Release of prostaglandins in experimental immunecomplex endophthalmitis and phacoallergic uveitis. Prostaglandins-E were demonstrated in the aqueous humour, the anterior uvea, and the choroid of rabbits in which type III allergic reactions were produced by two different methods. In general the levels of prostaglandins were found to be higher in those animals in which the immune complexes were formed from autologous rather than heterologous tissue antigens.


Aniridia associated with microcornea and subluxated lenses. Four cases of aniridia associated with subluxated lenses and microcornea are presented. The triad occurred in both eyes of the 4 affected members in one Ndebele family (one of the South African Negro tribes). No other ocular or systemic defects were noted, and intelligence was normal. Chromosomal studies on both parents showed no abnormality, and gene marker studies failed to reveal any linkage between the disease locus and a wide range of polymorphic loci.


Posterior polymorphous dystrophy: a light and electron microscopic study. Microscopic study of a keratoplasty specimen in a case of posterior polymorphous dystrophy demonstrated via serial sections a multilaminar Descemet's membrane with focal fusiform nodular protrusions of connective tissue. The latter are thought to represent the polymorphous lesions of the posterior limiting layers of the cornea seen clinically. Ultrastructurally, fibroflast-like cells lined the posterior surface of the cornea. It is postulated that those reported cases of PPD in which epithelial-like transformation of the endothelial layer has been described may represent more advanced cases, in which corneal oedema is likely to be a prominant feature clinically. An evolving process of metaplasia rather than a static one might explain the variety of altered endothelial cells, fibroblasts, and epithelial cells which have been reported on the posterior cornea in the few PPD keratoplasty specimens studied to date.


Characterization of beta-hydroxybutyrate transport in rat erythrocytes and thymocytes. A method was developed for study of beta-hydroxybutyrate transport in erythrocytes and thymocytes. Critical to the method was a centrifugal separation of cells from medium which took advantage of beta-hydroxybutyrate transport's temperature dependence and inhibition by phloretin and methylisobutylxanthine, all of which are demonstrated in this work. These properies suggested mediated transport, as did saturation kinetics and inhibition by several agents including pyruvate and alpha-cyanocinnamate. Most conclusive in this regard was a 2-fold preference for D- over L-beta-hydroxybutyrate. Entry was not Na+ dependent. It was stimulated by substitution of SO2-4 for most of the Cl-. The equilibrium beta-hydroxybutyrate space was much higher than the Cl- space of thymocytes, suggesting that beta-hydroxybutyrate entry is not associated with net inward negative current and is not coupled to outward Cl- or inward K+ movement (assuming that K+ is at elecrochemical equilibrium). Coupling to H+ entry or OH- exit is compatible with the result. These findings are consistent with beta-hydroxybutyrate entry by the carboxylate transport site which has been studied extensively with pyruvate and lactate as permeants. The Cl-/HCO-3 exchange carrier did not appear to contribute significantly to beta-hydroxybutyrate transport.


Association of Tenebrio molitor L. alpha-amylase with two protein inhibitors--one monomeric, one dimeric--from wheat flour. Differential scanning calorimetric comparison of heat stabilities. Thermal stabilization resulting from protein . protein association between two protein inhibitors (coded as 0.19, a dimer, and 0.28, a monomer) from wheat flour and the alpha-amylase from Tenebrio molitor L. (yellow mealworm) larvae was investigated by differential scanning calorimetry (heating rate 10 degrees C/min). Thermograms (plots of heat flow vs. temperature) for the two inhibitors showed broad endothermic peaks with the same extrema (denaturation temperatures) at 93 degrees C, and equal, small enthalpies of denaturation (2 cal/g). The amylase produced a sharp endotherm at 70.5 degrees C, but a larger enthalpy change on denaturation (6 cal/g). The amylase . inhibitor complexes differed in thermal stability, but both showed significant stabilization relative to free enzyme. The complex formed with monomeric inhibitor 0.28 showed a higher denaturation temperature (85.0 degrees C) than that formed with dimeric inhibitor 0.19 (80.5 degrees C). This order of stabilization agrees with the relative affinities of the inhibitors for the amylase. These thermograms are consistent with previous results which indicated that 1 mol of amylase binds 1 mol of inhibitor 0.19.


Synthesis of antithrombin III and alpha-1-antitrypsin by the perfused rat liver. Livers isolated from control or turpentine-injected rats were perfused for 3 h with human red cells suspended in Krebs-Henseleit solution containing bovine serum albumin, dextran, glucose, heparin, cortisol, insulin, a mixture of 20 amino acids and [3H]leucine. Changes in the concentrations of antithrombin III and alpha-1-antitrypsin were evaluated by rocket immunoelectrophoresis using specific antisera, and incorporation of the 3H radioactivity into the total protein, albumin, antitrhombin III and alpha-1-antitrypsin in the perfusate was measured. The results indicate that both antithrombin III and alpha-1-antitrypsin are synthesized in the liver. Local inflammation induced in the liver donors moderately stimulated the synthesis of alpha-1-antitrypsin but it affected only marginally that of antithrombin III.


Colony-stimulating factors active on human bone marrow cells from a Yoshida sarcoma cell line. Colony-stimulating factors (CSF) active on both human and murine bone marrow colony-forming units in culture (CFU-C) were found in the conditioned medium of Yoshida sarcoma cells (line YSSF-212T), although the cells originated from rats. The CSF were inactivated by digestion with trypsin, alpha-chymotrypsin, and pronase. By chromatography on DEAE-cellulose, the CSF were separated into five subgroups with different capacities to stimulate mouse granulocyte and macrophage progenitors. CSF in these five peaks were eluted from an Ultrogel AcA 44 gel filtration column with apparent molecular weights of 22,000-25,000 daltons. CSF of nonhuman origin stimulating human CFU-C would be useful in hematologic studies of bone marrow cells.


Effect of neutral proteases from human granulocytes on colony forming cells in vitro. Marked inhibition of colony formation is observed after incubation of mouse and human bone marrow cells with the human granulocytic neutral proteases elastase and chymotrypsin as well as with pancreatic chymotrypsin. The corresponding enzymes inactivated with diisopropylfluorophosphate were almost inactive. Incubation of different colony inducing agents either resulted in no change or in an increase of their colony stimulating activity. The data suggest a direct proteolytic action of the proteases on colony forming cells which may alter receptor sites for colony stimulating activities.


Human placenta-conditioned medium for stimulation of human granulopoietic precursor cell (CFU-C) colony growth in vitro. Human placenta-conditioned medium (HPCM) has been reported to stimulate colony formation by human granulopoietic stem cells (CFU-C) in vitro. The present work was performed to further characterize this colony formation. The majority of HPCM batches tested stimulated colony growth equivalent to recombined human leukocyte feeder layers with optimal cellular composition. A broad plateau of the dose-response curve of HPCM was found. A linear correlation exists between the number of marrow cells plated and the number of colonies grown. Optimal duration of culture is between 9 and 11 days. Colonies are large and tend to be compact. Admixture of mature granulocytes does not affect the colony growth pattern under optimal culture conditions. These data document that HPCM is a suitable source of colony-stimulating activity for the routine assay of human CFU-C. Due to the constant colony stimulation, HPCM appears particularly valuable for longitudinal studies of human CFU-C.


[Influence of plasmaphoresis on total and T-lymphocyte count (author's transl)]. For more than one year 30 plasma donors were tested as to their influence on leukocytes, lymphocytes and SRBC-rosette-forming-T-lymphocytes. None of the donors, even under long-term observation and with more than 40 double plasmaphoreses per year, showed any significant changes in the above-mentioned data. The general level of lymphocytes and SRBC-rosette-forming is individually fixed, and there is a definite correlation between these two factors. This individual level is also uninfluenced by frequent plasmaphoreses.


Peduncular hemiplegia following removal of large cerebellopontine angle tumors: discussion of a mechanism of brain stem injury. Large cerebellopontine angle tumors distort and displace the brain stem. Two cases are reported in which ipsilateral hemiparesis ensued postoperatively. Compression of the contralateral cerebral peduncle against the free edge of the tentorium cerebelli appears to be the mechanism leading to this mesencephalic dysfunction. Review of the literature failed to reveal a discussion of this mechanism of brain stem injury. Extension of tumor through the tentorial hiatus appears to make the mesencephalon particularly vulnerable. Incision of the tentorium prior to removal of a tumor when it invades the incisural hiatus may have merit.


Stigmata of recent haemorrhage in diagnosis and prognosis of upper gastrointestinal bleeding. In 277 consecutive episodes of suspected upper gastrointestinal bleeding, lesions bearing stigmata of recent haemorrhage (stigmata) were found by endoscopy in 110 (47%) out of 233 patients who were judged to have bled; 78 (33%) had lesions without stigmata, and in 45 (19%) no lesion was seen. Results in 176 entirely unselected admissions for upper gastrointestinal bleeding were similar.Forty-eight chronic duodenal and 41 chronic gastric ulcers were identified by endoscopy. Stigmata were found in 27 (56%) and 33 (80%) of these cases respectively. Sixteen patients had multiple lesions, and in 12 (75%) the presence of stigmata permitted diagnosis of the source of the haemorrhage. Stigmata were more likely to be seen in cases of duodenal ulcer, Mallory-Weiss lesions, and oesophageal varices when endoscopy was performed within 12 hours of bleeding, but were as common in cases of gastric ulcer after longer intervals.In the absence of stigmata one out of 21 patients with duodenal ulcer had further haemorrhage and one other needed emergency surgery; no patient with gastric ulcer had further haemorrhage or needed emergency surgery. In contrast, when stigmata were present 15 of the 27 patients with duodenal ulcer (56%) had further haemorrhage and 17 (63%) needed emergency surgery; of the 33 patients with gastric ulcer, 10 (30%) had further haemorrhage and 15 (45%) required emergency surgery. Superficial mucosal lesions may have been the source of haemorrhage when an ulcer unmarked by stigmata was seen at endoscopy. Stigmata were superior to any other single factor or combination of factors in predicting rebleeding and the need for emergency surgery.


Studies on strain sensitivity of Trichomonas vaginalis to metronidazole. Ninety-four strains of Trichomonas vaginalis isolated from unselected patients in Vienna during the period from November 1975 to November 1975 were subjected to sensitivity tests against metronidazole in vitro and in vivo. Nearly 90% of the isolates examined showed in vitro growth inhibition at concentrations of 0.4 to 1.6 microgram metronidazole/ml and only about 10% at concentrations of less than 0.4 microgram/ml. All isolates showed some inhibition of growth after incubation for 48 hours with 3.2 microgram metroidazole/ml. Tests on treatment carried out on experimentally infected mice revealed that all strains of T. vaginalis examined were sensitive to metronidazole (MDE: less than 3 x 35 mg/kg peroral).


