supernatant, and soluble [S] antigen) or control antigens. Fluorescent antibodies to EBV viral capsid antigen of IgG, IgM, IgA specificities, to nuclear antigen (EBNA), and heterophile antibodies were also assayed. These were correlated with clinical parameters (fever, pharyngitis, adenopathy, hepatitis, splenomegaly, atypical lymphocytes, and total mononuclear cell counts). EBV viral and S antigen-induced LB was significantly greater in seropositive donors. IM patients had antigenspecific LB below that of seropositive donors initially and low responses for the acute phase of illness when clinical symptoms were present and antibody titers were maximal. Specific LB rose to a peak at 3.5 to 9 weeks when the patients had recovered, most laboratory findings had returned to normal, and antibodies had declined. At peak, specific LB in IM patients exceeded that of seropositive donors, but later declined. These results demonstrate specific cell-mediated immunity (CMI) to EBV, and indicate that this develops slowly in IM and contrasts with the evolution of the clinical events and humoral immunity. This correlation supports the hypothesis that CMI is the mechanism of terminating lymphoproliferation in IM.


Genetic control of tolerance induction to human gamma-globulin in mice. Susceptibility to tolerance induction with monomeric human gamma-globulin (HGG) was tested in different inbred strains of mice. The results indicated a differential tolerance susceptibility among the strains and that the basis for the variation is genetic in nature. By using a protocol that permits genetic analysis, F1, F2, and backcross generations of the parental strains SJL/J and C3H/Bi were examined. A multigenic control model by H-2-linked and non-H-2-linked genes showing Mendelian autosomal inheritance is proposed.


Radioimmunochemical characterization of hemoglobins Lepore and Kenya: unique antigenic determinants located on hybrid hemoglobins. Antisera were produced in rabbits to the three known types of Lepore hemoglobins, which contain hybrid delta-beta non-alpha-chains, and to hemoglobin Kenya, which has a hybrid gamma-beta non-alpha-chain. By using a sensitive radioimmunoassay technique, the absorbed antisera were shown to contain an antibody population that was specific for the hybrid hemoglobin and did not cross-react with normal hemoglobins. However, with the absorbed Lepore-specific antisera, the three known types of Lepore hemoglobins were antigenically indistinguishable from each other, suggesting that antibodies are not produced to the primary structural differences which define the three non-alpha-chains of the Lepore hemoglobins. These studies demonstrate that the non-alpha-subunits of hemoglobins Lepore and Kenya possess unique antigenic determinant sites, evidently resulting from an altered polypeptide conformation.


Cellular basis for a hapten-specific state of tolerance induced in vitro. A hapten-specific unresponsive state was induced in vitro by the incubation of normal murine spleen cells with highly conjugated dinitrophenylated bovine gamma-globulin (DNP-BGG) or a dinitrophenylated copolymer of D-glutamic acid and D-lysine (DNP-D-GL) for 24 hr. After this incubation period spleen cells were washed and cultured for 4 days with the thymic-independent antigen dinitrophenylated polyacrylamide beads (DNP-PAA) or the thymic-dependent antigen trinitrophenylated burro the erythrocytes (TNP-BRBC). Preincubation with either DNP-BGG or DNP-D-GL led to a specific depression of the in vitro anti-hapten plaque-forming cell response. The degree of depression was dependent upon the concentration of the tolerogen and the duration of preincubation. The response to DNP-PAA or TNP-PAA beads was depressed to a greater degree than was the response to TNP-BRBC. The cellular basis of the immunologic unresponsiveness induced by DNP-BGG was attributable to an inhibition of B cell function whereas the unresponsive state induced with DNP-D-GL was due to both a specific inhibition of B cell function and the activation of antigen-specific suppressor T cells.


