Tumours of bones and joints.

Tumours of bones and joints. Tumours of bones and joints are not infrequent in dogs but are rare in other domestic animals. In the dog, most bone tumours are malignant; osteosarcomas are by far the most frequently encountered tumours, especially in giant breeds and boxers. The following main categories of bone tumour are described: bone-forming, cartilage-forming, giant cell, marrow, vascular, miscellaneous, metastatic, unclassified, and tumour-like lesions. The tumours of joints and related structures are classified as synovial sarcomas, fibroxanthomas, and malignant giant cell tumour of soft tissues.


Mercury in hair as an indicator of total body burden. Hair is known to concentrate mercury, and in general the concentration of mercury in hair is proportional to and many times higher than its concentration in the blood. The variation of the mercury concentration in human head hair was used to follow the history of poisoning in people who ingested grain treated with methylmercury. Instrumental neutron activation analysis was used for mercury determination. The initial rising slope of mercury concentration along the hair was proportional to the daily intake of methylmercury per kg of body weight. The ratio of the concentration of methylmercury in human head hair to the average body concentration was found to be about 137. By using this ratio and measuring mercury concentration in hair, the total body burden can be calculated. In the patients studied, the peak body burden ranged from 0.8 to 4.4 mg/kg in cases showing mild symptoms, from 1.5 to 6 mg/kg in cases with moderate symptoms, and from 3 to 12 mg/kg in cases with severe symptoms. The curve of the variation in mercury concentrations along the hair was also used to calculate the biological half-life of methylmercury in man. Forty-eight cases were studied and it was found that the frequency curve (population distribution curve) was grouped into two distinct regions. In about 90% of the population the biological half-life of methylmercury was 35-100 days, and 10% showed high values of 110-120 days.


Environmental contamination by mercury in Iraq. Following the outbreak of organomercury poisoning in Iraq, an investigation was carried out during the spring and summer of 1972 to evaluate environmental contamination by organomercury.Analyses were performed on fish of several species (not typical predators) and on a few specimens of aquatic birds (not fish-eating). Most fish samples contained concentrations in muscle ranging from 0.01 to 0.15 mg/kg. These concentrations are within the range found in tropical rivers. Higher figures, 0.3-0.5 mg/kg, were found downstream from a caustic soda plant south of Baghdad. A few cases of very high mercury concentrations (25-30 mg/kg) were reported from an area where fish kills had occurred. Aquatic birds (ducks and waders) contained low concentrations of mercury (900-2750 ng/g). Tail feathers of seed-eating birds were found to contain 13 500-21 000 ng/g of mercury, which is about 10 times higher than values reported from Ethiopia and within the range found in Sweden and Canada. Insect-eating birds contained 1850-5200 ng/g, which is thought to be slightly elevated. Extremely high concentrations of mercury were found in muscle tissue of dead seed-eating birds (15 000-40 000 ng/g), while feathers contained similar concentrations (9000-52 000 ng/g). These extremely high concentrations were found only in the vicinity of storehouses where treated seed was kept. No birds of prey could be caught and analysed.


Experimental studies on organomercury poisoning in chickens in Iraq. For 164 days mature chickens received a daily diet containing 40 g of wheat treated with organomercurial fungicide and 80 g of untreated diet. A control group received 120 g of normal diet daily. The experimental group was then split - half receiving the same diet for a further 155 days, by which time all had been killed or had died, the other half being fed the same diet as the controls. No cases of mercurialism were seen in the experimental group, although 2 died after 143 and 319 days respectively. Egg production fluctuated as between the groups. Mercury levels in egg albumin showed high peaks after 98 and 108 days. After the feeding of contaminated wheat had been discontinued the albumin mercury level did not fall consistently below 0.5 mg/kg for 127 days. Levels in egg yolk always remained below the level in wheat and dropped to zero 49 days after feeding of the dressed wheat had been discontinued. Mercury levels found in tissue were similar to those described by other authors. Levels in the edible parts of the single cock examined exceeded those in the hens. Implications of these findings for public health decisions on the use of chicken and eggs as food when the birds have had access to contaminated grain are discussed.


Indirect sources of mercury poisoning in the Iraqi epidemic. Secondary sources of intoxication with methylmercury poisoning other than home-made bread were explored. Mercury levels in various food items were below 0.05 mg/kg.Over 30 000 specimens of meat and internal organs of sheep were analysed because of the strong suspicion that the animals had been fed treated barley. It was found that 6-12% of the specimens analysed contained more than 0.5 mg/kg of mercury. Consumption of meat did not appear to constitute a danger of intoxication during the outbreak.


Misuse of organomercury fungicides in Brazil. A methoxyethyl mercuric compound, licensed in Brazil as a fungicide for seed dressing, was widely used for spraying tomato and other vegetable crops in 1966 and 1967.Mercury residues ranging from 0.05 to 0.30 mg/kg were detected in 14% of the tomatoes, 13% of other vegetable samples, and 57% of the tomato paste batches.A wide campaign through newspapers, radio, and television was quickly established to educate farmers and the population in general on the hazards of ingestion of mercury-contaminated food.All samples of tomatoes and other vegetables positive for mercury (residues above 0.05 mg/kg) were destroyed. Inspection posts on the main roads and highways prevented the delivery of contaminated food to cities and food processing plants.The sale of organomercury fungicides is now controlled in Brazil; however, treated seeds which are not planted may be diverted to human or animal consumption. There is no official record of accidents, but occasional cases of poisoning are known.The mercury residue level in Brazilian fish is low, indicating that environmental pollution by mercury is not a big problem in the country. However, some fresh-water and estuarine fish may contain mercury residues ranging from 0.01 to 0.66 mg/kg.


An outbreak of organomercury poisoning among Iraqi farmers. An outbreak of organomercury poisoning due to the consumption of treated grain by farmers and their families occurred in Iraq in 1971-72. A total of 6530 cases were admitted to hospital and of these 459 died. However, there were many more with minor symptoms of poisoning who consulted outpatient departments. This outbreak constituted the largest poisoning epidemic ever recorded. No age was exempt and no pronounced sex difference was apparent. The latent period of up to 60 days between dosage and the onset of symptoms was probably the major factor contributing to the size of the epidemic. Measures taken to limit the outbreak are outlined.


Epidemiology of organomercury poisoning in Iraq. I. Incidence in a defined area and relationship to the eating of contaminated bread. A survey was carried out in a defined area in rural Iraq where there had been many cases of organomercury poisoning following the consumption of bread contaminated by mercury, in order to determine the true incidence of the disorder. The results were compared with those obtained from a similar rural area from which few cases had been reported. A questionnaire was used to determine the amount of contaminated bread eaten and the frequency of symptoms; a simple clinical examination was performed and blood and hair samples were collected for estimation of total mercury concentration. Of 700 people over the age of 5 years in the high-exposure area, 66% admitted to having eaten contaminated bread, while none of the 864 persons in the low exposure area had done so. The mean period during which contaminated bread was eaten was 32 days, but some people had eaten it for as long as 3 months. A mean of 121 loaves was eaten, the maximum being 480 loaves. For the mean number of loaves the intake of methylmercury was likely to have been between 80 mg and 250 mg, but the people who had consumed the largest amount of contaminated bread may have ingested up to 1 000 mg of methylmercury over a 3-month period. Of those with signs of alkylmercury poisoning at the time of the survey, 80% had eaten more than 100 loaves, and 53 (71%) out of 75 persons who had eaten more than 200 loaves showed some evidence of poisoning.The incidence rate for poisoning was estimated at 271 per 1 000; this figure includes a mortality rate of 59 per 1 000, 32 per 1 000 cases with severe disability, 41 per 1 000 cases with mild or moderate disability and 138 per 1 000 cases with only subjective evidence of poisoning at the time of the study.


Epidemiology of organomercury poisoning in Iraq. II. Relationship of mercury levels in blood and hair to exposure and to clinical findings. In the survey described by Al-Mufti et al. (see page 23) blood and hair samples were analysed for total mercury by modified atomic absorption spectrophotometry. The hair samples were divided into 2.5-cm segments and analysed consecutively. The mean blood levels were 34 ng/ml and 7 ng/ml, respectively in those who had and those who had not eaten contaminated bread.Corresponding mean maximum hair mercury values were 136 mug/g and 5 mug/g, respectively. Hair mercury values provided a better discrimination between different categories of exposure than blood mercury values at the time the survey was performed, some months after the end of the outbreak. Those persons who had not eaten contaminated bread but who lived in the area of high exposure had hair mercury values between the values of those who had eaten and those who had not eaten contaminated bread and who lived in the area of low exposure. Sequential estimation of mercury in 2.5-cm segments of hair in women gave information on the period of accumulation of mercury more than 1 year before the time of collection of the samples. It was possible to show an approximate relationship between the maximum hair mercury value and the amount of contaminated bread eaten. The match between the blood mercury level and the severity of poisoning was poor, owing to the length of time that had elapsed between the onset of poisoning and the sampling. With hair mercury, while the group results showed a good relation to the severity of poisoning, in individual cases the match was less good, especially in those persons where an insufficient length of hair was available for analysis. Biological variation in sensitivity to methylmercury was also likely to have been an important factor.


Epidemiology of organomercury poisoning in Iraq. III. Clinical features and their changes with time. Three categories of mercury poisoning were defined in the survey described by Al-Mufti et al. (see page 23) and the age-specific incidence rates for these are given. Persons with physical signs consistent with a diagnosis of organomercury poisoning were allocated to categories of severe disability or mild/moderate disability. However, the largest category consisted of persons who had symptoms but no readily elicitable physical signs at the time of the survey. These symptoms followed a consistent pattern with paraesthesia involving the lips and/or circumoral region or trunk and difficulty with walking, described as weakness or unsteadiness of the legs, and in some cases repeated falls, forming the most commonly occurring symptom complex. Mean maximum hair mercury levels differentiated this group very clearly from the group with no symptoms of mercury poisoning. Very few people in the area of low exposure complained of such symptoms; where they did occur they were less well related to the time of the outbreak and showed little tendency to improve. Most people reported improvement in their symptoms by the time of the survey, with more improvement in some symptoms than in others. However, it is not known whether those people with symptoms only at the time of the survey had had at an earlier stage mild signs which had cleared. It was thought unlikely that further substantial improvement would occur in those persons with disability at the time of the survey.


Chemical type of mercury in patients in the outbreak of organomercury poisoning in Iraq. Gas chromatography was employed to identify the mercurial compound involved in the Iraq outbreak of organomercury poisoning. Only methylmercury was detected in human blood, hair, and tissues. The quantity of methylmercury present correlates well with the quantity of organic mercury determined by selective atomic absorption.