Secnidazole. A 5-nitroimidazole derivative with a long half-life. The therapeutic activity of a single 2 g dose of secnidazole was studied in patients with urogenital trichomoniasis. In 140 patients, 97% were cured and the drug was well tolerated. In the laboratory, tests on sensitivity were made and the minimal inhibitory concentration (MIC) and the minimal trichomonacidal concentration (MTC) were determined on cultures that had recently been isolated at the clinic, and the pharmacokinetic properties of secnidazole in man were compared with those of tinidazole. The therapeutic efficacy of all the metronidazole derivatives was reviewed and a single-dose treatment is recommended. Therapeutic and prophylactic treatment is achieved by products with a long half-life. Secnidazole, with a half-life of 14.3 +/- 1.3 h (women) and 20.2 +/- 3.1 h (men), is particularly suitable for this type of treatment.


Metronidazole and tinidazole in a single large dose for treating urogenital infections with Trichomonas vaginalis in men. Seventy-three men with urogenital trichomonal infection were treated with a single daily dose of 1 g tinidazole or 1.5 g metronidazole. Both treatments gave satisfactory results. Wives who were also infected with Trichomonas vaginalis, were less likely to have a recurrence of the infection after their husbands had been given either drug.


Abnormal forms of Trichomonas vaginalis. Abnormal forms of Trichomonas vaginalis have been demonstrated by both conventional and scanning electron microscopy after inoculation of media with clinical material from cases of trichomonal vaginitis. Twenty-six cases of vaginitis have been studied; 10 of them showed the abnormal forms of trichomonads after growth in a modification of the medium described by Bushby and Copp (1955), while 16 showed only normal forms.


GABA-mediated control of transient signals in the inner retina. The effects of chemicals on visual activity was investigated by recording the proximal negative response (PNR) in the perfused eyecup of frogs. Various amino acids reversibly decrease or abolish the PNR over an equivalent concentration range but differ in this depressive action in the presence of convulsant alkaloids. Depression produced by GABA was antagonized by picrotoxin or bicuculline; while glycine-produced depression was selectively antagonized by strychnine. In addition to selective antagonism of amino acids, the convulsants produced differential enhancement of sustained and transient phases of the PNR. In this respect, the effects of picrotoxin and bicuculline are similar to those of semicarbazide and chloride-free solutions which may also inactivate GABA pathways. The effects of these chemicals on the time course and input-output relation of the PNR suggest that processes involving GABA may regulate the transformation from sustained to transient signals which occurs in the inner retina.


The course of direct projections from the abducens nucleus to the contralateral medial rectus subdivision of the oculomotor nucleus in the cat. We have used autoradiography (tritiated leucine) to investigate the projections of a number of nuclear groups of the cat pons. Some cells of the abducens nucleus have axons that cross the midline, ascend in the opposite median longitudinal fasciculus (MLF) and synapse on the cells of the oculomotor complex which have been identified by others as those innervating the medial rectus muscle.


Congenital nystagmus--genetic and environmental causes. A survey of 40 individuals registered with the Canadian National Institute for the Blind (CNIB) as blind from congenital nystagmus revealed that an abnormal single gene was responsible for the disorder in 33 patients. Fifteen of these were due to autosomal recessive conditions while X-linked disorders accounted for another 15 patients. In 3 cases the pedigrees were consistent with both autosomal recessive or X-linked inheritance. A clearly defined environmental origin was present in 1 case while specific genetic or environmental factors were not detected in the remaining six patients. The albinism, achromatopsia and Leber's congenital amaurosis groups of disorders were those most frequently detected.


The effects of acute hypercapnia on cerebral indole amine metabolism. The effects of 1-h exposure to hypercapnia (PaCO2, 90-110 MMHg) on cerebral indole amine metabolism were studied in rats by measurement of cerebral hemisphere contents of tryptophan, 5-hydroxytryptophan (5-HTP), 5-hydroxytryptamine (5-HT), and 5-hydroxyindoleacetic acid (5-HIAA), 5-HIAA content was increased after 1-h exposure to hypercapnia, whereas tryptophan, 5-HTP, and 5-HT remained unchanged from control. The accumulation of 5-HTP after decarboxylase inhibition with 3-hydroxybenzyl hydrazine was increased in hypercapnic rats and indicated an increased activity of tryptophan hydroxylase. During the 1-h exposure to hypercapnia there was increased accumulation of 5-HT after monoamine oxidase inhibition with pargyline and increased accumulation of 5-HIAA arter probenecid. The results indicate an increased synthesis and degradation of indole amines in acute hypercapnia.


Rheumatoid factor positive plasma and humoral cytotoxicity to malignant melanoma. Complement dependent cytotoxicity to malignant melanoma tumor cells was demonstrated in incubations that included rheumatoid factor (RF) positive plasma and normal plasma. Cytotoxicity was not demonstrated to cells from a number of normal tissues. The phenomenon involved coating of tumor cells with immunoglobulins and complement fixation. The activity in RF positive plasma was present in high titer an may not be RF. The activity in normal plasma was present in low titer, was found in plasma from eight out of 11  healthy subjects and may be a naturally occurring antibody. This previously undescribed humoral cytotoxicity system may participate in tumor host interactions, especially after therapy, when the majority of patients become RF seropositive.


Inhibition of the phosphorylation of non-histone chromosomal proteins of rat liver by cordycepin and cordycepin triphosphate. The effects of cordycepin (3'-deoxyadenosine) and cordycepin triphosphate on phosphorylation of non-histone chromosomal proteins were assessed in isolated hepatic nuclei in vitro. Cordycepin and cordycepin triphosphate competitively inhibited phosphorylation of urea-soluble nuclear proteins with a Ki of 1.2 X 10(-3) and 8 X 10(-5) M, respectively. Isoelectric focusing of urea-soluble proteins indicated that inhibition occurred predominantly in nuclear proteins with isoelectric points of pH 4 to 7. Quaternary aminoethyl-Sephadex chromatography of extracts of nuclei incubated with cordycepin and cordycepin triphosphate also showed inhibition of phosphorylation of non-histone chromosomal proteins with similar isoelectric points, although greater resolution of proteins with isoelectric points of pH 6 to 7 was achieved. RNA polymerase I and II were not affected by cordycepin and cordycepin triphosphate after quaternary aminoethyl-Sephadex chromatography of nuclear extracts incubated with either agent. However, RNA polymerase I and II in isolated nuclei were competitively inhibited by cordycepin triphosphate but not by cordycepin. These results suggest that cordycepin triphosphate, and perhaps cordycepin too, may affect transcription via interference with the phosphorylation of non-histone chromosomal proteins.


Induction of myeloid colony-stimulating activity in murine monocyte tumor cell lines by macrophage activators and in a T-cell line by concanavalin A. Certain fibrosarcoma lines in culture and the WEHI-3 myelomonocytic leukemia cell line have previously been shown to secrete myeloid colony-stimulating activity (CSA) spontaneously. We describe here other hematopoietic tumor cell lines in which CSA is either produced constitutively or inducible by immunostimulators. CSA production in macrophage and monocyte tumor lines is induced by lipopolysaccharide, zymosan, Mycobacterium strain Bacillus Calmette-Guérin, tuberculin purified protein-derivative preparation from mycobacteria, and dextran sulfate. Myeloma, mastocytoma, and T-lymphoma lines do not produce CSA with or without these agents. In contrast, the T-lymphocyte mitogen concanavalin A (but not phytohemagglutinin) induces CSA synthesis in one of seven T-lymphomas tested. In most cases induction of CSA is correlated with conditions of cell growth inhibition by the immunomodulators. However, other drugs that cause cytostasis or cytotoxicity do not lead to CSA production. Leukemic cells thus may retain sensitivity to normal regulatory events with resultant effects on host hematopoietic cell functions.


Electron microscopic localization of 3beta-hydroxysteroid dehydrogenase and NADH-ferricyanide reductase activities in amphibian interrenal cells. 3beta-hydroxysteroid dehydrogenase and NADH-ferricyanide reductase activities were localized at the ultrastructural level in amphibian interrenal (adrenocortical) cells previously fixed in a mixture of formaldehyde and glutaraldehyde. Potassium ferricyanide was used as an electron acceptor. Copper ferrocyanide deposits resulting from 3beta-HSD activity were seen in close association with the external faces of the membranes of the smooth endoplasmic reticulum. Very rare grains of precipitate appeared in mitochondrial cristae. The addition of phenazine methosulfate to the incubation medium had no effect on these localizations. The interrenal cells showed also a strong NADH-ferricyanide reductase activity. The copper ferrocyanide grains were abundant in the mitochondrial cristae and in the hyaloplasm, where they were not preferentially associated with the smooth endoplasmic reticulum.


Left main coronary artery stenosis: the influence of aortocoronary bypass surgery on survival. A retrospective study was carried out in 114 unoperated and 197 operated patients having left main coronary artery (LMCA) stenosis greater than or equal to 50%. Including the operative mortality of 9.1%, survival at seven years was significantly greater following pure aortocoronary bypass graft surgery, 77.5% as compared to 48.5% for the unoperated patients (P less than 0.01). The surgical mortality was significantly less during the last five years (1972-1976), 6.2% as compared to 17% during 1969-1971 (P less than 0.025). The three year survival in patients operated since 1972 was 90.2% as compared to 60.4% for unoperated patients. Survival remained significantly higher in the operated patients when studied as subsets on the basis of the severity of the LMCA stenosis (less than 70% as opposed to greater than or equal to 70%), and on the extent of associated obstructive disease of major coronary arteries (0-1 versus 2-3 arteries). It was significantly higher, however, only in operated patients with associated stenosis greater than or equal to 70% of the right coronary artery. Survival was higher following surgery only when the ejection fraction was at least 0.45, or the left ventricular end-diastolic pressure below 20 mm Hg.


Albumin activation of urinary amylase as determined with the Du Pont aca. Protein activation of urinary alpha-amylase (EC 3.2.1.1) activity was observed during an evaluation of the Du Pont aca procedure for the determination of urinary alpha-amylase. This activation effect became constant for urinary albumin concentrations exceeding 1.50 g/liter. It is recommended that urinary alpha-amylase be analyzed with sufficient albumin added to maximize this effect. The aca alpha-amylase procedure is compared to an amyloclastic method for both serum and urine analysis. Expected ranges are presented for the aca method for serum and urinary amylase, amylase clearance, and the amylase clearance/creatinine clearance ratio.