Liposomes as model membrane systems for immune attack. I. Transfer of antigenic determinants to lymphocyte membranes after interactions with hapten-bearing liposomes. The interaction of human peripheral blood lymphocytes with liposomes containing DNP-aminocaproyl-phosphatidylethanolamine together with either egg yolk or dipalmitoylphosphatidylcholine has been investigated. When lymphocytes were incubated with liposomes at 37 degrees C, the aqueous compartment (86Rb+) and the lipid portion (3H-lipid) of the liposomes became cell associated to an equivalent extent. At 0 degrees C, however, the incorporation of 3H-lipid exceeded that of 86Rb+. Lymphocyte-liposome interactions were accompanied by the transfer of DNP to the surface of the lymphoid cell as measured by susceptibility to complement in the presence of anti-DNP antibody. Hapten transfer was not limited to liposome interactions with lymphocytes, but occurred also with other cells (e.g., Chang cells). Hapten transfer could also be demonstrated by susceptibility to K cell-mediated lysis. These findings suggest that liposomes may prove to be a useful vehicle for the transfer of new antigenic determinants onto cell surfaces. The implications of these findings are discussed in the context of using liposomes as targets for cell-mediated cytotoxic attack.


Release of arylsulfatase A but not B from rat mast cells by noncytolytic secretory stimuli. The net percentage of release of arylsulfatase activity from purified rat mast cells induced by rabbit anti-rat F(ab')2 was consistently only about 1/3 that of histamine. Isoelectric focusing of the released and residual arylsulfatase activities demonstrated specific release of the A type without B and a net percentage of immunologic release of arylsulfatase A equivalent to that of histamine. When the net percentage of histamine and arylsulfatase A release were nearly maximal (88 and 76%) in response to the calcium ionophore A23187, specific release of arylsulfatase B did not occur. Thus, arylsulfatase A and not B was associated with the secretory granule released from the rat mast cell by reversed anaphylaxis or the calcium ionophore. In contrast, subcellular fractionation of water-lysed mast cells yielded arylsulfatase B with the heparin- and chymase-containing granule fraction and arylsulfatase A in the aqueous fraction comprised of cell sap and granule water eluate. It may be that arylsulfatase B resides in a minor second granule, whereas arylsulfatase A is loosely associated with the predominant secretory granule of the rat mast cell.


Induction and mechanism of tolerance to bovine serum albumin in mice given total lymphoid irradiation (TLI). BALB/c mice given total lymphoid irradiations (TLI) were injected i.p. with bovine serum albumin (BSA) in saline, and challenged with DNP-BSA in complete Freund's adjuvant 6 weeks later. The latter animals made no anti-DNP antibody response as measured by a modified Farr assay, but made a normal anti-DNP response after challenge with DNP-BGG in adjuvant. Normal mice or mice given whole body irradiation were not tolerized by the i.p. injection of BSA in saline. Spleen cells from unresponsive mice (TLI + BSA in saline) suppressed the adoptive secondary anti-DNP response of sublethally irradiated syngeneic hosts given BSA-primed T cells, DNP-BSA-primed B cells, and DNP-BSA in saline. The suppressor cells were antigen specific, and were inactivated by in vitro treatment with anti-Thy 1.2 antiserum and complement. The findings suggest that soluble antigens administered to mice after TLI evoke a state of tolerance that is maintained by antigen-specific suppressor T cells. A similar mechanism may be involved in the maintenance of tolerance to allografts. These findings may have important clinical implications for patients treated with TLI for lymphoid malignancies.


Production of antisera specific for idiotype(s) of murine anti-(T,G)-A--L antibodies. Antibodies to the synthetic polypeptide (T,G)-A--L were raised in C57BL/10 and C3H.SW mice. For each strain, the anti-(T,G)-A--L antibodies from 10 animals were pooled, affinity purified on a (T,G)-A--L-Sepharose column, and used to immunize Lewis rats. The resulting rat antisera were adsorbed with insolubilized normal mouse globulin in order to remove anti-isotypic and anti-allotypic antibodies. The residual antibodies specifically inhibited the binding of (T,G)-A--L by anti-(T,G)-A--L as measured by a radioimmunoassay. The specificity of this inhibition was demonstrated as follows: 1) failure of the anti-(T,G)-A--L anti-idiotype to inhibit the binding of nuclease to anti-nuclease antibody of the same allotype; 2) failure of Lewis anti-[B10 anti-(T,G)-A--L] to inhibit C3H.SW anti-(T,G)-A--L and vice versa; 3) ability to absorb anti-C3H.SW anti-idiotypic activity on insolubilized C3H.SW anti-(T,G)-A--L but not on B10 anti-(T,G)-A--L. The same or cross-reactive idiotype(s) was present in the majority of individuals of each of these strains.