Intoxication due to alkylmercury-treated seed--1971-72 outbreak in Iraq: clinical aspects. Sixty-six hospitalized patients suffering from chronic methylmercury poisoning were examined in Baghdad during 1972. The poisoning was attributed to consumption of home-made bread prepared from seed wheat treated with mercurial fungicide. The age incidence ranged between 4 and 70 years.Of the various clinical features encountered, neurological symptoms and signs were predominant and included muscular weakness, numbness, unsteady gait, paraesthesia, dysarthria, mental disturbances and, in severe cases, blindness, partial deafness, stupor, coma, and death. Involvement of the cardiovascular, urinary, gastrointestinal and haemopoietic systems, which was commonly encountered in ethylmercury poisoning in the 1960 outbreak in Iraq, was unusual.The severity of symptoms and signs was, broadly speaking, dose-dependent; high exposure led to severe clinical manifestations, but variations existed. Criteria, based on the clinical manifestations, were set for grading the severity of cases. The series included 2 asymptomatic cases, 20 mild, 20 moderate, 14 severe, and 10 very severe. In the latter group 5 patients died from failure of the central nervous system.The severely poisoned patients died irrespective of the medical treatment received. After 2 years of observation, most patients graded as mild or moderate cases lost their symptoms completely. Severe cases improved slowly, although ataxia, diminution of visual field and acuity and paraesthesia were still present. Thus, the previously accepted view that neurological signs were irreversible has been disproved.


Ocular manifestations of mercury poisoning. One hundred patients suffering from organomercury poisoning who were hospitalized in the Medical City, University of Baghdad, were examined ophthalmologically in the period between March and December 1972, and were reviewed again about 10 months later.The examination included clinical signs and symptoms, slit lamp examination of the anterior segment of the eyes, examination for visual field changes and, in the autopsied cases, analysis of the optic nerve and the occipital cortex. In addition, aqueous humour was aspirated in 10 patients and in 10 controls for the estimation of the mercury level.


Clinical observations on treatment of alkylmercury poisoning in hospital patients. Twenty-six patients suffering from methylmercury poisoning were treated by different therapeutic regimes. Seven received penicillamine or N-acetyl-d-L-penicillamine, 10 were treated with dimercaprol sulfonate and 9, who could not be treated, were given a placebo and were used as controls. Penicillamine, N-acetyl-d-L-penicillamine, and dimercaprol sulfonate reduced the blood level of mercury and increased its urinary excretion. No appreciable clinical improvement was noticed among the severe or very severe cases, while mild or moderate cases improved clinically irrespective of the treatment given.


Tissue levels of mercury in autopsy specimens of liver and kidney. Fifty-one autopsy specimens of liver were analysed for total mercury. Thirteen specimens contained less than 10 mg/kg of mercury, with a minimum value of 1.4 mg/kg, indicating that death in suspected cases was not always due to lethal exposure to methylmercury. The methylmercury concentration in 28 livers was 10-30 mg/kg. Limited additional estimations have shown that 71% of the liver mercury was organic and that the level of mercury in the liver of a 7-month-old fetus was only 25% of that in the liver of the mother. In a patient who died in hospital with a blood mercury level of 4.1 mug/ml, the liver contained 16.5 mg/kg of mercury.Differences between these results and those found in the outbreak of methylmercury poisoning in Japan are discussed. Any extrapolation of tissue mercury levels in relation to the toxic effects of methylmercury must take account of the intensity and duration of exposure.


Levels of mercury and pathological changes in patients with organomercury poisoning. Autopsies were carried out on 4 adults who died during the outbreak of mercury poisoning in Iraq and on 4 infants who were exposed to organomercury in utero. Mercury levels in tissues and in some body fluids were determined. The high levels of mercury in the central nervous system and the marked neuronal degeneration are noted.


The determination of 5-hydroxytryptophan and its metabolites in plasma following administration to man. The estimation of 5-hydroxytryptophan and its metabolites 5-hydroxytryptamine and 5-hydroxyindoleacetic acid in plasma is described. The method is based on solvent extraction and the use of a liquid cation exchange reagent to separate 5-hydroxytryptamine and 5-hydroxytryptophan. The 5-hydroxyindoles are determined fluorimetrically following derivatisation with o-phthalaldehyde in strong acid. Data on the specificity and linearity of the method are presented. The time course of plasma 5-hydroxyindoles after oral administration of 5-hydroxytryptophan is studied in two subjects as an example of the application of the method and of its possible role in monitoring the plasma levels in therapy or in provocative tests.


Experimental study on the electrical impedance of bone and the effect of direct current on the healing of fractures. An experimental study of the electrical impedance of cortical bone and medullary cavity and on the effect of a D.C. or a periodically interrupted D.C. on the healing of fractures in rabbits, suggests that the bone behaves as a capacitance of about 20 muF in parallel with a high D.C. resistance. Also a 10 muA periodically interrupted D.C. current stimulates osteogenesis.


Osteogenic sarcoma: state of the art with high-dose methotrexate treatment. The development and application of the VCR-MTX-CF regimens for the treatment of osteogenic sarcoma have changed the biological behavior of the tumor. Recent results strongly project major advances for the future. Although the major effect of chemotherapy resides in the eradication of micrometastases, its application for treatment of the primary tumor may also be considered. However, careful experimental design and follow-up periods for several years will be required to determine the optimum approaches. For example, it is possible that the interaction between weekly VCR-MTX-CF and radiation therapy may assume increasing importance. Thus, with the effective application of VRC-MTX-CF, the management of osteogenic sarcoma has evolved into a multidisciplinary approach and future advances will be based on the collective judgement of specialists from many fields.


Ventricular size following head injury: a clinico-radiological study. A retrospective study of the records of 93 patients who had sustained a severe closed head injury was carried out to determine the relationship, if any, between the severity of the injury, the size of the cerebral ventricles and the residual disability. Ventricular size was expressed as a ventricular index. The callosal angle was also measured to see whether this was of value in the recognition of post-traumatic external obstructive hydrocephalus. A normal range of ventricular indices and callosal angles was obtained from a review of the air encephalograms of 56 patients matched for age and sex with the head injury group, and whose air studies were considered to be normal. The findings on the head injury group were subjected to detailed statistical analysis. The study showed that there was no statistically significant relationship between severity of injury, measured by duration of unconsciousness, and ventricular size expressed as a ventricular index. There was a relationship between ventricular size and residual disability as graded in this study. If age was taken into consideration, the relationship between ventricular size and disability was closer. It was not possible to predict residual disability of individual patients from ventricular size alone due to the wide range of values in any disability grade. Failure of air to pass from the basal cisterns over the cerebral convexities is a widely accepted sign of extraventricular obstruction. The shapes of the ventricles, the ventricular index and a callosal angle of 110 degrees or less provided supporting evidence of obstruction in this study. Recognition of an obstructive element in ventricular dilatation following head injury is important, since in a small carefully selected group of patients a ventricular shunting operation may favourably affect recovery.


Coils of the cochlea.

Coils of the cochlea. Abnormalities of the cochlear portion of the bony labyrinth may often be suspected but are inadequately demonstrated by conventional tomography in the antero-posterior position. Further views are necessary. The authors demonstrate this by means of a comparison between tomographic and histological sections of the inner ear of an anencephalic with a unilateral Mundini defect. Submento-vertical tomograms give a good demonstration of the cochlea.


Intermediate alpha1-antitrypsin deficiency resulting from a null gene (M-phenotype). The presence of a null gene for alpha1-antitrypsin was detected in a family study by the inheritance of intermediate antitrypsin deficiency in association with a normal (PiM) phenotypic pattern. The proband, a 42-year-old man (M-phenotype), was a cigarette smoker and had physiologic evidence of pulmonary emphysema. Three female members of the family were receiving estrogenic medication but had deficient values for serum trypsin inhibitory capacity nevertheless, indicating an unresponsive gene. The mean serum trypsin inhibitory capacity for those with an M-phenotype was significantly lower than that found with an MZ phenotype, presumably due to the total noncontribution to serum antitrypsin activity by the null gene. A quantitative measurement of antitrypsin activity or concentration is necessary in an antitrypsin screening program, since phenotyping procedures alone cannot reveal the null gene.


Occlusion of the left main coronary artery secondary to left ventricular angioma. Our experience with a patient who had an angioma of the anterior wall of the left ventricle that produced complete occlusion of the left main coronary artery is presented. Diagnosis was made before surgery from findings on cineangiographic studies. Successful surgical treatment consisted of a double aortocoronary bypass to the left anterior descending and obtuse marginal coronary arteries. The angioma was left undisturbed.


Right coronary artery dissection. A complication of cardiac catheterization and coronary angiography. Four patients are described in whom right coronary artery dissection occurred during retrograde catheterization of the left ventricle or coronary arteries. In two patients, acute myocardial infarction occurred. The possible causes, radiographic features, and clinical implications of this infrequent complication are discussed.


Aortocolic fistula, a lethal cause of lower gastrointestinal bleeding: report of a case. Aortocolic fistula occurs with spontaneous rupture of aortic and iliac aneurysms into the sigmoid colon, or due to involvement of the aneurysmal wall by acute diverticulitis. In the eight cases reviewed, this complication proved uniformly lethal, although sufficient clinical findings were present for diagnosis, and adequate time was available for a planned therapeutic approach. Lower gastrointestinal bleeding in the patient who has an aortic aneurysm and left-lower-quadrant inflammation suggests the presence of an aortocolic fistual. Angiography should be performed during a bleeding episode to confirm the diagnosis. Surgical correction consists of an axillofemoral by pass graft, excision of the aortic aneurysm, and a Hartmann procedure.


HL-A antigens in clinical medicine.