Acyl derivatives of p-aminobenzoic acid as new substrates for the assay of serum acylase activity. A simple colorimetric method for the assay of cobalt-activated acylase activity in human serum using new and less toxic N-chloroacetyl-gamma-L-glutamyl-p-aminobenzoic acid as substrate has been described. The values obtained with this method are almost the same as with the previously described method using naphthylamide substrate.


Distribution of mononuclear cells during pregnancy. Leucocyte and lymphocyte counts, the distribution of erythrocyte rosette-forming cells (E-RFC), erythrocyte-antibody-complement rosette-forming cells (EAC-RFC), cells bearing surface membrane immunoglobulin (SMIg), latex-ingesting cells and the PHA stimulation index of lymphocytes were evaluated in both pregnant and non-pregnant women. In the gravid female leucocytosis due to neutrophilia is seen, and the PHA-induced transformation of maternal lymphocytes in the absence of autologous plasma remains unaltered. There was a rise in the percentage of EAC-RFC and cells bearing SMIg during a pregnancy, but after monocyte adsorption on plastic dishes of the mononuclear cells, obtained after Ficoll-Hypapue centrifugation, this increase is abolished. There is a significant increase of the number of latex-ingesting cells during pregnancy. These findings indicate a significant increase of the monocytes during pregnancy.


[Age and immunity. I. Lymphocyte populations in the peripheral blood in various age groups (author's transl)]. The total lymphocyte count and the different lymphocyte subpopulations were determined in peripheral blood of healthy probands aged 20 to 90 years. With increasing age there is a slow decrease of the total lymphocyte count and of the subpopulations. This was most evident in the decrease in thymus dependent lymphocytes (T cells) which carry cellular immunity. These results support the assumption that with increasing age cellular immunity is clearly reduced and may possibly be associated with an increased incidence of neoplasia, infection, and auto-immune disease. Estimations of lymphocytes and their subpopulations in peripheral blood allow some conclusions as to the immunological resistance which is largely dependent on these cells. Estimation of the number of circulating lymphocytes in relation to age is thus indispensable for the assessment of the immunological reactivity of patients.


[Infectious resistance to antibiotics in Haemophilus influenzae (author's transl)]. Ampicillin-resistant Haemophilus influenzae does occur now in the FRG. In one isolate a plasmid with resistance genes (R-factor) could be demonstrated as cause of the ampicillin resistance. This R-factor influences production of a beta-lactamase of the TEM type which destroys ampicillin. The infectious nature of the ampicillin resistance was proven by the fact that it was transferable to other bacterial species through cocultivation. Parallel to ampicillin resistance tetracycline resistant Haemophilus influenzae has occurred in the FRG. Here the resistance was equally bound to plasmids. These R-factors are infectious as well. Molecular analysis of the 3 isolated resistance factors in Haemophilus influenzae showed that they carry the same resistance genes which are known from R-factors of Enterobacteriaceae. In the therapy of purulent infections due to Haemophilus influenzae such as childhood meningitis one can no longer rely on general ampicillin sensitivity of the offender. Apart from ampicillin and tetracycline resistant Haemophilus influenzae chloramphenicol resistance has been observed in a few cases.


The use of low intensity electrical stimulation to the auricles in psychiatric patients. The regulated passage of an intense convulsion producing electrical current through the brain as in Electro Convulsive Therapy (ECT) is an accepted modality of psychiatric therapy. The purpose of this study was to examine the effects of a brief cyclic electrical current to both auricles. The current was tolerable, nonconvulsive in nature and self-administered by the patients at home. The data indicates the procedure proved a very potent method for the control of mental depression, anxiety-depression and delusional states. In the majority of cases, it completely eliminated, for many months after treatment, the need for psychiatric or drug supportive therapy.


The immunological consequences of nematode infection. Nematode infections in the gut induce a strong immune response which is rapidly detected parenterally. The response is thymus-dependent and long-lasting and involves both antibodies and cell-mediated reactions. The immunological response to unrelated antigens, tumours and other infectious organisms is altered in animals infected with nematodes. Both antibodies and sensitized lymphocytes participate in the immune response which affects the nematodes themselves, and characteristically the lymphocyte-dependent step cannot act in lactating animals and is neither induced nor able to act in young animals. Present evidence suggests that, despite their well-known association with helminth infections, parasite rejection from the gut does not require the participation of IgE antibodies, mast cells or eosinophils. Homing of lymphoblasts from the mesenteric lymph node or thoracic duct lymph to the small intestine is increased in rats and mice infected with Nippostrongylus brasiliensis or Trichinella spiralis and the increase is antigenically non-specific. In mice infected with T. spiralis this increase is represented mainly by thymus-derived lymphoblasts.


Effect of neonatal corticoid treatment on tryptophan and serotonin metabolism. Glucocorticoid administration to newborn rats leads to the premature induction of hepatic tryptophan oxygenase. This is accompanied by a retardation of the developmental increase in brain serotinin both in regions of serotoninergic cell bodies and in terminal projection areas. The magnitude of the fall in serotonin is relatively small. This appears due, at early ages to the concomitant induction of brain tryptophan hydroxylase and at late periods to retarded development of the blood brain barrier against tryptophan.


Effects of phentolamine and clonidine on pituitary-adrenocortical axis during stress in conscious rats. For evaluation of the effect of several drugs on the hypothalamo-pituitary-adrenocortical axis during stress, after testing various tress stimuli (swimming, restrain) the 15 min exposure of rats to novel environment was chosen. Subcutaneous administration of clonidine hydrochloride (BOEHRINGER; Ingelheim, 60 micorgram kg-1), a drug stimulating alpha-adrenergic receptors, did not result in any any changes in plasma corticosterone level either during basal or under stress conditions. In contrast, the administration of alpha-adrenergic blocking agent--phentolamine (RegitineR CIBA; 4 mg kg-1)--produced a significant elevation of resting corticosterone level, but failed to influence the stress-induced corticosterone increase.


Binding of 3H-dexamethasone and 3H-corticosterone to rat liver cytosol during postnatal period. The binding of 3H-dexamethasone (3H-Dx) and 3H-corticosterone (3H-B) to specific binding proteins of rat liver cytosol was compared during pre- and postnatal period. The binding of 3H-Dx was almost twice as high as that of 3H-B during the whole postnatal period. The Scatchard analysis showed a number of binding sites for 3H-Dx to be 4.44 X 10(-13) mol mg-1 (Kd 5.9 X 10(-9) mol 1(-1)) and for 3H-B to be 2.7 X 10(-13) mol mg-1 (Kd 1.6 X 10(-8) mol 1(-1)). A negative relationship between the binding of 3H-Dx to the rat liver cytosol receptors and the level of plasma corticosterone was found and it was concluded that thenumber of free receptors is, to the certain extent, dependent on thelevel of endogenous corticosteron. In spite of that, however, it seems that, in addition to corticosterone, the undetectability of receptors during a short period after birth may be due to some other factors.


Influence of insulin on mitotic rate in cultivated mammalian cells. The influence of insulin (100 mU ml-1) on the mitotic rate in a tissue culture of muscle fibroblasts L was studied. The metod of temporary inhibition of cell transformation from G1 to S period with the aid of maintaining cells in serum free medium was used. Moreover, the number of mitoses in a stage of metaphasis after the addition of colchicine was counted and autoradiography was used. Insulin showed two independent effects on mitotic cycle: first it initiated the synthesis of DNA and increased the rate of a transformation of cells from a period G1 into period S; second--it increased the rate of processes at the final stages of mitotic cycle and increased the number of cells entering this cycle.


Effect of hydrocortisone on bioelectric activity and morphological pattern of spinal cord in frogs. The effect of 1 mg hydrocortisone (HC) administered 3 h before the experiment was studied the bioelectric activity of ventral roots of spinal cord, the size and morphology of motoric neurons and also the size of synaptic endings being measured under different frequency of electric stimulation in frogs. It was found that the effect of HC on the electric activity of spinal cord and morphology of its neural elements was dependent on the stimulation used. Under the resting conditins the administration of HC resulted in a small decrease of the size of neurons. Under a low frequency stimulation HC did not affect the electric activity of spinal cord, but prevented the swelling of neurons and of synaptic endings resulting from the stimulation. Under a high frequency stimulation themorphology of neurons was remarkably changed, a shrinkage of these and an appearance of "dark" cells being found. Also these changes were prevented by the administration of HC. It was concluded that in all experiments HC prevented the changes resulting from electrostimulation, its effect, however, being dependent on the parameters of the stimulation used.


Lesions of some central nervous structures result in altered pituitary-thyroid response to acute decrease of blood thyroid hormone level. Eight groups (3--6 animals each) of rats weighing 350--400 g were subjected to electrolytic lesions of various parts of habenular, thalamic and hypothalamic areas of brain. On the 6th day after the lesion the level of blood thyroid hormone was acutely decreased with the aid of isovolemic exchange transfusion (IET) of thyroid hormone free blood suspension. The level of thyroxine (T4) in plasma was measured before and during 180 min after IET with the aid of specific radioimmunoassay and the changes of its post-transfusion level were evaluated. It was found that in three groups of animals bearing large bilateral lesions either in lateral ventral thalamus or in a central inferior thalamus the response of T4 level during the post-transfusion period is similar to that found in intact control groups from previous experiments. This consists in an increase of T4 level nearly to the initial value within about 120--150 min after IET. In contrast, there was only a slight post-transfusion increase of T4 level in one group with small central lesion in medial superior thalamus and no increase in two groups bilaterally lesioned in habenular area and in another two groups lesioned either in a central part of dorsal hypothalamus or in a central dorsal part of ventral basal hypothalamus. It was concluded that some parts of brain may be involved in managing the appropriate response of pituitary-thyroid axis to acute decrease of thyroid hormone level, no plausible explanation of the mechanism ofthe observed phenomena being offered.