The induction of cytolytic T lymphocytes with specificity for p-azophenylarsonate coupled syngeneic cells. Conditions are described for diazonium linkage of the hapten p-azophenylarsonate (Ar) to mouse lymphocytes for the purpose of induction of syngeneic cytolytic T lymphocytes. These cytolytic effector cells do not lyse target cells that are coupled with an unrelated hapten, trinitrophenyl. Cell lysis can be blocked by the addition of anti-Ar antiserum. Only those target cells that share H-2 K or D end antigens with the stimulating cell population are susceptible to lysis. Several methods were used in an attempt to demonstrate the presence of the Ar cross-reactive idiotype on CTL induced in A/J mice.


Isolation and antigenic characterization of the product of a third polymorphic H-2 locus, H-2L. The serology, immunochemistry, and genetics of the product(s) of a third H-2 locus, H-2L (previously designated D') have been studied by using an antiserum raised in BALB/c H-2db mutant mice against tissues from the wild type strain, BALB/c. Genetic mapping studies and sequential immunoprecipitation experiments both indicate that this antiserum reacts specifically with L molecules. These results imply that an H-2L product is antigenically undetectable in BALB/c-H-2db mice and that the lesion in this mutant is confined to the H-2L and not the H-2D locus. Two new specificities, H-2.64 and H-2.65, are defined by the reactivity of anti-L serum on allogeneic cells, and the strain distribution of these specificities suggests the existence of at least three H-2L alleles. This third H-2 locls is therefore polymorphic and in view of this and other similarities to the H-2K and H-2D loci, it must be considered in any evolutionary models dealing with the origin of multiple subloci of the major histocompatibility complex.


Measurement of immunoperoxidase activity. A rapid and reproducible immunoassay for quantitation of cellular antigens. A procedure is described making it possible to obtain quantitative data with the immunohistochemical peroxidase anti--peroxidase (PAP) technique. After applying PAP on fixed fibroblast preparations, peroxidase enzymic activity is measured on solubilized immune-complexes. The results show this technique to be sensitive and reproducible. Differences in the amounts of the same antigen (alpha2-macroglobulin) are detected in differently treated fibroblast preparations. The value of the technique for the demonstration of cellular antigens is discussed.


Affinity chromatographic purification of the pregnancy zone protein. An immunospecific affinity chromatographic method for purification of human pregnancy zone protein (PZP) directly from serum is described. Highly purified goat-anti-human PZP-immunoglobulin was applied as a ligand. Recovery of PZP varied from 56--75%. The impurities constituted maximally 5--10% of the total protein in the eluate. The purification factor was approximately 100.


Cytoplasmic immunofluorescent staining: a micromethod with counterstain. A method of immunofluorescent staining of intra-cytoplasmic Ig in cytocentrifuged cells is described which requires as little as 3 microliter of conjugated antisera.


Flare and itch induced by substance P in human skin. Intradermal injection of synthetic substance P (10(-7)--10(-5) M in humans produced flare, wheal and itching. These responses were inhibited by oral pretreatment of the subjects with an antihistaminic drug (chlorcyclizine) or by local pretreatment with Compound 48/80 administered to deplete the local stores of mast-cell bound histamine. The findings indicate that the responses induced by substance P were mainly mediated by histamine released from the dermal mast cells. In contrast to previously studied histamine liberators, substance P was less potent when acting on rat mast cells in vitro than on human skin mast cells in vivo. When incubated with rat peritoneal mast cells, about 100 times higher concentrations (10(-5) M) were required to induce histamine release than in the in vivo studies on humans. It was concluded that substance P is a potent histamine liberator in human skin.