HL-A antigens in clinical medicine. Many autoimmune diseases show a significant association with one or two second segregant series histocompatibility antigens. These associations are of great scientific interest, since they support the concept of HL-A-linked immune-response genes governing specific disease susceptibility. However, with one major exception, the association of HL-A antigens with diseases is not striking enough to provide a worthwhile diagnostic test. The exception is the extraordinarily high incidence of HLA B27 in patients with seronegative spondyloarthropathy best typified by ankylosing spondylitis (AS) and Reiter's disease (RD). In patients with these rheumatic syndromes, the antigen is present in more than 90% of cases compared to an incidence of approximately 6% in normal Caucasians and 4% in black Afro-Americans. The vast majority of rheumatic diseases are readily diagnosable on the basis of a history, physical examination and careful radiographic survey. This applies to most patients with a seronegative spondyloarthropathy, especially when the disease presents as a typical and fully formed clinical syndrome characterized as AS or RD. Sometimes the initial clinical nature may be atypical and only long-term follow-up of the patient will reveal an evolution toward the typical syndrome. In these situations, the correct diagnosis is reinforced by detecting the presence of HLA B27 on the patient's cells. Examination of the patient's family often reveals a high incidence of similar clinical syndromes, nearly always associated with the presence of the antigen. Since tissue typing at the moment is an expensive and relatively unavailable laboratory technic, its widespread and indiscriminatory use as a diagnostic test cannot be encouraged. However, in the clinical settings outlined above, tissue typing provides an invaluable diagnostic test. Presently, the combination of a negative test for rheumatoid factor and a positive test for HLA B27 is one of the strongest diagnostic laboratory profiles available to the physician when faced with a patient with early or atypical rheumatic disease. Aside from the purely clinical setting, the most exciting aspect of the association between these diseases and a specific cell surface antigen lies in the hope that we have a clue to the pathogenesis of a group of common rheumatic disorders. If the cause or causes of spondyloarthropathy can one day be found, the detection of HLA B27 may provide a useful public health measure facilitating preventive medicine. Even now, the detection of susceptible subjects within a family or a population will open the way for early diagnosis and treatment.


Thyroxine (by competitive protein binding analysis) in human and cow milk and in infant formulas. In 45 mothers of full term infants the level of thyroxine (T4) in serum (45 samples) and unskimmed milk (92 samples) was estimated with the aid of competitive protein binding analysis. On the 3rd day after delivery the T4 level in serum was found to be 11.1 +/- 0.4 mug/100 ml, while that in milk was 1.3 +/- 0.3 mug/100 ml. During following days of lactation the T4 concentration in sera decreased. In contrast, however, a gradual increase of the T4 content in milk was observed. The serum and milk levels equalized during the third week post partum (7.3 +/- 1.3 and 7.6 +/- 1.4 mu6/100 ml, respectively). After the 25th day the T4 level in milk was found to be higher than in sera from the same women (paired test: P less than 0.005). Values of 12.9 +/- 1.3 in milk and 8.1 +/- 0.7 mug/100 ml in sera were found in the second month of lactation. A negative correlation between serum and milk T4 level was found during the early lactation (3-20 days), while after 20th day this correlation was positive. Very low and almost undetectable levels of T4 were found in cow's milk and infant formulas derived from it.


Specific antidiuretic effect of [Leu4, D-Arg8] vasotocin and [Mpr1, Leu4, D-Arg8] vasotocin. Comment on the idea of lipophilic properties and position 4. [Leu4, D-Arg8] vasotocin (I) and [Mpr1, Leu4, D-Arg8] vasotocin (II) possess a considerable and very specific antidiuretic effect. In I, the change of configuration in position 8 caused an increase of the magnitude of the antidiuretic effect. In I, the change of configuration in position 8 caused by two orders and a decrease of the magnitude of the pressor effect by one order. I and II are the first vasotocin analogues reported to have a relatively high and specific antidiuretic effect.


Optimization of determination of ovulation after clomiphene therapy. A total of 131 cycles of 71 sterility patients, who suffered from anovulatory cycles at least previous 6 months, were monitored daily by a rapid radioimmunoassay for LH and by gynecological examinations before we attempted to induce ovulations with Clomiphene. The width of the cervix, spinnbarkeit, transparency of the cervical mucus and karyopycnotic and acidophilic index of the vagina were recorded daily. 18 subjects exhibited hypoplasia of the uterus and 32 husbands of the 71 patients suffered from severe to medium oligospermia. Homologous inseminations were performed in 15 patients. Different patterns of LH serum levels were observed during and after Clomiphene treatment. The response of LH to Clomiphene and preovulatory LH rise was recorded in 73 cycles. No Clomiphene response with preovulatory LH peak was observed in 31 cycles. 11 patients out of 71 subjects failed to ovulate. 12 patients (30.7%) out of 39 subjects with normospermatic husbands became pregnant. 6 patients (18.7%) of 32 subjects whose husbands suffered from oligospermia became pregnant. A total pregnancy rate of 30% calculated, when the 11 patients with negative responses are excluded. The conclusion is reached that sterility patients benefit most, when both gynecological as well as rapid LH estimations by radioimmunoassay are used to time imminent ovulations in Clomiphene treated subjects.


Testosterone in human urine as determined by gas-liquid chromatography. A simple and accurate gas chromatographic method for the determination of testosterone in human urine is described. Radioactive testosterone was added to samples to correct methodological loss. A mild acid hydrolysis under protection of benzene layer gave a single estimate of testosterone glucuronate and sulphate. Paper chromatography and rechromatography on chromatoplate served for preliminary qurification. Testosterone was gas chromatographed as acetate and guantitated by the internal standard procedure and flame ionization detector. Results of control experiments, normal values for female and male subjects and estimations in patients with alterations of androgen hormone metabolism are presented.


Effect of antiandrogen (cyproterone acetate) on the action of testosterone on bone protein. The effect of castration, androgen and antiandrogen treatment on bone protein content of femoral diaphyses of male mice was investigated. Castration as well as administration of 2.5 mg of cyproterone acetate for 21 days reduced significantly the protein content of the bone. Testosterone fully compensated the bone protein depletion seen in castrates. Simultaneous administration of both cyproterone acetate and testosterone to castrates blocked the testosterone action. The data show that the mice bone matrix is a highly androgen dependent tissue.


Distribution of corticotropin releasing factor(s) activity in neural and extraneural tissues of the rat. Distribution and dose-response characteristics of corticotropin releasing factor(s) (CRF) activity in the central nervous and extraneural tissues of the rat were examined with a sensitive CRF bioassay using ACTH secretion by cultured rat adenohypophyseal cells. Dose-response curves for hypothalamus were steeper than and not parallel to those for cerebral cortex, liver, serum or human urine. The minimum effective dose was smallest for the posterior pituitary. CRF activity of the basal hypothalamus was considerably higher than that in other parts of the hypothalamus, and was unaltered by 2.5 or 10 min ether stress. Our data indicate that "specific" CRF is concentrated primarily in the basal hypothalamus and posterior pituitary and is not identical with widely distributed extrahypothalamic extraneurohypophyseal CRF.


Studies on factor VIII activation potential in hemophilia A-plasma and its significance for the comprehension of hemophilia. In a factor VIII exchange test experiment higher factor VIII activities are measurable than would be expected from the single activities of the used plasmas. The same goes for the use of plasmas from carriers of hemophilia A, but not for plasmas from patients with v.W.J.S. (von-Willebrand Jürgens syndrome). In plasmas from hemophilic adults activation obviously proceeds at a slower rate than in plasmas from hemophilic children. The experimental results lead to the hypothesis that factor VIII for its activation needs a specific activator or activator system. This process can be stopped by natural inhibitors. Contrary to existing opinion, the coagulation defect of hemophilia A is not to be sought in an inability to be activated or even in absence of factor VIII, but in a previous step, in its activator. This activator is absent or defective. In patients with v.W.J.S. disease the activator or activator system is intact, but factor VIII is missing.


Studies on the mitogenic activity of trypsin, pronase and neuraminidase on human peripheral blood lymphocytes. Human peripheral blood lymphocytes were cultured in the presence of protease (trypsin and pronase) and Vibrio choleraneuraminidase. T and B lymphocyte populations were separated and the effect of these enzymes plus phytohaemagglutin or Tuberculin was studied. The results of these experiments show that proteases moderately stimulate spontaneous deoxynucleic acid synthesis of control cells and potentiate the effect of tuberculin on sensitized cells. These enzymes act specifically on B lymphocytes. Neuraminidase also increases spontaneous deoxyribonucleic acid synthesis of control cells and augments significantly the in vitro response to PPD. There is no additive effect of neuraminidase on phytohaemagglutin stimulated cells. Neuraminidase seems to stimulate specifically T lymphocytes. Some possible mechanisms of action of these enzymes are proposed and discussed.


Generation of antibody diversity. IV. Variation within single clones of antibody-forming cells developing in vivo. Previous experimental work demonstrated that clonal variation occurs in vitro. The present experiments were designed to test for clonal variation in vivo. B cells were transferred at limiting dilution, with antigen, into irradiated recipients. Seven days later spleens were assayed for plaque-forming cell (PFC) colonies. Control experiments showed that these PFC colonies were clones, that is, they were derived from a single B cell precursor. When the clones were analyzed for heterogeneity of the PFC population, using cross-reactivity on various mixtures of red blood cells as a method of detecting differences in antibody specificity, from 23-83% of the clones contained variants. By adjusting the amounts of helper activity and antigen available to a developing clone, we have been able to influence this variation; high levels of help and/or antigen favor pure clones, while low levels of either produce mainly mixed clones.


The role of thymus-derived lymphocytes in an antibody-mediated hapten-specific helper effect. The cellular basis of the antibody-mediated, hapten-specific anti-idiotype antibody response of CBA/J mice to a hapten conjugate of the BALB/c myeloma protein LPC-1 was examined. Previous studies have shown that anti-idiotype antibody is produced if the mice are given anti-2,4-dinitrophenyl (DNP) antibody and then immunized with DNP-LPC-1; immunization with DNP-LPC-1 or LPC-1 alone does not lead to the production of this antibody. In the present experiments, mice depleted of thymus-derived lymphocytes by two different techniques failed to make anti-idiotype antibody after transfer of anti-DNP antibody and immunization with DNP-LPC-1. However, thymus-deprived mice given anti-DNP antibody and reconstituted with thymus-derived spleen cells did make anti-idiotype antibody in response to DNP-LPC-1. Thymocytes could not reconstitute such mice for this response. The possible mechanisms for this hapten-specific, antibody-mediated, T cell-dependent helper effect are discussed. It is felt that such helper effects represent an alternate pathway to conventional specific T cell-mediated cooperation. Further examples of antibody-mediated helper effects which support this hypothesis are also presented.