Activation of human B and T lymphocytes by protein A of Staphylococcus aureus. Protein A from Staphylococcus aureus, in soluble form or coupled to Sepharose beads, acts as a polyclonal B cell activator (PBA) for human lymphocytes in blood and spleen. PBA activity was demonstrated in spleen cells by the ability of protein A to induce the formation of intracellular immunoglobulin synthesis and to activate polyclonal antibody secretion demonstrated against fluorescein isothiocyanate-coupled sheep erythrocytes in a modified hemolysis in gel assay. More plaqueforming cells (PFC) were seen in unseparated cells than in purified B cells. In blood lymphocytes, only few PFC were activated by soluble protein A. Protein A increased DNA synthesis in blood and spleen cells. At a concentration of 100 microgram/ml the peak response was on day 4 or 5, but at 1 microgram/ml the peak response occurred later. On day 4 of culture, high mitogenic activity was seen in unseparated lymphocytes or mixtures of separated B and T cells, whereas in enriched B and T cell suspensions activity was low. On day 7, however, DNA synthesis in both the enriched B and T cells was higher than in mixtures of B and T cells. Protein A stimulated DNA synthesis in thymus cells with a peak response on day 6. It is concluded that protein A alone or as an IgG complex can activate both B and T cells, though the mechanism of activation is not known and may be different for B and T cells.


Genetic and functional characterization of an antiserum to the lipid A-specific triggering receptor on murine B lymphocytes. The inheritance of responsiveness to lipopolysaccharide (LPS), and of a marker recognized in LPS-reactive cells by a heterologous antiserum, was studied in crosses between C3H/HeJ (nonresponder) and C3H/Tif (high responder) mice. F1 hybrid mice show codominant expression of these traits: (a) LPS-reactive cells are only hlaf as frequent in the hybrids as in the high responder parent; (b) the serologically defined marker is expressed in half as many cells in the hybrids as in the high responder parent. In backcross generations, both LPS responsiveness and this serological marker segregated into high, intermediate, and nonresponders. LPS or free lipid A, but not two other B cell mitogens (lipoprotein, and purified protein derivative of tuberculin), compete with the antiserum for binding to the B cell surface membrane, and are capable of completely inhibiting such binding without interfering with the binding of a-ti-Ig antibodies or complexes to Fc receptors. The addition of an IgG fraction of the antiserum to B cell cultures results in exponetial growth of the cells and in maturation to antibody secretion. This mitogenic activity is dose-depedent and absorbable on spleen cells from LPS high responder mice. Taken together, these observations suggest that this antiserum contains antibodies to the lipid A-specific triggering receptor on B lymphocytes.


Cell membrane-mediated cytolysis by membranes from noncytolytic cells. Cytolysis was obtained with plasma membrane fractions prepared not only from cytolytic T cells, but also from nonimmune lymphoid cells and, moreover, from cell lines of nonlymphoid origin displaying no lytic ability. Furthermore, no relationship exists between the genetic specificty of T cell-mediated cytolysis and the activity of membrane preparations. If anything, it appears that certain cell types may be unusually sensitive to membrane-mediated cytolysis. Our observations argue against a mechanistic relationship between T cell-mediated and membrane-mediated cytolysis.


The effects of anti-theta-globulin treatment on the morphology of the mouse spleen. Rabbit anti-mouse-theta-globulin raised against the brain tissue of DBA/2 mice was injected into DBA/2 mice subcutaneously 4 times at 2 days intervals and the histology of the spleen was assessed 1, 2, 4, 7, 14 and 21 days after the last dose. Untreated animals and those treated with normal rabbit gammaglobulin were used as controls. Anti-theta-globulin was found to induce a profound depletion of the small lymphocytes in the central regions of the periarterial lymphoid sheath (C-PALS). At the same time, a strong stimulation of the cellular elements involved in antibody response was evident in the peripheral regions of the periarterial lymphoid sheath (P-PALS). This stimulation was even more pronounced in mice treated with normal rabbit gammaglobulin, a fact possibly due to the undisturbed T-B-cell interaction in these cases. The results support the concept that two distinct areas exist in the spleen white pulp, C-PALS and P-PALS, a thymus- and a bursa-dependent area, respectively. A standardized system, analogous to that previously proposed for the morphology of the lymph nodes by Cottier et al. (1973), was introduced to describe the morphology of the spleen white pulp in relation to the immunological function.


Effects of the antiprotease Trasylol on peripheral blood leucocytes. Binding of the antiprotease Trasylol to human peripheral blood lymphocytes and polymorphonuclear leucocytes (PMNs) was demonstrated at the ultrastructural level using an indirect immunoperoxidase technique. This also revealed endocytosis of membrane bound Trasylol by PMNs. Trasylol inhibited PHA- and ConA-induced lymphocyte stimulation, and was cytotoxic to unstimulated cells.


Kinetic analysis of the enhanced immune responsiveness of baby chicks grafted with lymphoid cells from juvenile donors. The immunologic immaturity of embryos and newly hatched chicks to MRBC immunization has provided us with a model to investigate the distribution of immunocytes and their precursors in several tissues of juvenile chickens. Antigen-responsive cells (ARC) were rare in the peripheral blood, spleen, bone marrow, thymus and bursa of Fabricius of unimmunized young birds. Following MRBC immunization, they were abundant in the blood and spleen, low in frequency in bone marrow and absent in the thymus and bursa. The recipients of bone marrow cells, thymocytes, or bursacytes, when immunized at hatching, exhibited accelerated humoral immune responses. The enhancement property of the grafted cells was increased when MRBC was included in the cell suspension at the time of grafting. The precocious responsiveness of the grafted chicks is attributed to the cooperation of donor and host immunocytes. The enhancement effect, moreover, was less consistently expressed in allogeneic hosts. This capacity was aquired by thymic cells within a day after immunization of the donor and persisted well over a week. The role of antigen in driving the specific differentiation of T and Bu cells is briefly discussed.


Serum alpha 1-antitrypsin levels in alcoholic hepatitis. Serum alpha 1-antitrypsin concentrations have been measured in 68 patients with liver disease. High mean values (359 +/- 18.0) were found in patients with alcoholic hepatitis whereas patients with acute viral hepatitis and chronic active hepatitis did not show any significant differences from the controls. In a group of patients with both alcoholic hepatitis and acute pancreatitis the mean value (218 +/- 5.8) was significantly lower than in the control group. The mechanism for the reduction of serum alpha 1-antitrypsin levels in this group remains to be clarified but low serum alpha 1-antitrypsin may be due to increased proteolytic enzyme concentrations in acute pancreatitis.


Establishment of a T-cell line from human lymphosarcoma. A leukemic T-cell line, designated as TALL-1, has been established in continuous culture from the bone marrow of a patient with a leukemic phase of T-cell lymphosarcoma. TALL-1 cells have T-cell properties and lack Epstein-Barr virus or its genome. The cells of TALL-1 line are considered to have originated from the donor's leukemic cells on the basis of their cytogenetic, morphological, and surface features.


Eosinophilic gastroenteritis with obstruction. Immunological studies of seven patients. We studied 7 patients with eosinophilic gastroenteritis presenting with obstruction of the gastrointestinal tract. Five had required surgical treatment for relief of obstructive symptoms or because of diagnostic uncertainty. Immunological studies, including serum immunoglobulins, serum complement, lymphocyte quantitation, and lymphocyte response to nonspecific mitogens revealed no striking abnormalities. None was clinically allergic or food sensitive, and serum IgE was normal in all. A review of the literature since 1970 suggests that the obstructive form of eosinophilic gastroenteritis accounts for about half of the reported cases. Allergy, especially hypersensitivity to food, is not required for the diagnosis.


[Animal experiments on the prevention of peritoneal adhesions (author's transl)]. Several possibilities for the prevention of adhesions were checked in animal experiments. In rats adhesions were produced by a standardized operative procedure. For the prevention of adhesions an artificial ascites with normal saline solution or Dextran was tried. To maintain ascites an intraperitoneal catheter was left in place for the first three post-operative days and refilled. The proteolytic effect of Chymotrypsin, the proteinase inhibitor Trasylol and of Glucocorticoids was investigated. All three drugs showed a definite therapeutic effect. In 45-50% of the rats adhesions were prevented. A combination of Trasylol, Glucocorticoids and Dextran prevented adhesions in 65% of the animals. When the ascites was maintained for four days 80% of the animals remained free of adhesions.


[Interactions between 6-chloro-5-cyclohexyl-1-indancarboxylic acid (TAI-284) and biopolymers (author's transl)]. TAI-284, a new non-steroidal acidic analgesic and anti-inflammatory agent was investigated and the interactions with biopolymers were compared with those of indomethacin (IMC) and ibuprofen (IP). TAI-284 inhibited the heat denaturation of bovine serum albumin at pH 5.3, similar to that seen with IMC and weaker than that seen with phenylbutazone. TAI-284 prevented the rat erythrocyte from heat-induced hemolysis and was twice as potent as IMC. TAI-284 produced alterations in platelet function as characterized by loss of secondary aggregation in response to ADP and inhibition of aggregation by collagen. Both these effects were one fifth as potent as those seen with IMC. In rats, platelet aggregation induced by collagen and secondary ADP aggregation was reduced in a dose-dependent manner by a single oral administration of TAI-284. The inhibitory activity was approximately one fourth that of IMC or twice that of IP in the former and in the latter one fifth that of IMC or similar as that of IP. These results suggest that the essential feature of TAI-284 is its potent membrane stabilizing action which is considered to be an necessary mechanism in the action of anti-inflammatory drugs. It is considered that TAI-284 may be less active than IMC in inhibiting prostaglandin biosynthesis in platelets.


[Angiographic diagnosis and therapy of acute and chronic gastrointestinal hemorrhages]. In acute gastrointestinal bleeding visceral angiography has been showing its importance for years. It contributes to diagnosis especially in cases with persistent acute hemorrhage. In chronic gastrointestinal bleeding conventional radiographic procedures such as upper gastrointestinal series and barium enema will be preferred to angiography. The function of the radiologist goes beyond mere diagnosis of gastrointestinal bleeding. Treatment with vasopressin via the angiographic catheter has proven its clinical value. This method will be indicated especially in cases with high risk anesthesia and surgery. It will help to postpone necessary surgery to a more favorable moment following hemostasis. Side effects such as hypertension and antidiuresis are relatively rare and easy to manage. Numerous substances are used for embolization showing that ideal material has not been found yet and further development seems necessary. In contrast to vasopressin treatment, vascular occlusion is often irreversible, complications (unwanted reflux of embolization material, necrosis and plugging of the catheter) are more difficult to manage. Superselective visualization of a bleeding artery is always needed. Embolization is justified in cases when a possibility for anesthesia and surgery cannot be foreseen. The electrical vascular occlusion using direct current is still in the phase of animal experiments; its clinical value has not sufficiently been assessed as yet.