Cold urticaria: inhibition of cold-induced histamine release by doxantrazole. Thirteen patients with cold urticaria were studied to assess the effect of the systemic drug doxantrazole, which has actions resembling disodium cromoglycate, on cold evoked histamine release. The patients, all of whom developed an immediate local whealing response after cooling of the forearm, demonstrated release of histamine into venous blood draining that forearm. Following doxantrazole treatment, significant suppression of histamine release occurred. In some but not all patients this was accompanied by diminution of urtication in response to cooling. A double-blind study was carried out in 3 subjects, all of whom showed diminished cold-stimulated histamine release after doxantrazole. Two of these showed clinical improvement. Doxantrazole had no effect on erythema due to intradermal histamine, but did suppress the erythematous reaction to intradermal injection of compound 48/80. Our results suggest that doxantrazole or related anti-allergic agents might be useful in the treatment of cold urticaria.


Effect of ozone on lysosomal enzymes of alveolar macrophages engaged in phagocytosis and killing of inhaled Staphylococcus aureus. The role of lysosomal enzymes in the inactivation of inhaled bacteria by alveolar macrophages was studied in rats infected with aerosols of Staphylococcus aureus and then exposed for 5 hr to 2.5 ppm of ozone to determine whether pollutant-induced defects in phagocytic killing were associated with reduction in enzyme activity. Rates of bacterial ingestion and the activities of cellular acid phosphatase and beta-glucuronidase were measured simultaneously in in situ perfused right lungs by sequential staining of frozen sections for enzyme and bacteria. Quantitative measurements of enzyme activity within macrophages without ingested bacteria were made with a computer-controlled cytospectrophotometry system. Exposure to ozone resulted in diminished rates of bacterial clearance and ingestion, large increases in numbers of intra- and extracellular staphylococcal microcolonies, and an absence of enzyme activity for macrophages containing bacterial microcolonies. Enzyme activity was unimpaired in macrophages without ingested bacteria. These results, in which absence of enzyme activity occurred only in macrophages subjected to the dual insults of ozone exposure and ingested bacteria, prove a relationship between impairment in bactericidal capacity and cellular activities of lysosomal enzymes.


Antigen-specific serotyping of Neisseria gonorrhoeae. I. Use of an enzyme-linked immunosorbent assay to quantitate pilus antigens on gonococci. Purified pili from Neisseria gonorrhoeae were used in an enzyme-linked immunosorbent assay (ELISA) to quantitate human or rabbit antibodies to pili; amounts of pilus antigen on different gonococci were quantitated, and yields of pili during purification were determined in ELISA by the degree of inhibition of optical density. The amount of pilus antigen expressed on the surface of colony type 1 or 2 gonococci of three different strains varied from 450 to 9,000 ng/600 microliter of a 200-Klett unit suspension. The quantity of pilus antigen was correlated directly with the extent of piliation as determined by electron microscopy. No pilus antigen was found by ELISA in colony type 4 organisms (devoid of pili) of three different strains. No more than 10% shared antigenicity was observed for antigenically different pili. Present purification procedures for gonococcal pili provide a yield of approximately 15%. ELISA may allow better evaluation and quantitation of the potential roles of antibody to pili in the killing or opsonization of gonococci or in the inhibition of gonococcal attachment to human cells.


Clinical trial of intra-arterial polychemotherapy in the treatment of carcinoma of the oral cavity. Following studies of problems regarding the phenomenon of synchronization and cell recruitment, the authors present the results obtained using antimitotic polychemotherapy (VCR, MTX, BLM, ADM or MTC) administered to 47 patients with squamous cell carcinoma of the oral cavity. This treatment resulted in a regression in 29/47 patients (61.7 %). All patients were subjected to X-ray therapy, while only a part of the responders were subjected to surgery. Survival after 18 months from the beginning of the treatment has been 24/47 patients (51.0%). Since the majority of patients were beyond the limits of surgery and/or radiotherapy, this can be considered as a positive result, even though the risk of death due to polychemotherapy has been rather high.