Spontaneous release of T cell receptors for alloantigens. II. Induction of antibodies to T cell receptors. Spleen cells from normal mice, when cultivated in vitro, released receptors or recognition structures (RS) for alloantigens into the surrounding medium. The spontaneous shedding of receptors was revealed by their ability to induce the formation of anti-recognition structure(anti-RS) antibodies upon injection into appropriate F1 hybrid recipients. Cell suspensions containing T and B lymphocytes and those containing T lymphocytes were capable of inducing anti-RS antibody formation, whereas suspensions devoid of T cells were incapable of doing so. Receptors shed from such cell suspensions during a 24-hour cultivation period gave exactly the same results. Cell-free culture supernatants, however, incited higher anti-RS antibody titers, presumably because of an accumulation of RS. The capacity of released T cell receptors to recognize alloantigens as determined in the PAR assay and their ability to induce anti-RS antibodies went roughly parallel.


Surface morphology of murine B and T lymphocytes: A comparative study by scanning electron microscopy. A variety of murine lymphocytes of known B or T derivation obtained from different lymphoid organs were prepared for scanning electron microscopy (SEM) by the critical point drying method after collecting the cells by aspiration onto silver membranes. Comparison of SEM appearances of cells prepared by this technique and serological classification according to surface antigens showed that most T cells had smooth surfaces with few microvilli, while many B lymphocytes were moderately to markedly villous. Further evidence for the above correlation was obtained by examining thymic cells and enriched B or T cell populations. Thymic cell suspensions containing less than 5% B cells showed over 80% generally smooth cells by SEM. Enriched T cell populations, obtained by mass cytolysis of lymph node preparations with anti-Ia or anti-Ig sera or by purification through nylon fiber columns, contained over 85% T cells, and more than 75% of them were of the smooth cell type. A similar correlation was noted for enriched B cell populations obtained by cytolysis of lymph node cells with anti-Thy-1 serum, and by lysis of EAC-rosettes. Over 90% of these cells were identified as B cells by immunologic methods and approximately 75% had moderate to markedly villous surfaces. The 15% difference can be accounted for by the existence of a subpopulation of smooth B cells. Direct observation of EAC-rosettes confirmed that most B cells had moderate to large numbers of surface microvilli and that less than 10% were smooth. It is possible that some of the smooth cells seen in enriched B cell populations may represent precursors or B lymphocytes at different stages of differentiation. These results indicate that murine T and B lymphocytes, like their human counterparts, can be recognized in many cases under the SEM on the basis of their surface morphology. Smoother B and more villous T cells are difficult to classify by SEM without parallel immunologic identification.


The effect of specific cell inactivation on the cellular immune response to ferredoxin peptides. Previous studies using synthetic immunogenic molecules containing two haptenic peptides (ther NH2-terminal heptapeptide and the COOH-terminal pentapeptide of oxidized ferredoxin (O-Fd) from C. pasteurianum) have shown that both peptides individually are capable of initiating lymphocyte transformation and inhibiting migration in populations of lymphocytes from O-Fd-sensitized guinea pigs. While migration inhibition could readily be stimulated by single haptenic peptides, lymphocyte treasformation appeared to be more easily induced by molecules containing two or more haptenic peptides (these could be identical or different) (Kelly, B., Levy, J.G. and Hull, D., Eur. J. Immunol., 1973. 3:574). If lymphocyte transformation is a T cell-mediated phenomenon, these observations indicate the possibility of T cell-T cell interaction. The two haptenic peptides (designated "N" and "C") were synthesized and conjugated to succinylated bovine serum albumin (S-BSA), forming the conjugates N-S-BSA and C-S-BSA, respectively. These conjugates were labeled to high specific activity with 125iodine and were used in an "antigen suicide" procedure to treat guinea pig lymph node cell preparations previously sensitized to O-Fd and keyhole limpet hemocyanin (KLH). Cell populations exposed to either 125I-labeled C-S-BCA demonstrated decreased lymphocyte transformation in the presence of O-Fd but not in the presence of KLH. These results indicate specific cell inactivation by the radioactive peptide conjugates of those cells responsible for initiating cell transformation. Experiments performed by mixing 125I-labeled N-S-BSA-treated cells with 125I-labeled C-S-BSA-treated cells were successful in partially restoring the response to O-Fd and suggest possible synergy between N and C determinant binding cells in the cellular immune response to O-Fd. Evidence from B cell depletion studies suggests that this is a T cell-T cell interaction.


Receptors for IgG molecules on human lymphocytes forming spontaneous rosettes with sheep red cells. A mixed rosette technique with sheep red blood cells (SRBC) and ox erythrocytes heavily coated with rabbit antibody (EA) was employed to simultaneously identify human peripheral blood T lymphocytes and IgG receptor-bearing cells. The findings of a noticeable proportion of "mixed rosettes" in peripheral blood lymphocytes freshly drawn from normal individuals and of an even higher number of these mixed rosettes in cell suspension kept in culture media supplemented with fetal calf serum provide evidence for the capacity of human T cells to express membrane receptors for antigen-antibody complexes.


"Natural" killer cells in the mouse. II. Cytotoxic cells with specificity for mouse Moloney leukemia cells. Characteristics of the killer cell. Normal mice contain cytolytic cells with specificity for in vitro grown mouse Moloney leukemia cells. Such killer cells are most frequent in the spleens; lymph node and bone marrow contain less and thymus virtually no killer activity. Peak activity is found around one to three months of age. Spleen cells from genetically athymic mice are as active killer cells as those from normal mice of the same strain. Treatment with anti-theta serum plus complement followed by removal of adherent and surface Ig positive cells by filtration through anti-Ig columns will leave between 1-5% of the original spleen cell population from a normal mouse. These cells have the morphology of small lymphocytes and perhaps contain all of the total original killer activity of the spleen against the Moloney leukemia cells. Such killer enriched cells are devoid of T and B lymphocytes and largely fail to function in antibody induced, cell-mediated lysis against antibody-coated chicken erythrocytes. It is concluded that the spontaneous selective cytotoxic activity of normal mouse spleen cells against Moloney leukemia cells is exerted by small lymphocytes of yet undefined nature.


Cell-medicated cytotoxicity against ectromelia virus-infected target cells. III. Role of the H-2 gene complex. The role of the H-2 gene complex in expression of cytotoxicity exerted by specific ectromelia-immune thymus-derived (T) cells against ectromelia-infected target cells was examined. A repertoire of inbred mouse strains (some congenic) including the H-2 haplotypes k, d, b, s, q, the recombinant H-2a(k/d) and F1 hybrids (k/b and d/b) were immunized with virus and their spleen cells tested 6 days later, at the peak of the primary response, against H-2k,H-2d and H-2b target cells. Significant specific cytotoxicity occurred only when the immune cell donors and the target cells shared all or part of the same H-2 gene complex. For example, H-2a (k/d) immune cells killed both H-2k and H-2d target cells. There was no detectable effect of the non-H-2 genetic background, H-2 public specificities, or the M-locus. Target cells infected with ectromelia virus exhibited quantitative or qualitative changes (or both) in expression of normal H-2 antigens as indicated by reduced susceptibility to killing by T cells activated against H-2 antigens in mixed lymphocyte culture. These data are consistent with the hypothesis that T cells in this system are responding to virus-induced, specific changes in antigens on infected cells which are controlled by genes in the H-2 complex; these genes seem likely to be those coding for H-2 private specificities, or genes closely linked to them.


Five types of lymphocytes (Ig-theta-, Ig-theta+weak, Ig-theta+strong, Ig+theta- and Ig+theta+) characterized by double immunofluorescence and electrophoretic mobility. Organ distribution in normal and nude mice. The simultaneous detection of Ig and theta on the surface of mouse lymphocytes permits the detection of four cell types (Ig-theta-, Ig-theta+, Ig+theta-, and Ig+theta+) which also have distinct electrophoretic profiles. All types are present in variable proportions in all lymphoid organs studied. Results obtained in congenitally athymic nude mice and in T-deprived mice define a new type of lymphocyte, the Ig-theta+weak. This cell is non-recirculating, Ig-, with a low density of theta and a slow electrophoretic mobility. It is a candidate for a T-committed prethymic cell.


Cell-mediated immunity to Moloney sarcoma virus in mice. II. Analysis of antigenic specificities involved in T lymphocyte-mediated in vivo rejection of murine sarcoma virus-induced tumors. Sublethally irradiated BALB/c mice innoculated with Moloney sarcoma virus (MSVm) develop progressively growing tumors and die within 30 days of virus innoculation. These animals can be protected from tumor progression (and death) by innoculation of small numbers of MSV-immune T lymphocytes from MSV-M innoculated (but unirradiated) animals. T lymphocytes in these donor animals have been shown to express immunity to a variety of viral and virally-induced antigens. We have investigated whether immunity to any one of these antigens was critically important in leading to protection of the irradiated animals by sensitizing normal T lymphocytes in vitro to different viral antigens and examining the ability of these sensitized cells to protect the irradiated recipients. Data is presented to show that cells sensitized in vitro to MSV-transformed fibroblasts, and to purified antigens with group specificity, but not to viral envelope antigens, or whole virus, are capable of protecting the irradiated MS innoculated animals.


The failure to show a necessary role for C3 in the in vitro antibody response. The in vitro antibody response of mouse spleen cells to TNP coupled to both T-dependent and T-independent carriers as well as to sheep erythrocytes has been studied to investigate the possible role of complement activation in the induction of antibody formation. The following has been found. (1) In vitro responses of both IgM and IgG can be obtained to both T-dependent and T-independent antigens in serum-free media, although they are smaller than those found in serum-containing media. This shows that no exogenous source of complement is necessary for in vitro antibody formation by spleen cells. (2) Similarly, normal antibody responses are obtained if the cultures are grown in human serum depleted of C3b-inactivator, which contains high concentrations of C3b. (3) In the presence of antibody to mouse C3 the response to the T-independent antigen is reduced, the IgM responses being more affected than the IgG. However, purified F(ab')2 anti-C3 has no inhibitory effect and it therefore seems likely that it is the formation of intact immune complexes containing Fc rather than the interference with C3 function that is responsible for the inhibition seen. (4) The conventionally purified anti-complementary factor from cobra venom has no effect on the antibody response in serum-free culture or when human or fetal calf sera are used. In no experiment was any potentiation of T-dependent responses observed. However, the presence of quite small concentrations (2%) of cobra veno


Differing mechanisms of tolerance and desensitization to dinitrochlorobenzene in guinea pigs. In the present paper the mechanisms of tolerance and desensitization to dinitrochlorobenzene (DNCB) contact sensitivity in guinea pigs were investigated using the methods of adoptive sensitization of tolerant and normal syngeneic recipients and cyclophosphamide-treatment of tolerant animals known to selectively inactivate suppressor lymphocytes. It was shown that desensitization of presensitized animals is caused by the direct effect of the intravenously injected hapten on the effector cells in the peripheral compartment. The immediate onset of unresponsiveness and its very short duration almost exclude the possible involvement of enhancing antibodies or suppressor cells. In the case of tolerance induced by pretreatment with dinitrobenzenesulfonic acid, suppressor cell activity is enhanced, preventing normal specific immunocompetent cells from recognizing the antigen and/or proliferating in the draining lymph nodes. Whether suppressor lymphocytes are of the B or T type is not yet known.