Delayed hypersensitivity in mice induced by intravenous sensitization with sheep erythrocytes: evidence for tuberculin type delayed hypersensitivity of the reaction. Delayed hypersensitivity (DH) reaction can be induced in mice by intravenous sensitization with sheep erythrocytes (SRBC). However, as the sensitizing procedure is quite different from a usual mode of sensitization for DH using complete Freund's adjuvant (FCA), the nature of this reaction has been a matter of controversy. In an attempt to characterize this reaction, we placed special interest on two possibilities regarding the nature of this reaction; Jones-Mote reaction or tuberculin type DH. From the kinetics study on the DH after challenge, the DH reaction to SRBC in mice by intravenous sensitization was clearly distinguished from the Arthus reaction. The dose-response pattern of this reaction also suggested that the contribution of Arthus reactivity to delayed reactivity was negligible. Cell reconstitution experiments revealed this DH to be quantitatively thymus cell dependent. Furthermore, this DH required macrophages at its manifestation stage, and appearance of basophil infiltration at the lesion was absent. In addition, strain difference and ageing of host mice influenced the DH reaction in exactly the same fashion in which these factors influence the tuberculin type-DH induced by subcutaneous sensitization with methylated human serum albumin (MHSA) in FCA. Taken collectively, it was concluded that this DH reaction can be categorized as the tuberculin type.


Antibody responses to contact sensitizing agents. Effect of sensitized cells. Lymphocytes from mice immunized by painting with the contact sensitizing agent picryl chloride have been shown to produce regulatory effects on the DNA synthesis and contact sensitivity responses of normal mice painted with picryl chloride. This report describes the effect of these cells on antibody responses of normal mice to picryl chloride. Lymph node cells taken 5--7 days after painting increased early IgM antibody responses of normal mice to picryl chloride. Spleen cells were not effective. The increase was mediated by T cells as judged by anti-theta treatment and nylon wool filtration and could not be produced by killed, irradiated or allogeneic cells. A similar activity could be demonstrated in cells from mice painted with another contact sensitizing agent, oxazolone. The effect was strictly specific and cells from mice painted with picryl chloride or oxazolone would only increase responses to picryl chloride or oxazolone, respectively, even when presented with a mixture of the two antigens. The cel-s increasing antibody production were different from T cells previously shown to mediate contact sensitivity and T cells shown to regulate DNA synthesis in that they could not be generated in adult-thymectomized mice.


Active suppression of in vitro reactivity of spleen cells after BCG treatment. It was found that spleen cells from mice injected i.v. with large doses of BCG respond to PHA stimulation less intensely than do normal spleen cells. We were able to show that nylon wool column purified BCG treated T cells also had a low PHA reactivity. Unfractionated spleen cells, adherent cells or T-enriched populations from BCG treated mice, when added to normal T cells lowered their PHA reactivity. When the same BCG treated cell populations were added to tumor cells in vitro, they inhibited their growth.


Inhibition of the classical and alternative pathways by amino acids and their derivatives. Effects of various aminoacids and their derivatives on the classical pathway and alternative pathway of the complement were studied. Leupeptin, acetyl-leucyl-leucyl-arginal, inhibited CH50 and Cl-esterase, but did not inhibit the alternative pathway. When aminoacids of carbon chains of the order of seven were used, arginine and lysine had stronger effects than trans-aminomethyl cyclohexane carboxylic acid (t-AMCHA), cis-aminomethyl cyclohexane carboxylic acid (cis-AMCHA) and epsilon aminocaproic acid (EACA). SH-compounds, cysteine, homocysteine and glutathione, had the strongest inhibitory effects among these aminoacids on both classical and alternative pathways. When effects on Cl esterase were compared, arginine, lysine, t-AMCHA, cis-AMCHA and EACA had weak inhibition while SH-compounds showed strong inhibition. Poly-L-lysine, which had extremely strong inhibition of CH50, had no inhibition of Cl esterase. The inhibitory effects of antifibrinolytic agents, EACA and t-AMCHA, were weak but when effects on early parts of the classical pathway, C(1,4,2)H50 were tested, some inhibitory activities were recognized. Thus inhibitory effects of these agents were due to their activities on the early parts of the classical pathway.


Relationships among differentiated T-cell subpopulations. III. Radioresistance of delayed hypersensitivity to heterologous erythrocytes. Effects of whole body irradiation with 600 rad on delayed hypersensitivity, direct cytotoxicity and antibody production were examined in mice immunized with chicken erythrocytes (CRBC) or sheep erythrocytes (SRBC) in saline. Delayed footpad reactions to CRBC were not affected by the exposure to irradiation 3 h before or after antigenic stimulation. On the other hand, direct cytotoxic activities in spleen cells and antibody titres to CRBC were reduced by such exposures. Additionally, delayed footpad reactions to SRBC were not affected by the exposure to irradiation. Generation of effector cells responsible for delayed footpad reactions proved to be resistant to irradiation.


Partial purification and characterization of the human lymphocyte colony enhancing factor (LCEF). The cloning potential of PHA-treated T cells is significantly enhanced when lymphocyte culture fluid (LCF) from mitogen-treated lymphocytes is added to the soft agar culture system. The mitogens seem to stimulate the release of a lymphocyte colony enhancing factor (LCEF) into the culture medium. A study of the physico-chemical properties of the LCEF revealed that it is a nondialyzable, heat-labile molecule which migrates in the haptoglobin (2--2) post-transferrin region in acrylamide electrophoresis. It is stable to RNase and DNase but labile to papain and trypsin. The LCEF was partially purified from the crude LCF using a sequence of techniques--ammonium sulphate precipitation, DEAE-cellulose and Bio-Gel P-150 chromatography and disc electrophoresis. The mol. wt of the purified LCEF, determined from gel filtration chromatography, was 90,000--110,000.


Differences in lifespan and rate of turnover between phytohaemagglutinin responsive cells of the bone marrow and of peripheral lymphoid organs. Radioautographic analyses were performed on PHA stimulated cultures of in vivo labelled cells obtained from mice previously injected with 3H-TdR to selectively label either cells with a rapid renewal rate (RR) or a slow renewal rate (SR). ha responsive cells in the bone marrow (BM) were found to be virtually all RR cells, whereas both RR and SR cells from lymph nodes (LN) and spleen (Spl) were stimulated by this mitogen. However, RR cells were proportionately more responsive to PHA than SR cells in all tissues examined. Only one out of 200 BM PHA blasts belonged to the SR subclass, whereas the RR/SR ratio was approximately 1/1 for LN and 2/1 for Spl. Control experiments demonstrated that significant in vitro re-utilization of 3H-TdR from dying cells did not occur in the cultures. These results support a growing body of evidence that BM PHA responsive cells are precursor T-cells which are known to have a rapid turnover rate.


Resistance and susceptibility of mice to bacterial infection: genetics of listeriosis. A survey of various strains of mice showed distinct differences in resistance or susceptibility to Listeria monocytogenes. C57B1, related sublines, NZB, and SJL were resistant to Listeria, whereas BALB/c, CBA, A, DBA/1, C3H, LP.RIII, 129, and WB were susceptible. The gene(s) responsible for resistance and susceptibility to Listeria were studied in detail. C57BL6/6, B10.D2, and B10.A mice were 100 times more resistant than were BALB/c, CBA, and A. Resistance of the (C57B1/6 X BALB/C)F1 was intermediate between the two parents, suggesting partial penetration of a dominant gene. Backcross studies in which the (C57B1/6 X BALB/c)F1 were crossed with the susceptible BALB/c parent suggested that a single gene or group of linked genes were the major determinant of resistance, although the possibility that other genes exerted a modifying influence was not excluded. By using the backcross and various congenic and recombinant mice, linkage of the genes involved to the H-1, H-2, H-3, H-4, H-7, or H-8 loci, to the immunoglobulin allotype, to the Thy-1 gene, to the Hc gene specifying C5, or to coat color genes (B, c) was excluded. There was no difference in the response of males and females. In all studies, the powerful overriding influence of the C57B1 genome was evident.


New methods of pain measurement and their application to pain control. Recent pain research advances show promise in their application to the relief of acute and chronic clinical dental pain. Regional electroanalgesia, or transcutaneous electrical stimulation, has been used successfully in the treatment of pain associated with peripheral nerve injuries. Electrical stimulation of teeth also may prove useful as a pain control technique during operative dentistry procedures. Another exciting research finding is the discovery of endogenous or natural pain-suppressing pathways in the brain. There are recent demonstrations that natural-occurring opiate-like compounds and receptors exist in the brain. The elucidation of stimuli and behavioral responses which will activate these specific descending pain control pathways may lead to exciting new methods of pain relief. Thus, both regional electroanalgesia and the discovery of endogenous pain-suppressing pathways offer the possibility of the future expanded use of non-pharmacological pain control techniques. The proper evaluation of new pain control techniques requires the development of better methods of measuring and assessing the multidimensional aspects of the pain experience. Category scales which scale the suprathreshold range of pain from threshold to tolerance levels can be used with both experimental and clinical pain. Sensory Decision Theory has been applied to the analysis of categorical pain responses. This method distinguishes between sensitivity to stimulus intensity and response bias, or the patient's willingness to report a given intensity as painful. Another promising method for scaling pain is the use of ratio-scaling methods with verbal pain descriptors. Verbal descriptors of pain may provide the best method of scaling different dimensions of the pain experience. Reliable and objective descriptor scales have been developed which separate pain along two dimensions: sensory intensity and affect, or unpleasantness. By using cross-modality matching procedures, specific numerical values can be calculated for each verbal descriptor. These scales have been used to measure the intensity and unpleasantness associated with tooth pulp evoked experimental and clinical pain, and should be extremely useful in the evaluation of acute and chronic dental pain. They will be important experimental and clinical adjuncts in determining the efficacy of non-pharmacological pain control methods such as regional electroanalgesia, biofeedback, relaxation-suggestion and hypnosis.


Contact sensitivity in the pig. II. Induction by intralymphatic infusion of DNP conjugated cells. Pigs were skin painted with the contact sensitizing agent DNFB and afferent lymph was collected for the next 24 h. The lymph cells carried a small amount of DNP and were able to sensitize 40% of homologous recipients. Peripheral blood lymphocytes conjugated with DNFB in vitro to the same degree as afferent lymph cells sensitized 80% of autologous and 40% of homologous recipients: lymphocytes conjugated at higher levels sensitized 75% of animals in each group. Lightly conjugated cells were capable of survival and able to respond to stimulation by mitogens and their ability to sensitize autologous recipients was abolished by heat killing. Highly conjugated cells were not capable of survival, their sensitizing ability was not altered by heat killing and they were able to sensitize incompatible recipients. It is suggested that highly conjugated cells sensitize by a different mechanism which depends on the cooperation of non-lymphocytic cells, not easily mobilized from lymphoid tissue.


Antibody formation to double-stranded DNA in nu/nu and nu/+ mice. Antibody formation to double-stranded (ds-) DNA in nude mice (nu/nu) and normal littermates (nu/+) was studied using a Millipore filter method. The mean antibody titre to ds-DNA in the sera of nu/nu mice was generally higher than that in nu/+ mice raised in the same environment. However, in both genotypes, anti-ds-DNA titres varied according to the environmental condition in which the animals were raised. Mice maintained germ-free showed no significant antibody formation to ds-DNA. Nu/+ thymus and spleen cells passively transferred to nu/nu mice reduced the antibody titre to ds-DNA in the recipients. Immunochemical studies indicated that the antibodies to ds-DNA were IgM. It is suggested that T cell-mediated regulatory mechanisms control antibody formation to ds-DNA and that some infections trigger antibody formation to ds-DNA.


Effects of superoxide dismutase, dithiothreitol and formate ion on the inactivation of papain by hydroxyl and superoxide radicals in aerated solutions. Losses in enzyme activity and sulphydryl content have been studied in aerated papain solutions containing formate, superoxide dismutase and dithiothreitol. Both formate and dithiothreitol converted .OH to .O2-, whereas superoxide dismutase completely suppressed the inactivation by .O2-. Using results from all three systems, the fraction of .O2- reactions with papain that caused inactivation of the enzyme was 0.33 +/- 0.07. The results also showed that the fraction of .OH reactions, which cause inactivation of papain, is significantly higher in aerated than in oxygen-free solutions.


Lymphoid tumours and leukaemia induced in mice by bone-seeking radionucleides. Single intraperitoneal doses of soluble 90Sr and monomeric 239Pu induced generalized lymphomatosis in laboratory mice. Leukaemogenesis due to soluble 226Ra was more uncertain. Clinical expression was variable, but as a generalization the disease was a lymphosarcoma with haematogenous (leukaemic) spread. Only rarely, unlike the commonly recorded forms of natural and X-ray-induced lymphosarcomas, was the thymus apparently the site of onset. The cell-type was lymphoblastic of undifferentiated null form (not T, not B). The average doses of alpha or beta radiation accumulated in the bone-marrow, the presumed site of induction, were at the time of diagnosis usually more than 2500 rad, but, if the cases occurring after radium or low activities of plutonium are accepted as induced, 300-1500 rad of alpha radiation. Mice converted to chimaeras only rarely exhibited any lymphoma, general or local. Abdominal lymphomas were not numerically increased by these radionucleides (perhaps due to shortening of life-span) though some may have been prematurely induced.


Variation in U.V. primary fluorescence-intensity of vital cells depending on 60Co gamma-radiation dose. Using impulse-cytofluorophotometry in the ultra-violet spectral region the author has shown on vital, unstained Ehrlich ascites tumour cells that the primary fluorescence intensity of this tumour is on day 11 after transplantation 20 per cent higher than on day 8. Storage of the vital cells for 25 min at 20 degrees C has no effect on this result. When the cells are exposed to 60Co to gamma-radiation on day 6, a new stable fluorescence level is established afted 20 hours. Measurements of the primary fluorescence intensity depending on dose have shown a significant rise starting from 75 rad at 48 hours after irradiation. The fluorescence intensity rises by 42.5 per cent of the control value at 3000 rad, but only by 31.5 per cent on exposure to 4000 rad.


Alkali-labile sites and post-irradiation effects in single-stranded DNA induced by H radicals. Single-stranded phiX174 DNA in aqueous solutions has been irradiated in the absence of oxygen, under conditions in which only H radicals react with the DNA. It was shown that H radical reactions result in breaks, which contribute approximately 10 per cent inactivation. Further, two types of alkali-labile sites are formed. One is lethal and gives rise to single-strand breaks by alkali and is most probably identical with post-irradiation heat damage and contributes about 33 per cent to the inactivation mentioned above. The other consists of non-lethal damage, partly dihydropyrimidine derivatives, and is converted to lethal damage by alkali. This follows from experiments in which the DNA was treated with osmium-tetroxide, which oxidizes thymine to 5,6-dihydroxy-dihydrothymine. Treatment with alkali of this DNA gives the same temperature dependence as found for the non-lethal alkali-labile sites in irradiated DNA. A similar temperature dependence is found for dihydrothymine and irradiated pyrimidines with alkali.


Fluorescence study of Ehrlich ascites carcinoma cells after X-irradiation. The mechanism of the increase in ultra-violet fluorescence (U.V.F.) intensity of Ehrlich ascites carcinoma cells after X-irradiation were investigated. It was shown that there were two mechanisms: size-dependent (mechanism I) and size-dependent (mechanism II). The basis of mechanism I is most probably an increase in protein content. Mechanism II is related to an increase in the U.V.F. quantum yield, though its biological nature is still unknown.


The effect of X-rays and neutrons on lymphocyte death and transformation. The effects of X-rays and neutrons on human lymphocytes in vitro has been tested. Radiation sensitivity of untransformed lymphocytes was assessed by the appearance of pyknotic cells, and the response of cells after stimulation by phyto-haemagglutinin was tested (a) morphologically and (b) by changes in DNA synthesis, using a labelled thymidine analogue. The data obtained for interphase cells suggest that lymphocytes are a mixed cell population with an insensitive component forming about 20 per cent of the population. The percentage of normal cells observed after both X-ray and neutron irradiation lie on the same dose--effect curve giving an r.b.e. of one. A biphasic response is seen after PHA stimulation with both tests of damage indicating at least two sub-populations of lymphocytes and these give r.b.e. values in the range 1.95 to 2.45. Providing the in vivo response is similar to that in vitro the r.b.e. for damage to circulating lymphocytes will be small and the reduction in white cell count will not therefore be a major factor limiting dose in neutron therapy.


Chromosome aberrations as a dosimetric technique for fission neutrons over the dose-range 0.2--50 rad. An analysis of the literature shows a lack of data about basic biological parameters regarding cytogenetic dosimetry as applied to neutron fields, in the dose range below 50 rad. Human peripheral blood lymphocytes were exposed in vitro to a degraded fission neutron spectrum of mean energy 0.4 MeV. The dose-range was 0.2--50 rad delivered at a dose-rate of about 0.2 or 7 rad/min. The results were processed using a computer programme. Both experimental data for dicentric induction, and theoretical considerations indicate the first-order polynomial as the best fitting dose--response function. The implications of these results for cytogenetic dosimetry are discussed. A comparison with 250 kV X-ray data in the same range gives an r.b.e. of 18.5 +/- 3.1.


Pulse-radiolysis studies of 8-methoxypsoralen. Both eaq- and .OH have been found to react with 8-methoxypsoralen (8-MOP), giving rate-constants of 1.1 X 10(10) M-1 s-1. Transient spectra of products from the reactions of eaq-, .OH with 8-MOP have been characterized. Rate-constants for the oxidation by 8-MOP of reduced and oxidized DNA bases have also been measured and found to lie in the range 3-6 X 10(9) M-1 s-1. Oxidation of reduced bases occurs by electron transfer with 100 per cent efficiency in all cases. However, for oxidized bases, only approximately 25 per cent of the intermediate yield produced by OH attack undergoes electron transfer; the balance of the oxidized base appears to form adducts with 8-MOP.


Effect of salt solutions on the radiosensitivity of mammalian cells. IV. Treatment with NaCl solutions containing ouabain, NEM, PNAP, cysteamine or DMSO. The effects of a wide concentration range of NaCl solutions containing either ouabain, ethanol, para-nitroacetophenone (PNAP), N-ethylmaleimide (NEM), cysteamine or dimethyl sulphoxide (DMSO) on cellular radiosensitivity have been examined. Ouabain and NEM treatment increased the radiosensitivity of V79 Chinese hamster cells, but the action of these chemicals did not depend on the concentration of NaCl. PNAP increased cellular radiosensitivity with increasing NaCl concentration reaching a maximum effect at 0.6 to 0.7 M NaCl. The radioprotective properties of cysteamine, DMSO and ethanol were all strongly dependent on the NaCl concentration in a complex but qualitatively similar manner. DMSO (2.0 M) increased radiation survival of cells after a 1380 rad dose by a factor of about 10(4) when present in 0.075 M NaCl and by a factor of 8.7 when present in 1.2 M NaCl.


An analysis of radiation-induced malignancy based on somatic mutation. If, as has been proposed, the factor which controls the malignant nature of a cell behaves as a recessive genetic character, a somatic mutation in the normal dominant homologous gene of a diploid cell, which carries the malignant factor, may allow the expression of the recessive malignant character. Using this proposal together with the molecular theory of cell survival, a mathematical equation is derived to provide a general description of the dose response for radiation-induced malignancy which is non-linear and peaked. The equation is based on the assumption that a radiation-induced DNA double-strand break can cause a somatic mutation or a chromosome aberration which may leave the cell in a precancerous state. The equation is used to analyse data on the radiation-induced transformation of diploid cells and extended logically to analyse data on the transformation of tetraploid cells.


Blue cone function in a family with an inherited tritan defect, tested with electroretinography and psychophysics. The sensitivity of the blue cone system to low frequency flicker was tested with a psychophysical and an electroretinographical method. With the psychophysical method the subjects, members of a family with an inherited tritan defect, showed no sign of the presence of the blue cone system. With the electroretinogram the sensitivity was also significantly lower than in normal subjects, thus indicating a retinal origin of the tritan defect.


[Severe generalized courses of zoster due to cellular immunologic defects. Importance of an absolute or relative T-cell deficiency]. Four patients with chronic lymphatic leukaemia, M. Hodgkin and metastatic breast carcinoma developed particularly severe generalised herpes zoster, with complications of herpes zoster pneumonia, signs of encephalitis and phrenic nerve paresis. Virus specific complement-fixing antibodies increased regularly or delayed, without strict correlation to the clinical course. However, in all these cases there was a relative or absolute deficiency of T-lymphocytes in the peripheral blood, as a result of the underlying illness and of treatment with cytostatic agents. Because of the vital role of cell-mediated immunity in the control of the varicella-zoster virus (VZV), the observed T-cell deficiency seems to be an important pre-condition for the development of severe generalised herpes zoster.