The failure to show a necessary role for C3 in the in vitro antibody response. The in vitro antibody response of mouse spleen cells to TNP coupled to both T-dependent and T-independent carriers as well as to sheep erythrocytes has been studied to investigate the possible role of complement activation in the induction of antibody formation. The following has been found. (1) In vitro responses of both IgM and IgG can be obtained to both T-dependent and T-independent antigens in serum-free media, although they are smaller than those found in serum-containing media. This shows that no exogenous source of complement is necessary for in vitro antibody formation by spleen cells. (2) Similarly, normal antibody responses are obtained if the cultures are grown in human serum depleted of C3b-inactivator, which contains high concentrations of C3b. (3) In the presence of antibody to mouse C3 the response to the T-independent antigen is reduced, the IgM responses being more affected than the IgG. However, purified F(ab')2 anti-C3 has no inhibitory effect and it therefore seems likely that it is the formation of intact immune complexes containing Fc rather than the interference with C3 function that is responsible for the inhibition seen. (4) The conventionally purified anti-complementary factor from cobra venom has no effect on the antibody response in serum-free culture or when human or fetal calf sera are used. In no experiment was any potentiation of T-dependent responses observed. However, the presence of quite small concentrations (2%) of cobra venom factor (CVF)-treated normal mouse serum produced a profound inhibition of the antibody responses, affecting IgG responses more than IgM, and being T-dependent more than T-independent, but no part of the response being unaffected. However, CVF heated to 70 degrees C which destroys the anticomplementary activity but not the contaminating phospholipase A was equally effective. More highly purified CVF, on the other hand, was not. These findings strongly suggest that the inhibition observed is due to residual phospholipase A reacting with lecithin in the mouse serum rather than to any anticomplementary effect. (5) Spleen cells taken from mice-treated with CVF in vivo were also unable to give a full IgG response to the T-dependent antigens if challenged 24 h after CVF treatment. However, after 48 h no inhibition was observed. It therefore seems to us that it is unlikely that C3 plays any real part in the in vitro antibody response. Our data--like the occurrence of genetically C3-deficient humans with apparently normal antibody responses--sould seem to preclude any necessary role for C3 in antibody production.


Immunological memory function of the T and B cell types: distribution over mouse spleen and lymph nodes. Spleens from LAF1 mice injected intravenously with sheep erythrocytes (SE) are relatively rich in memory T cells early in the immune response (1 to 3 days) and rich in memory B cells as the response progresses (2 weeks or more). Marked cooperation for the secondary immune response in vitro was obtained by combining 10(6) spleen cells from LAF1 mice, taken 2 days after intravenous priming with SE, with 10(7) spleen cells from day 14 primed mice. The results indicate relative deficiencies in the spleen for B memory cells on days 1 to 2 and for T memory cells on day 14 after priming. Day -14, but not day -2, immune lymph node (LN) cells could replace the day -2 spleen cells (anti-Thy 1.2 sensitive) in the in vitro cooperation with day -14 immune spleen cells. Immune spleen cells taken 4 to 7 days after priming contain more equivalent numbers of B and T memory cells, but 10 to 7 days after transfer of such immune spleen cells without SE into irradiated recipients the T memory cells were again more prominent in lymph node and the B memory cells in spleen as shown by in vitro cooperation studies. These results suggest that during the second week after intravenous injection of SE relatively more T than B memory cells migrate from spleen to lymph node, resulting in an imbalance in the splenic memory cell population favoring B memory cell function.


Lack of requirement for thymocytes for efficient antibody formation to trinitrophenylated mouse red cells in mice: role for thymocytes in suppression of the immune response. The requirement of thymus-derived cells for the immune response to trinitrophenylated syngeneic mouse red cells (TNP-MRC) was investigated. In three sets of experiments the following results were obtained: a) irradiated mice which were reconstituted with bone marrow cells alone showed a better anti-TNP response after injection with TNP-MRC than those reconstituted with both bone marrow cells and thymocytes. b) Anti-thymocyte serum augmented the mouse anti-TNP response to TNP-MRC. c) Nude thymusless mice showed a better anti-TNP response to TNP-MRC than their normal littermate controls. These results indicate that the anti-TNP response of mice to TNP-MRC does not require thymus-derived helper cells. Moreover, thymus-derived cells have a suppressive effect on the anti-TNP response.


Turnover of radioiodinated and of leucine-labeled immunoglobulin M in murine splenic lymphocytes. Lactoperoxidase-catalyzed radioiodination of cell surface proteins and biosynthetic incorporation of tritiated leucine were used to study the size and turnover of cell-associated immunoglobulins in splenic lymphocytes of different size from BALB/c, C3H, and nu/nu mice. Small spleen cells, separated from large cells by velocity sedimentation, showed a single slow rate of turnover (t1/2 = 10-30 h) of surface radioiodinated or leucine-labeled IgM released as the monomeric 7-8 S subunit of IgM. Mitogen-activated large B cells and nonstimulated large spleen cells, separated by velocity sedimentation, released leucine-labeled IgM in an initial rapid phase (t1/2 = 2-4 h) as 19 S IgM, and later released 7-8 S IgM slowly (t1/2 = 10-30 h). The iodination reaction changed neither the biphasic mode of turnover, nor, even at higher concentration of H2O2, the size of the actively secreted 19 S IgM. Radioiodination of mitogen-activated large cells or of unstimulated large cells labeled only the slowly released 7-8 S IgM, not the actively secreted 19 S IgM. Large cells of nonstimulated spleen, however, released radioiodinated as well as leucine-labeled IgM also rapidly (t1/2 = 1-3 h). These rapidly released IgM molecules were found to be 7-8 S IgM subunits. Our results indicate that these turnover rate and size differences of radioiodinated and of leucine-labeled IgM released from murine spleen cells are in part resulting from the differences in the methodology of labeling and are in other parts due to different small and large splenic lymphocytes releasing IgM of different size at different rates.


Histocompatibility antigens acting as helper determinants for tumor-associated antigens of murine lymphosarcoma. The mechanism of effectiveness of allogeneic versus syngeneic immunization in mounting a humoral specific immune response against tumor-associated antigens of murine urethan-induced thymic lymphosarcomas was studied. C57BL mice immunized with untreated or blocked C3Hf lymphoma cells yielded sera with complement-dependent cytotoxicity for C57BL lymphosarcomas, whereas they did not respond to repeated injections of blocked syngeneic lymphoma cells, indicating a helper activity of alien alloantigens in the reflex of tumor antigens. Thymectomized, lethally-irradiated and T, but not B-reconstituted mice produced active sera, suggesting a T-dependent response. The anti-tumor activity of the serum ran parallel with the anti-histocompatibility one. The tumor-associated and the alien histocompatibility determinants had to be present on the same cell membrane. In addition, active sera were obtained when immunizing syngeneic lymphoma cell inocula were followed by allogeneic normal immunocompetent cells. It is suggested that either a helper mechanism with B and T cells cooperating in recognizing haptenic tumor-associated and carrier histocompatibility antigens or a graft-versus-host reaction determining an abnormal induction of the immune system is needed in order to detect the weak tumor-associated antigens of urethan-induced lymphomas.


Target cell-dependent T cell-mediated lysis of vaccinia virus-infected cells. Vaccinia virus specific cytotoxicity against infected target cells was observed in vitro. Spleen lymphocytes from normal and immunized mice of the inbred strains C3H and DBA/2 were incubated with vaccinia virus-infected and non-infected 51 Cr-labeled mastocytoma P-815-X2 cells and L-929 fibroblasts, which were used as targets. Cytotoxic lymphocytes could be isolated from the mice as early as 2 days after infection with vaccinia virus. The highest cytotoxic effect was obtained with lymphocytes taken 6 days after infection. The degree of lysis was correlated with the ratio of immune lymphocytes to target cells. Specific blocking of target cell lysis resulted after addition of anti-vaccinia antibody from different sources. The effector cells could be characterized as T cells by elimination of macrophages and B cells. Target cell killing was only possible in a syngeneic system; allogeneic infected target cells were not lysed significantly.


Subclasses of T cells with different sensitivities to cytotoxic antibody in the presence of anesthetics. When cytotoxic titrations were made of rabbit anti-mouse brain and anti-Thy-1.2 sera against mouse lymphoid cells in the presence of sodium barbital or xylocaine, it was observed that a proportion of T cells were much more sensitive (50 x) than the remainder. This effect could be detected with ethidium or fluorescein diacetate as indicators of cell death, but not with trypan blue or 51Cr. The highly sensitive cells were found in all lymphoid tissues, being most numerous in spleen and least in lymph nodes.


p-Azobenzenearsonate-L-tyrosine-mediated helper function in immune responses of guinea pigs and rats. A small bifunctional antigen (4-hydroxy-5-iodo-3-nitrophenyl)acetyl-epsilon-aminocaproyl-L-tyrosine-azobenzene-p-arsonate [NIP-cap-TYR(ABA)] was found to induce fair humoral antibody formation against NIP-cap but very little anti-ABA-TYR. This was observed in rats and guinea pigs. Prior immunization with ABA-TYR, either as such or coupled to dodecanoylated bovine serum albumin (lipid-BSA), primed rats for an enhanced anti-NIP response to NIP-cap-TYR(ABA). An attempt to encourage rats to produce anti-ABA-TYR in response to the bifunctional antigen by priming them with NIP-cap-lipid-BSA failed. Priming with ABA-TYR was dose-dependent. An injection of 1.5-15 nanomoles per rat primed for an increased production of anti-NIP while 150 nanomoles did not. Adult thymectomized x-irradiated rats had a poor anti-NIP response to the bifunctional antigen if they were reconstituted with T-enriched lymphoid cells from control mice, but a good response if reconstituted with similar cells from ABA-TYR-primed syngeneic rats.