Growth, luminescence, respiration, and the ATP pool during autoinduction in Beneckea harveyi. The bacterial bioluminescence system is unusual because it is self-induced. In the late logarithmic phase of growth, upon the accumulation of an autoinducer, the synthesis of the components of the system is initiated. We were interested in determining what effect this burst of synthesis and activity has on cellular energy metabolism. The ATP pool of the luminous bacterium Beneckea harveyi was found to dip 10- to 20-fold during the luminescence period, while the respiration per unit cell mass (optical density) increased but by much less. The dip in the ATP pool did not occur in four different types of dark mutants, including one that was temperature conditional and another that was conditional upon added cyclic AMP for luminescence. However, it is neither the synthesis nor the activity of luciferase that is responsible for the ATP dip; the dip does not occur in certain dark "aldehyde" mutants which nevertheless synthesize normal levels of luciferase, whereas it does occur at 36 degrees C in a temperature-sensitive luciferase mutant which forms normal levels of inactive luciferase. Results with other aldehyde mutants implicate the pathway involved in the synthesis of the aldehyde factor with the ATP dip.


The induction of ferritin synthesis in circulating larval red blood cells. The circulating red blood cells formed in bullfrog larvae, chicken embryos, and mouse embryos contain large amounts of ferritin and storage iron in excess of the need for hemoglobin. In contrast, the circulating red cells of adult animals contain little ferritin. Ferritin synthesis and iron storage are coordinated with differentiation and hemoglobin synthesis in the red cells of adults. In order to test the hypothesis that ferritin synthesis could be controlled independently of hemoglobin synthesis and differentiation in the red cells formed early in life, bullfrog larvae were injected with iron to determine if ferritin synthesis was increased in the circulating red cells. Within 17 h after the injection of iron, the synthesis of ferritin, assayed as the incorporation of [14C]leucine by cell suspensions prepared from circulating red cells, was increased from 2.9 to 10.2% of the total protein, and the specific activity of the ferritin synthesized increased from 1100 to 3000 cpm/A280. There was no change in the hematocrit of the animals nor in the specific activity of hemoglobin synthesized by suspensions of red cells (average, 720 cpm/A280). The results suggest that in mature, larval red cells, ferritin synthesis can be controlled by changes in the extracellular environment. The results also indicate that ferritin synthesis can be controlled independently of hemoglobin synthesis with which it is coordinated during erythroid differentiation in adult animals.


A DNase for apurinic/apyrimidinic sites associated with exonuclease III of Hemophilus influenzae. An endonuclease purified from Hemophilus influenzae made single strand breaks in DNA containing apurinic or apyrimidinic sites but had no detectable endonuclease activity on untreated native DNA. The new 5'-termini created at the cleavage sites were base-free deoxyribose 5-phosphate residues. The enzyme preparation also catalyzed the exonucleolytic release of 5'-mononucleotides from bihelical DNA and the hydrolysis of DNA 3'-terminal phosphomonoesters. The phosphatase-exonuclease activity was indistinguishable from that reported by Gunther and Goodgal (J. Biol. Chem. (1970) 245, 5341-5349) and resembled that of exonuclease III of Escherichia coli. The endonucleolytic and exonucleolytic activities could not be separated by electrophoresis, sedimentation, or gel filtration, and they were also affected simultaneously by mutation. The enzymatic activities appear to be functions of a single monomeric protein (M(r) = 30,000).


Intraoperative ureteral trauma: a radiologic approach. Thirty-two patients with inadvertent intraoperative ureteral trauma or ligation are presented. One-third of the injuries were due to general surgical procedures such as lumbar sympathectomy, aortoiliac bypass and colectomy. Because of postoperative pain, discomfort and sedation, these injuries are frequently unrecognized for long periods. This delay in diagnosis results in significant morbidity and mortality. The usual radiologic approach is an excretory urogram supplemented if necessary by a retrograde pyelogram. Temporary percutaneous or operative nephropyelostomies have been employed in seven of these patients.


Growth requirements of calf lens epithelium in culture. The in vitro life span and rate of growth of calf lens cells cultured in serum-supplemented 199 medium can be markedly increased by growing the cells with a layer of mitomycin -killed 3T6 feeders. In the absence of feeders, the epithelial cells are partially blocked in the S period of the cell cycle but show a normal distribution of cells in G1 and G2 when grown with fibroblasts. Increased growth rates and division potential can also be achieved by growing the lens in 199 medium containing 10(-5) M thymidine, and cells grown under these conditions show a normal growth cycle. Our results suggest that lens epithelial cells cultured in medium 199 show a deficient endogenous synthesis of thymidylic acid, and fibroblast feeders or exogenously added thymidine enable them to overcome this deficiency. When grown in the presenct of 10(-5) M thymidine, the lens epithelial cells show a very low serum requirement for cell division in short term culture.


The differential effect of in vivo hydrocortisone on the kinetics of subpopulations of human peripheral blood thymus-derived lymphocytes. The present study was undertaken to determine the effect of in vivo hydrocortisone on the kinetics of subpopulations of normal human peripheral blood (PB) thymus-derived (T) cells. Normal volunteers received a single i.v. dose of hydrocortisone, and blood was taken just before, as well as 4, 24, and 48 h after hydrocortisone administration. T cells were purified from each specimen, and proportions and absolute numbers of T lymphocytes bearing receptors for the Fc portion of IgG (T(.G)) and for the Fc portion of IgM (T(.M)) were enumerated by rosetting T cells with bovine erythrocytes which had been coated with either antibovine erythrocyte IgG or IgM. 4 h after i.v. administration of hydrocortisone, T(.M) cells decreased from 52 (+/-5%) to 23 (+/-6%) of PB T cells (P < 0.01) and the absolute number of T(.M) cells decreased from 1,028 (+/-171) per mm(3) to 103 (+/-23) per mm(3) (P < 0.001). In contrast, relative proportion of T(.G) cells increased from 22 (+/-4%) to 66 (+/-7%), while the absolute numbers of T(.G) cells were essentially unchanged (P > 0.2). In vitro studies involving preincubation of T cells with hydrocortisone before rosette determination of T(.G) or T(.M) cells demonstrated that the decrease in absolute numbers of T(.M) cells did not represent hydrocortisone interference with T(.M) rosette formation, nor did it represent a switch of T(.M) cells to T(.G) cells. Thus, administration of hydrocortisone to normal subjects produces a selective depletion from the circulation of T lymphocytes which possess receptors for the Fc portion of IgM (T(.M) cells) and of T cells which possess no detectable F(C) receptor (T(.non-M, non-G) cells). T(.G) cells are relatively resistant to the lymphopenic effect of hydrocortisone. These data clearly demonstrate that in vivo corticosteroids have a differential effect on the kinetics of identifiable and distinct subsets of cells in the human T-cell class.


Resistance to arteriosclerosis in pigs with von Willebrand's disease. Spontaneous and high cholesterol diet-induced arteriosclerosis. The aortas of 11 pigs (aged 1-3 yr) with homozygous von Willebrand's disease (vWd) were compared with those of 11 normal pigs of the same ages. Six of the controls exhibited multiple arteriosclerotic plaques with intimal thickening of 63-130 mum. In contrast, none of the pigs with vWd had multiple plaques, and only one had a lesion >2 mm in diameter. In a subsequent study, 3-mo-old pigs (11 controls and 7 with homozygous vWd) were placed on a 2% cholesterol diet for up to 6 mo. All of the controls developed arteriosclerotic plaques in the aorta, and in nine of the controls, at least 13% of the entire surface was involved. Intimal thickness ranged up to 390 mum. In contrast, four of the pigs with vWd did not develop such lesions, two developed arteriosclerotic lesions affecting 6 and 7% of the aortic surface, and the seventh had 13% of the aortic surface involved. Most of the pigs with vWd, however, developed flat fatty lesions in contrast to the normal pigs whether on the normal or the high cholesterol diet. There was blue staining of the flat fatty lesions when two pigs with vWd were injected with Evans blue dye antemortem. By electron microscopy, severe endothelial damage was apparent, but there was no intimal proliferation. The coincidence of the impaired platelet-arterial wall interaction and lack of arteriosclerosis in this bleeding disease is discussed.


Suppressor cell-mediated neutropenia in Felty's syndrome. The mechanism of neutropenia in Felty's Syndrome (FS) was tested. The suppressor capacity of mononuclear cells from patients with FS on normal bone marrow granulopoiesis was tested by the in vitro colony forming unit in culture assay. Peripheral blood, bone marrow, and spleen cells from FS patients with marked neutropenia (less than 1,000 neutrophils/mm3) suppressed the colony forming unit in culture of normal bone marrow. Cells from rheumatoid arthritis patients without neutropenia, cells from patients with drug-induced neutropenia without rheumatoid arthritis, or plasma from FS patients failed to suppress the colony forming unit in culture. Though suppressor cells were predominantly thymus-derived (T) cells, monocytes were also effective in suppression. The suppressor efficiency of cells from the various compartments were spleen greater than bone marrow greater than peripheral blood. Splenectomy in FS transiently corrected the neutropenia and eliminated suppressor cell activity. Hyperactive suppressor cells may be responsible for the neutropenia in some patients with FS. Correction of neutropenia in these patients should be directed at modulating the suppressor cell subpopulation.


Purification of specific precipitinogen and extraction of endotoxin from Haemophilus influenzae. After purifying a Haemophilus influenzae precipitinogen from endotoxic activity by means of ultracentrifugation, column chromatography (Sepharose 6B) and ion exchange chromatography (DEAE Sephadex A25) a fraction was obtained which still contained a specific precipitinogen that was virtually free of endotoxin. Furthermore, during the chromatographic procedures fractions with a high and a low molecular weight endotoxic activity were found. The limulus lysate test was more sensitive in the high molecular weight fractions and the LD50 in mice in the low molecular weight fractions with endotoxic activity.


Topographic projections between the nucleus isthmi and the tectum of the frog Rana pipiens. The connections between the nucleus isthmi and the tectum in the frog have been determined by several anatomical techniques: iontophoresis of horseradish peroxidase into the tectum, iontophoresis of 3H-porline into the nucleus isthmi and the tectum, and Fink-Heimer degeneration staining after lesions of the nucleus isthmi. The results show that the nucleus isthmi projects bilaterally to the tectal lobes. The ipsilateral isthmio-tectal fibers are distributed in the superficial layers of the tectum, coincident with the retionotectal terminals. The contralateral isthmio-tectal fibers travel anteriorly adjacent to the lateral optic tract and cross the midline in the supraoptic ventral decussation, where they turn dorsally and caudally; upon reaching the tectum, the fibers end in two discrete layers, layers 8 and A of Potter. The tectum projects to the ipsilateral nucleus isthmi and there is a reciprocal topographic relationship between the two structures. Thus, a retino-tecto-isthmio-tectal route exists which may contribute to the indirect ipsilateral retinotectal projection which is observed electrophysiologically. The connections between the nucleus isthmi and the tectum in the frog are strinkingly similar to the connections between the parabigeminal nucleus and the superior colliculus of mammals.