Nonspecific acid esterase activity: a criterion for differentiation of T and B lymphocytes in mouse lymph nodes. A modified technique was used to demonstrate lymphocytic acid alpha-naphthyl acetate esterase activity in frozen sections of mouse lymphoid tissue or smears. Positive, dot-like reaction products were noticed in more than 94% of the lymphocytes located in the diffuse cortical ("paracortical", "thymus-dependent") area of mesenteric lymph nodes of young adult ICR mice. Almost identical values were found for lymphocytes in the cisterna chyli. In contrast, the follicular cortex which is predominantly occupied by B cells, contained less than 7% esterase-positive lymphocytes. In vitro, cytotoxic anti-theta serum destroyed the vast majority of esterase positive lymphocytes while esterase-negative lymphocytes were resistent to this treatment. In mesenteric nodes of nude BALB/c (nu/nu) mice, follicular cortex and paracortex together contained approximately 16 times less esterase-positive lymphocytes than in the immunologically competent hybrid BALB/c (nu/+) animals. These findings indicate that non-specific acid esterase activity may serve as a criterion to differentiate peripheral T and B lymphocytes in lymph node sections and smears of mice by light microscopy. Possible implications of this enzyme activity in thymus-derived lymphocytes are discussed.


H-2-linked genetic control of antibody response to soluble calf skin collagen in mice. An autosomal dominant immune response gene could be demonstrated in congenic resistant strains of mice which is linked to the H-2 locus and controls the antibody response to soluble calf collagen. High responsiveness was associated with the H-2 alleles, b and f, low responsiveness with the H-2 alleles, d, k, m and r. Studies with calf procollagen, which contains an additional carrier moiety, indicated that these genetic differences might be expressed at the level of T cells.


Regulation of the immune response by subclasses of T lymphocytes. I. Interactions between pre-killer T cells and regulatory T cells obtained from peripheral lymphoid tissues of mice. The generation of killer cells to alloantigens in vitro depends upon an interaction between two subclasses of peripheral T cells: "pre-killer" T cells and "regulatory" T cells. The pre-killer T cell, found in highest concentrations in peripheral lymph nodes of mice, is sensitive in vivo to the administration of small doses of anti-thymocyte serum (ATS) and is depleted in vitro by treatment with (a) anti-Thy-1.2 + C, (b) low doses (600 r) of x-irradiation. Specificity of pre-killer cells is supported by adsorption of pre-killer activity on appropriate allogeneic monolayers and by the specific absence of this activity from lymphoid tissues of mice rendered tolerant in neonatal life. Although T cells remaining in the spleen 2 days after a small dose of ATS (SPA-T) did not generate killer cells, this subset exerted substantial regulatory effects upon the generation of killer activity by pre-killer T cells. The addition of this population of regulatory T cells to small numbers of lymph node T cells (LN-T) resulted in substantial enhancement in the generation of killer cells from the LN-T pre-killers. The addition of SPA-T to larger numbers of LN-T (which produce strong responses alone), failed to enhance the response, and in some cases resulted in significant suppression which could not easily be accounted for by alteration in cell numbers in culture. Although amplifier activity was relatively radioresistant, "suppressor" activity was sensitive to relatively small doses of irradiation. The significance of this T-T interaction is discussed.


Regulation of the immune response by subclasses of T lymphocytes. II. The effect of adult thymectomy upon humoral and cellular responses in mice. The immunologic consequences resulting from thymectomy in adult life were investigated. Primary humoral responses were not diminished shortly after adult thymectomy, as judged by responses of intact thymectomized mice as well as by the ability of spleen cells from such mice to transfer primary responses. However, secondary humoral responses were substantially reduced in irradiated recipients of spleen cells from primed adult-thymectomized mice, suggesting that the thymus is required in adult life to maintain a population of cells important in the generation of immunologic helper memory. By contrast, small doses of anti-thymocyte serum, which primarily affect recirulating T cells, abrogated the primary humoral response, but allowed the subsequent development of immune memory. Using a technique permitting in vitro sensitization of purified T cells to alloantigens, it was shown that adult thymectomy increases the ability of T cells to generate primary cytotoxic responses, but had little effect upon the development of cytotoxic T memory activity. These experiments suggest that in adult life the thymus maintains a regulatory population of T cells in peripheral tissues which suppress early T cell differentiation to cytotoxic effector cells and potentiates the development of immune memory.


Antibody response to phosphorylcholine in vitro. I. Studies on the frequency of precursor cells, average clone size and cellular cooperation. The anti-phosphorylcholine (PC) antibodies synthesized by BALB/c spleen cells in microcultures upon immunization with heat-killed vaccine of Pneumococci R36A (Pn) are directed exclusively to the PC epitope. These antibodies are of very restricted avidity and 88% of the responding clones express the idiotype characteristic of the TEPC-15 PC-binding myeloma. This idiotypic restriction appears to be due to the absence of clones capable of expressing other idiotypes, rather than to "clonal dominance". The estimated frequency of precursor cells for the PC epitope is 1 X 10(-5) to 2.5 X 10(-5). These precursors give rise to clones with an average size of 9 plaque-forming cells. When the logarithm of the number of negative wells was plotted against the number of spleen cells/well, the fraction of nonresponding cultures decreased exponentially as the number of spleen cells was increased. This indicated that only one cell type was limiting in our assay, presumably a B cell. Furthermore, treatment of spleen cells with AKR anti serum completely abolished the response to sheep red cells without affecting the response to PC. It is concluded that PC is a T cell-independent antigen. Of interest was the finding that PC requires adherent (A) cells and this is a particular characteristic of PC, since most T cell-independent antigens have been found not to require A cells. Reasons for the possible homogeneity of the response to PC are also discussed.


Recruitment of effector lymphocytes by initiator lymphocytes: role of a trypsin-sensitive membrane component. In earlier studies we found that initiator T lymphocytes sensitized in vitro against fibroblasts were able to recruit immunospecific effector T lymphocytes in vivo in syngeneic mice. In the present study, to investigate the possible function in effector cell recruitment of sensitizing antigens passively adsorbed onto the initiator cell membrane, we treated the initiator lymphocytes with trypsin. We found that trypsin inhibited recruitment. However, the initiator cells spontaneously recovered their recruiting ability after incubation in vitro for 4 h in the absence of contact with sensitizing antigens. Recovery could be blocked by cycloheximide. Hence, it appears that recruitment depends on the intrinsic metabolic function of the initiator lymphocytes and is not due merely to passively-adsorbed antigen carried over into the recipient mouse.


Immunoglobulin turnover in B lymphocyte subpopulations. "In vitro" turnover of leucine-labeled and of radioiodinated IgM has been studied with cells from various lymphoid organs of nude mice, i.e. lymph nodes, thoracic duct, spleen and bone marrow, as well as with subpopulations of B cells from spleen and bone marrow separated by free flow electrophoresis. Three types of IgM-producing lymphocytes could be distinguished by their turnover rates of IgM, by the size of the released IgM and by the capacity of the IgM molecules to be labeled by the lactoperoxidase-catalyzed radioiodination reaction and/or by incorporation of radioactive leucine. Type I cells release 7-8 S IgM rapidly (t1/2 = 1-3h); the released IGM is leucine-labeled and radioiodinated. Type II cells release 7-8 S IgM slowly (t1/2 =10-30); the released IgM is leucine labeled and radioiodinated. Type III cells release 19 S IgM rapidly (t1/2 =2-4 h); the released IgM is leucine labeled, but not radioiodinated. Lymph nodes and thoracic duct contain predominantly type II cells, bone marrow contains type I and II cells, spleen contains type I,II and III cells. It is suggested that type III cells are Ig-secreting plaque-forming plasma cells, type II cells are small, resting "memory" B cells, and type I cells may be newly formed antigen-inexperienced B cells.


5-Hydroxytryptophan effect on the development of the immune response: IgM and IgG antibodies and rosette formation in primary and secondary responses. In animals immunized with bovine serum albumin, 5-hydroxytryptophan prolonged the latent period of the IgM and IgG primary responses, decreased response intensity, delayed the response peak and suppressed IgG immunological memory. In 5-hydroxytryptophan-treated mice, the number of rosette-forming cells (RFC) in the lymph node and spleen decreased during the primary and secondary responses. This effect was due to the decreased number of IgG RFC and to the later involvement of IgM RFC in the immune response. The absence of the secondary response was related to unprimed IgG memory cells. 5-hydroxytryptophan does not inhibit the primary and secondary responses after the connections between the hypothalamus and pituitary have been disrupted. The participation of the n.raphe-hypothalaminc-pituitary system in immuno-regulation and the putative mechanism underlying serotonin effect on the immune response are discussed.


Hapten-induced B cell paralysis. II. Evidence for trivial mechanisms of tolerance. Injection into mice of a free reactive form of the hapten (4-hydroxy-3,5-dinitrophenyl)acetyl (NNP), induces a state of specific unresponsiveness to the hapten, upon its challenge with thymus-cependent and independent carriers. This unresponsiveness is maintained in vitro. Both induction and expression of the unresponsive state were found to be independent of T cells. Analysis of the mechanisms responsible for the B cell "tolerance" demonstrated that the major cause of unresponsiveness in this system is the blockade of specific surface receptors on B cells by the hapten, abolishing the focusing function of these receptors. "Tolerant" B cells, though they were unresponsive to the antigen, could be activated to anti-hapten antibody secretion by the B cell mitogen lipopolysaccharide (LPS), which does not require Ig-mediated binding. They also became fully responsive to the antigen NNP-LPS in specific thymus-independent responses after 24 hours of in vitro incubation in the absence of tolerogen, conditions which allow "deblocking" of Ig surface receptors, and, thus, restoration of the focusing function of these receptors. These results demonstrate that great care is required for a clearcut definition of B cell tolerance. In this example, the tolerant B cells were perfectly responsive to a competent ligand, and no indication of an altered triggering or effector processes was found. It appears that in this, as in many other cases of B cell tolerance, the systems or the animals are tolerant, whereas B cells maintain a resting nonactivated state and are fully responsive to the triggering signal. Thus, the existence of tolerance-inducing signals resulting in B cell unresponsiveness is questioned.


Immunological effects of IgT synthesized by theta-positive cell lines. The effects of immunoglobulin (Ig) purified by affinity chromatography from two cultured T cell lines on in vitro immune responses was studied. IgT purified from both EL-4 and WEHI-22 suppressed the thymus-dependent IgM response, but had no effect on the thymus-independent IgM response. In contrast, the IgG responses to both thymus-dependent and independent antigens were augmented at optimal concentrations of IgT. Ig of B cell origin--serum Ig, myeloma Ig, B cell surface Ig -- did not have any of these effects. Because of the similarity of the effects of IgT purified from the lymphomas with the effects of T cells and their IgT, it seems likely that the IgT obtained from EL-4 and WEHI-22 cell lines are analogues of normal IgT and thus should be useful in elucidating some of the chemical and biological properties of IgT.


A simple rosette assay for demonstration of complement receptor sites using complement-coated zymosan beads. In the present article we describe a simple rosette assay for detection of C3 receptor-bearing B lymphocytes with complement (C)-coated zymosan (Zy) beads. Zy coated with murine or human C bound to a distinct population of human and mouse lymphocytes as well as to the majority of lymphoblastoid cells of several human established cell lines. Rosette formation was also observed with human red blood cells, with human monocytes and neutrophils. Experiments with anti-immunoglobulin sera, with other B and T cell markers and with mouse thymocytes proved that the capacity to bind C-coated Zy is primarily a feature of B lymphocytes. The following findings suggested that C-coated Zy is bound via receptor sites for the activated components of C3: (a) Zy coated with C3-deficient serum failed to bind, (b) comparable percentages of various lymphoid and nonlymphoid cells formed rosettes with C-coated Zy as well as with antibody and C-coated sheep red blood cells, and (c) antibodies against human or murine C3 inhibited the binding of C-coated Zy.


Ontogeny of the antibody-forming cell line in mice. I. Kinetics of appearance of mature B cells. The development of functionally mature B cells in the liver and spleen of fetal and neonatal CBA mice has been followed. A mature B cell is defined as one giving rise to antibody-producing progeny within 2 days in culture. Very few such B cells are found during fetal life. After birth there is a rapid rise in the numbers reaching 1/3 of adult levels in the 7-day-old spleen.


Suppression of immune response to sheep red blood cells in mice treated with preparations of a tumor cell component and in tumor-bearing mice. The frequency of rosette-forming cells (RFC) in spleens of mice immunized with sheep red blood cells (SRBC) was significantly reduced by pretreatment with preparations obtained from tumor extracts. This effect was not produced by preparations obtained by similar methods from normal tissue extracts. Significant suppression of RFC frequency was also found in spleens of mice immunized with SRBC at late stages of tumor growth, as compared to that in immunized controls and in mice bearing tumors at early stages. The results suggest that at late stages, a tumor cell component is released by tumor cells which may be similar to the nondialyzable immune suppressive factor(s) separated by us from tumor extracts, factors found in ascites fluid of tumor-bearing mice and those known to be released from tumor cells in vitro.


Persisting T cells in rats tolerant of human serum albumin. The significance of tolerant and nonimmune T cells which preferentially restrict high affinity antibody synthesis. The adoptive response of primed rat thoracic duct lymphocytes ('TDL) following specific antigen challenge (soluble human serum albumin, s-HSA) was restricted when cells were transferred into syngeneic, adult (AS2 X AS)F1 hybrid recipients in comparison with irradiated hosts. This adoptive memory response was also inhibited in irradiated recipients by transferring nonimmune TDL along with 'TDL. Recirculating B cells (B-TDL) did not inhibit the 'TDL response, indicating that the adoptive secondary response was regulated by T cells. Antibody synthesis was preferentially restricted in the high affinity memory cell precursor population, demonstrating a role for T cells in regulating the maturation of antibody affinity. The adoptive memory response was liberated from this T regulatory effect in adult recipients when hosts were challenged with the alum-precipitated adjuvant form (HSA-adj) rather than the soluble form of HSA. Since the adoptive memory response was sensitive to the presence or absence of T cells, this experimental model was used to determine whether or not T cells were eliminated from HSA-tolerant rats. Antibody synthesis by 'TDL was reduced approximately 10-fold compared with controls when transferred into tolerant recipients and challenged with either s-HSA or HSA-adj; a similar reduction was not observed by substituting bovine serum albumin (BSA) 'TDL and challenging with s-BSA. The tolerance-induced inhibition of HSA 'TDL was destroyed by irradiation and TDL from HSA-tolerant donors were more effective than normal nonimmune TDL in reducing the adoptive HSA 'TDL response. HSA-tolerant TDL did not inhibit the BSA 'TDL response significantly. The results indicate that T cells are not eliminated by tolerance induction in this model and after interaction with tolerogen may exert an active (or competitive) role in restricting antibody synthesis by high affinity B memory cell precursors. However, the fact that tolerant T cells are not able to prevent a primary response suggests that unresponsiveness to HSA in the T compartment represents a functional deficiency and not an active suppression at this level. Nevertheless, the presence of these tolerant cells probably accounts for the failure of antibody affinity to mature in partially tolerant rats.


X-ray enhancement of splenic rosette-forming cells in nonimmune mice. The formation of rosettes between spleen cells from nonimmune mice and sheep red blood cells (SRBC) has been investigated. The number of splenic rosette-forming cells (RFC) in the intact spleen was very low but greatly increased after sublethal whole body x-irradiation. These cells were inhibited by pretreatment of the cells with AKR anti-C3H- antigen antiserum, in the presence of fresh guinea pig serum, but not with rabbit anti-mouse immunoglobulin antiserum. The splenic RFC also increased after cortisone acetate administration into mice. These results suggest that the RFC are a T cell sub-population having x-ray resistant and cortisone-insensitive characteristics. The x-irradiation of mice in vivo selectively increased the number of splenic RFC in consequence of reduction in the number of cells having non-T cell characteristics. These RFC are presumably antigen-specific sheep red blood cell rosettes, a quite different situation as in the case of human T lymphocyte rosette formation.


Analysis of the role of different cell types in the genetic regulation of antibody production to the thymus-independent synthetic polypeptide poly (DTyr, DGlu)-poly (DPro)--poly (DLys). The immune response potential of mice to the thymus-independent synthetic polypeptide poly (DTyr, DGlu)-poly(DPro)--poly(DLys)[D(T,G)-Pro--L] is genetically regulated. The defect in the ability of low responder mice to mount an immune response to this antigen appears to be expressed in their B cell population since the presence of thymocytes, or addition of "educated T cells" or supernatant of T cells after stimulation with the antigen neither enhanced, nor suppressed the level of antibodies produced in both low and high responder mice. Low responsiveness could not be enhanced either by stimulation of macrophages or by injection of poly(A) - poly(U) in contrast to the significant effect of these agents on low responses to the thymus-dependent poly(LTyr, LGlu)-poly(LPro)--poly(LLys) [L(T,G)-Pro--L]. These results suggest that macrophages do not participate in the limiting step, or are not involved at all, in antibody production towards the thymus-independent polypeptide. The antibodies produced in response to D(T,G)-Pro--L were found to be mainly of the 7 S class. T cells are not required for the production of mercaptoethanol resistant antibodies to this immunogen since they were found in intact mice as well as in T cell depleted animals.


Role of epitope density in the induction of immunity and tolerance with thymus-independent antigens. II. Studies with 2,4-dinitrophenyl conjugates in vivo. The immunogenicity and tolerogenicity of 2,4-dinitrophenyl (DNP) conjugates of levan, type 3 pneumococcal polysaccharide (SIII), dextran B512 and dextran B 1299 with different levels of substitution were assessed in vivo. It was confirmed that, as in vitro, conjugates with low epitope density are solely immunogenic for Bmu cells, whereas higher density conjugates are both immunogenic and tolerogenic according to dosage. The maximal anti-DNP plaque-forming cell responses attainable varied by 2 log10 according to the carrier, being greatest with dextrans and least with SIII. The effect of the carrier on immunogenicity was not attributable to B cells responding to it, as concurrent tolerization by either free polysaccharide or the conjugate itself was without effect. Optimal immunization was succeeded by varying degrees of responsiveness ("exhaustion") which was greatest with the more immunogenic conjugates. In contrast, reduction of tolerizing dose by cyclophosphamide suppression was only a feature of more highly substituted conjugates.


The effect of the thymus-independent antigens, collagen and synthetic collagen-like polypeptide, on the requirement of cell cooperation in the immune response to thymus-dependent antigens. The effect of thymus-independent antigens on the need for cell cooperation in the immune response to thymus-dependent antigens was investigated. Irradiated recipient mice transplanted with either bone marrow cells or a mixture of bone marrow and thymus cells, were immunized with the thymus-independent antigen (Pro-Gly-Pro)n covalently conjugated to the thymus-dependent ovalbumin, or with a mixture of (Pro-Gly-Pro)n and ovalbumin. In both cases an effective response towards ovalbumin was observed in the absence of thymus cells as was found for the thymus-independent (Pro-Gly-Pro)n. The same effect on ovalbumin was demonstrated when a mixture of the thymus-independent collagen and ovalbumin was used for immunization. On the other hand, when irradiated reconstituted mice were immunized with a mixture of ovalbumin and the thymus-dependent gelatin, which is the denatured product of collagen, cell-to-cell cooperation was required for an immune response to both immunogens. The effect of (Pro-Gly-Pro)n and collagen on the response to the thymus-dependent ovalbumin in vivo was observed in in vitro experiments using sheep red blood cells (SRBC) as the immunogen as well. In the presence of reduced and carboxymethylated (RCM) Ascaris collagen and (Pro-Gly-Pro)n, nude spleen cells could produce significant numbers of plaque-forming cells towards SRBC. Thus, (Pro-Gly-Pro)n and collagen can deliver the signal required to stimulate B cells to produce antibody towards thymus-dependent antigens in the absence of T cells. In contrast to the results with (Pro-Gly-Pro)n and collagen, the thymus-independent synthetic polypeptide poly(DTyr, DGlu-)-poly(DPro)--poly(DLys) did not affect the requirement for cell cooperation of the thymus-dependent immunogens, ovalbumin and SRCB. It thus appears that the ability to substitute for T cells for antibody production towards thymus-dependent immunogens is not a general characteristic of thymus-independent antigens.


Expression of a receptor for IgM by human T cells in vitro. The capacity of human peripheral blood lymphocytes (PBL) to bind antigen-IgG antibody (EA(IgG)) and antigen-IgM antibody (EA(IgM)) complexes was investigated using a rosette technique with ox erythrocytes coated with rabbit IgG or IgM antibody. It was found that while EA(IgG)-rosette-forming cells (RFC) were detected on PBL freshly drawn from normal individuals, EA(IgM)-RFC were present only in suspensions kept in culture for 24 h in mediá supplemented with sera containing very low or no amounts of IgM. Experiments of simultaneous detection of EA(IgG)-RFC or EA(IgM)-RFC and other membrane markers for human T or B cells together with experiments on purified T or B cell populations indicated that EA(IgG)-RFC were formed by both T and B cells, while T cells only were capable of EA(IgM) rosette formation. The specificity of the receptors for IgG and IgM was determined by studying the inhibitory capacity of purified human IgM and IgG in the rosette assay. The receptor for IgG was inhibited by IgG and not by IgM, while the reverse was true for the receptor for IgM.


Studies on the subpopulation and function of peripheral lymphocytes, and inhibitor to PHA stimulation existing in the serum of patients with liver disease. In our recent report we observed the abnormal response to PHA stimulation in the peripheral lymphocytes of patients with chronic liver diseases. The present report concerns extensive studies of this earlier report; analysis of the subpopulation and function of peripheral lymphocytes in patients with liver diseases. PHA responsiveness in acute hepatitis was reduced in the acute phase and improved during the convalescent stage. In chronic liver disease, half of the patients showed a decrease in T-cell population as well as a reduced response to PHA stimulation. These phenomena were observed more frequently in HBs-antigen positive patients than in HBs-antigen negative cases. Furthermore, we found the existence of an inhibitor to PHA response in the serum of 8 cases among 19 patients with various liver diseases. In patients with such a serum inhibitor, the reactivity of the lymphocyte itself to PHA was considerably higher but the T-cell population was much more decreased than that of inhibitor negative cases.


Case report: a bleeding Meckel's diverticulum diagnosed by selective arteriography. The diagnosis of a bleeding Meckel's diverticulum was made by angiography in a 71 year old man who presented with melena and hemorrhagic shock. Emergent angiographic examination revealed an extravasation of contrast media from the ileal artery, which suggested a bleeding from the Meckel's diverticulum. The case illustrates the utility of selective angiography in case of gastrointestinal hemorrhage, where the lesion is beyond the range endoscopy.


Comparative, immunological studies on lymphangiectasia of the small intestine revealed in protein losing gastroenteropathy and Behçet's disease. Lymphangiectasia of the small intestine was demonstrated in 9 to 18 cases with protein losing gastroenteropathy and in 10 of 26 cases with Behçhet's disease. Protein losing gastroenteropathy was compared to Behçet's disease in view of immunological aspects. Immunoglobulin containing cells in the jejunal mucosa of protein losing gastroenteropathy were decreased, whereas Behçet's disease had normal or increased distribution. This suggested that suggested that immunoglobulin may be lost into the intestinal lumen or that production of those cells may be inhibited in protein losing gastroenteropathy. Decreased DNCB skin reaction and impaired blastoid transformation of peripheral lymphocytes in both of two diseases suggested that there should be immunological defect in those diseases. However, in Behçet's disease, investigation of the thymus disclosed hyperplasia and the presence of lymphoid follicle which does not appear in normal state. In conclusion, protein losing gastroenteropathy is in immunological deficiency state due to the congenital or acquired disorders of lymphatics which implies abnormal protein loss, decreased immunoglobulin containing cells in the jejunal mucosa and decreased sensitivity. On the contrary, in Behçet's disease hyperimmune state is present based on thymic hyperplasia.


How to detect and treat arrhythmias in chronic lung disease. Investigators suspect that arrhythmias are behind the high incidence of sudden death in patients with severe chronic bronchitis and emphysema. Continuous electrocardiographic monitoring may help boost survival statistics, since it has repeatedly shown its ability to detect irregularities that standard ECGs miss. So-called benign arrhythmias, such as atrial fibrillation, frequent ventricular premature contractions, and atrial flutter, are anything but harmless. In patients whose heart and lung function is already impaired by severe airway obstruction, these rhythm disorders can rapidly progress to a fatal cardiac arrest. Deciding how to treat the arrhythmia depends on the type of disturbance and the patient's clinical status. Direct-current shock or antiarrhythmic drugs can turn the tide in a desperately ill patient with acute respiratory failure. Correction of coexisting metabolic and ventilatory disorders often is crucial to successful therapy.


Ischaemic colitis in the experimental animal. I. Comparison of the effects of acute and subacute vascular occlusion. Acute ligation of the common colic and caudal mesenteric artery in the dog led to ischaemic colitis, whereas the acute ligation of the common colic artery and gradual occlusion of the caudal mesenteric artery by an ameroid plastic device did not lead to ischaemic disease. These findings suggest that the collateral blood vessels do not open up rapidly enough to prevent ischaemic change when the two major arteries are acutely ligated.


Peroral small-intestinal biopsy: experience with the hydraulic multiple biopsy instrument in routine clinical practice. Experience of the peroral, hydraulic, multiple, small-bowel biopsy instrument is recorded and compared with reported experience of other peroral biopsy instruments. It is concluded that, in routine clinical practice, there is no particular danger associated with this instrument despite warnings to the contrary. Furthermore, biopsies are obtained at least as quickly as with other instruments and with great reliability. Since this instrument also enables multiple, precisely located biopsies to be taken from various levels of the small intestine, it could be considered the instrument of choice for peroral jejunal biopsy.


Radioimmunoassay of secretin in plasma.

Radioimmunoassay of secretin in plasma. A sensitive, specific and reproducible radioimmunoassay for secretin is described. Antibodies were readily produced against low microgram quantities of synthetic secretin. The secretin antibodies did not cross-react with the structurally similar G.I.P., V.I.P., or glucagon. Synthetic secretin was iodinated using Chloramine "T" and purified by a two-state procedure incorporating gel filtration and radient elution from a cation exchange column. Plasma samples were found to produce variable interference in the assay necessitating the incorporation of secretin-free "blands" for each patient's plasma. Production of secretin-free plasma was by incubation of plasma samples at 37 degrees C for 96 hours. The sensitivity of the assay was 12.5-25 pg/ml. Normal fasting secretin levels were 21 +/- S.E. 7 pg/ml. A mean rise in plasma secretin to 220 pg/ml was observed after intraduodenal acidification.


Genetic variants of serum alpha1-antitrypsin (Pi types) in Portuguese. The results of Pi typing on 330 Portuguese from the area of Lisbon are reported. We found six phenotypes and four alleles out of the 24 described in the literature. The allele PiM is the most frequent as in other populations, PiS shows a high frequency (0.1152), and PiF is absent, which agrees satisfactorily with former studies carried out in Spain. These results are compared with others and the entity of the Iberian population is evoked.


Clonal dominance and the preservation of clonal memory cells mediated by antigen-antibody. Selected B-cell clones and their well characterized monoclonal antibody products were used to analyse the role of antibody in clonal dominance and the regulation of memory cell supplies. The experimental design was to permit contact between spleen cells and antigen in vitro, and to administer antibody to DNP prior to or following cell transfers into irradiated recipients. The anti-hapten response was strongly suppressed by the clone's own antibody or higher affinity antibody administered on day 0. Antigen-antibody inhibited memory cell generation. The suppressive effect was temporary, and reversible with time and further antigen and the same clone could be induced to produce antibody again, analysed by isoelectric focusing. We were therefore not dealing with clonal deletion. Change in the source of clonal anti-hapten excluded possible effects of antibody to carrier protein or idiotypic determinants in this system. The timing of antibody administration indicates that clones already triggered in the first 4 days after antigen contact could not be suppressed by antibody. Passive antibody to DNP only suppressed when both B and T cells had been permitted contact with hapten-carrier protein. Alteration of the carrier protein enabled us to study the effect of antigen-antibody on B and T cells separately. B cells binding antigen and antibody to hapten were triggered more efficiently by fresh T cells recognizing the carrier protein than after antigen uptake alone. Antibody to DNP suppressed only when both B and T cells had taken up hapten-protein, suggesting that antigen-antibody acts centrally at the level of both B memory cells and T helper cells. This reversible antigen-antibody blockade appears to favour the preservation of a pool of long-lived memory cells rather than the priming of new clones developing from short lived precursor cells; clonal dominance ensues.


Endocardial lesions following coronary artery by-pass surgery. A series of post-operative deaths is presented in which coronary artery by-pass surgery had been performed at various ante-mortem intervals by the same surgical term in a single institution. When compared to an appropriate control population, these cases displayed a sequence of endocardial inflammation and progressive fibro-elastosis which suggested that injury had occurred during surgery. An assessment of the effects of this lesion and of possible etiologic factors is presented.


Lymphocyte responses to phytohaemagglutinin: age-related effects. Cell-mediated immunity is depressed in elderly individuals compared to young individuals, and lymphocytes from elderly individuals have been reported to have impaired lymphocyte responsiveness to stimulation by PHA after 4 days of culture. We have confirmed this observation. However, after 8 days of culture, the lymphocyte responses were greater in elderly normal individuals than in young normal individuals. Responses of lymphocytes from young individuals decreased with time from 4 to 8 days in culture, while there were increased responses with time when lymphocytes from elderly individuals were studied. When adherent cells from lymphocytes of young individuals were removed by  passage through protein-coated Degalan-bead columns, the lymphocyte responses to PHA were significantly increased at 8 days. Passage of lymphocytes from elderly individuals through coated Degalan bead columns did not alter the lymphocyte responses. Removal of macrophages from the mononuclear cells obtained from young individuals did not result in increased lymphocyte responses to PHA after 8 days in culture. Removal of adherent cells appeared to have the same effect regardless of the efficiency of removing B cells. The adherent cells removed by the protein coated columns, therefore, appear to be nonphagocytic mononuclear cells which are not B lymphocytes.


The enhancement of the haemolytic activity of the first component of complement by trasylol. Trasylol, a basic polypeptide extracted from bovine lung considerably enhances the complement activity of whole serum. The complement component influenced by Trasylol is C1, either in its activated form (C1) or in its precursor form (C1). An effect on the binding of C1 to the immunoglobulin molecule has been excluded and on becoming part of a cellular intermediate C1 has been shown to be insensitive to Trasylol. It seems that Trasylol modulates C1 in such a way as to render it more efficient haemolytically. An attempt has been made to explain these results by consideration of the relationship existing between complement and other biological systems, such as the fibrinolytic and kallikrein-kinin systems.


Autoreactivity developing spontaneously in cultured mouse spleen cells. III. Inhibition of anti-embryo cytotoxicity in male T lymphocytes by female non-T cells. Cultured female spleen cells develop less cytotoxicity to embryo-associated antigens than do cultured male spleen cells. This difference is less pronounced if the cells are treated before culture with a specific hetero-anti-B cell antiserum. Female non-T cells (especially from primiparous animals) can decrease the spontaneously developing T-cell cytotoxicity in male splenic T lymphocytes upon culture. By artificially manipulating the T cell: non-T cell ratio in the spleen population prior to culture, it seems one can alter the type of cytotoxic mechanism (to embryo-associated antigens) which develops in the cultures.