Lymphocyte subpopulations in maintenance hemodialysis patients. Lymphocyte counts and T,B and null cell proportions were analyzed in forty non-splenectomized, non-nephrectomized maintenance hemodialysis patients, and compared with those in controls. The patients were lymphopenic, but had normal T, B and null cell percentages, regardless of tehir primary renal disease. It is postulated that their lymphocyte suppression is non-selective and may be related to the state of controlled azotemia or to the dialysis procedure.


Successful coronary artery bypass in hemodialysis patients. Cardiovascular disease is the major casue of death in maintenance hemodialysis patients. We report two chronic hemodialysis patients with unstable, disabling angina in association with severe coronary artery disease involving all three major vessels. Both successfully underwent coronary artery bypass surgery and subsequently experienced dramatic clinical improvement. Principles of management are discussed and it is suggested that hemodialysis patients should not be arbitrarily denied consideration of coronary artery bypass surgery when it is otherwise indicated.


T lymphocyte-mediated cytolysis: dissociation of the binding and lytic mechanisms of the effector cell. To determine functional relationships between the cytotoxic T lymphocyte (CTL) receptor for target binding and the lytic mechanism, we have studied the reaction between two immunized CTL populations (AalphaB and BalphaA), both at the population and the single-cell level. When studied at the population level, the reaction of AalphaB with BalphaA (bidirectional system) resulted in formation of AalphaB/BalphaA conjugates and bidirectional cytolysis. However, when the viability of cells in individual AalphaB/BalphaA conjugates was analyzed, unidirectional instead of bidirectional lysis occurred. These results indicate that under conditions that are conducive to lysis, binding of a potentially lytic cell to its target does not necessarily result in target lysis. Short heat treatment of CTL (44 degrees C, 10 min) totally abolished their lytic activity, without affecting their capacity to bind specifically, thus dissociating the binding from the lytic activity of the CTL. The cytolytic activity is probably associated with, or triggered by the CTL-binding unit. The binding unit, on the other hand, appears to be a functional receptor of the CTL, which is involved in but not sufficient to bring about lysis.


Role of T lymphocytes in adjuvant arthritis. II. Different subpopulations of T lymphocytes functioning in the development of the disease. The effects of treatments with cyclophosphamide (CY), hydrocortisone and anti-thymocyte sera (ATS) on the development of adjuvant arthritis (AA) were examined in WKA rats inoculated with wax D to induce AA. A single injection of 25 to 50 mg/kg of CY given 2 to 3 days before wax D inoculation caused severe arthritis with high incidence, whereas larger doses of CY were less efficient. Furthermore, the enhancing effect of CY pretreatment was abolished by passively transferred normal syngeneic thymocyreated with ATS and guinea pig C. On the basis of these results and the previous observations on the effect of adult thymectomy and low-dose irradiation, we concluded that this enhancing effect of CY pretreatment was caused by selective depletion of suppressor T lymphocytes. Pretreatment with 12.5 mg of hydrocortisone also caused severe arthritis. This enhancing effect of hydrocortisone could also be due to the elimination of those suppressor cells. In contrast, in vivo pretreatment with ATS showed striking inhibition on the development of AA, suggesting that ATS could eliminate T lymphocytes that were responsible for eliciting this disease. Thus it appears that at least two T cell subpopulations are involved in the development of AA, one is an ATS-sensitive T2 subpopulation that is effective for induction of AA and the other is a T1 subpopulation that regulates this disease process.


Inactivation of antigen-responsive clones with antisera specific for IgM or IgD. The present study examines the isotype of cell surface immunoglobulins involved in triggering a response to thymus-dependent and thymus-independent antigens. Antibodies to immunoglobulin isotypes were used to block the in vitro interaction of receptor IgD and IgM with antigen. The results indicate that both IgD and IgM are necessary to trigger a response to TNP-SRBC but that only IgM is required for responsiveness to TNP-Brucella. Limiting dilution studies indicate that the inhibition of the immune response by antibody occurs at the level of precursor activation and suggest that there is no effect of antibody on subsequent proliferation of the daughter cells.


The analysis of the monoclonal immune response to influenza virus. III. The relationship between stimulation of virus-primed precursor B cells by heterologous viruses and reactivity of secreted antibodies. Individual splenic precursor B cells from BALB/c mice primed with influenza virus PR8[A/PR/8/34 (H0N1)] were stimulated in vitro in the splenic fragment culture system by homologous or various heterologous influenza viruses. The specificity of the stimulated precursor cells was determined by analysis of the antibodies secreted by the ensuing plasma cell clone in a radioimmunoassay (RIA). Viruses of the H2N2 and H3N2 subtypes were unable to stimulate hemagglutinin (HA)- or neuraminidase (NA)-committed precursor B cells but did efficiently stimulate chicken host component (ChHC)-committed precursors. Viruses of the H1N1 and H0N1 subtypes could stimulate precursors committed to any of the three viral surface components. Analysis of the fine specificity of HA-committed B cells showed that BEL(H0N1) and CAM(H1N1) stimulated almost exclusively precursors whose clonal antibody product reacted with the stimulating virus in the RIA. On the other hand, WSE and MEL (both H0N1) quite frequently were able to stimulate precursors whose clonal antibody product did not react with the stimulating virus in the RIA. These results suggest that the stimulatory interaction of viruses with the cell-bound immunoglobulin receptors is slightly less affinity dependent than the antigen-antibody interaction in the RIA.


The kinetics of target cell lysis by cytotoxic T lymphocytes: a description by Poisson statistics. The kinetics of T cell killing are analyzed with the assumption that encounters between killer and target cells occur at random. The application of Poisson statistics leads to a number of theoretical predictions which were then tested experimentally. Good agreement between data and theory was found.


Suppression of the mitogen-stimulated blastogenic response during reticuloendotheliosis virus-induced tumorigenesis: investigations into the mechanism of action of the suppressor. Splenic lymphocytes from chickens infected with reticuloendotheliosis virus (REV) are suppressed in their ability to undergo PHA-induced blastogenesis. This suppression can be detected within 72 hr after virus injection and requires active viral infection since i.p. or i.v. injection of UV-inactivated REV does not result in inhibition of the blastogenic response. Suppressor cells from the spleens of REV-infected birds severely inhibit the ability of spleen cells from uninfected chickens to respond to PHA at a ratio of 1:20, suggesting that each suppressor cell may be capable of suppressing more than one target cell. Contact between suppressor and target cells is required for the rapid inhibition of the normal PHA response. The suppressive mechanism is cytostatic in nature, and apparently of host origin since neither REV, nor REV-infected (or transformed) cells mediate the suppression. The ability of the suppressor cells to impair the blastogenic response of spleen cells is sensitive to trypsin, suggesting that an inhibitory protein is exposed on the surface of the suppressor cells.


Suppressor cell activity in tumor-bearing mice. I. Dualistic inhibition by suppressor T lymphocytes and macrophages. Spleen cells from mice bearing methylcholanthrene-induced fibrosarcomas impaired mitogen responses of normal syngeneic lymphocytes. Nylon wool column and other depletion techniques were utilized to characterize the cellular source of suppressive activity in tumor-bearing host (TBH) spleens. Evidence is presented for two distinct suppressor cell systems operating in the spleens, but not lymph nodes, of BALB/c mice bearing transplanted tumors. Spleens from TBH were shown to have greatly increased numbers of macrophages over their normal counterparts. TBH macrophages were observed to have suppressive activity at low in vitro concentrations. Anti-Thy 1 serum treatment of TBH macrophages abrogated low dose inhibition but not suppression due to high numbers of macrophages. No functional difference was detected between anti-Thy 1 serum-treated TBH and normal splenic macrophages. In a macrophage-depleted culture system, mildly nylon wool adherent, anti-Thy 1 serum, and hydrocortisone succinate-sensitive suppressor cells could be detected. Soluble supernatant products of TBH spleen and thymus cells were also found to inhibit in vitro mitogen responses, whereas TBH macrophages and lymph node cells demonstrated no soluble suppressive activity. The major source of soluble inhibitor of DNA synthesis (IDS) seems to be an anti-Thy 1 serum, hydrocortisone-sensitive population.


Imbalances in T cell subpopulations in multiple sclerosis patients. Abnormal proportions of a distinct T cell subpopulation able to bind IgG immune complexes (T.G cells) were found in peripheral blood samples from patients with MS. About 50% of the patients examined had an overabundance of T.G cells. The possible role of these cells in the pathogenesis of MS is considered.


Isolation and characterization of a rabbit T lymphocyte-specific antigen. Rabbit T lymphocytes may be differentiated from B lymphocytes by the presence of a T lymphocyte-specific surface antigen. This unique antigen has been extracted from the plasma membrane of rabbit thymocytes by 3 M KCl. The presence of the antigen in the membrane extract was demonstrated by inhibition of cytotoxicity with goat anti-rabbit T cell serum (ATS). The crude membrane extract was fractionated by gel electrophoresis and the fractions containing the T cell antigen identified by inhibition of cytotoxic ATS and by passive hemagglutination. The purified T cell antigen was found to have a m.w. of approximately 12,000 and contained approximately 2.5% carbohydrate. Evidence was also obtained to suggest that the rabbit T cell antigen exists in multiple forms, each having the same m.w. but exhibiting different electrophoretic characteristics.


Clq binding activity in lupus erythematosus: correlation with the "lupus band test". A 131I C1q binding assay was employed to estimate C1q binding activity (C1q BA), presumably due to the presence of circulating immune complexes, in lupus erythematosus sera. Thirteen of 16 sera from patients with systemic lupus erythematosus (SLE) had elevated C1q BA. Only 1 of 17 sera from patients with generalized discoid LE (GDLE) and none of 5 localized DLE patient's sera had elevated C1q BA. A correlation between a positive LE band test and the level of C1q BA was apparent. A positive LE band test tended to occur in patients with higher levels of C1q BA, whereas a negative LE band test tended to occur in patients with low levels of C1q BA.


Antibodies to nuclear antigens in patients with renal failure. Nuclear material, presumably from damaged leukocytes, adheres to hemodialysis membranes. Previous studies have shown an increased prevalence of antibodies to nuclear antigens in chronic dialysis patients. Present studies verify the increased prevalence of antibodies to nuclear antigens in 52 of 243 chronic dialysis patients (21.4%). Because antibodies are present only intermittently, prevalence increases with repetitive blood sampling. In patients tested every 3 months over several years on chronic dialysis, the prevalence of antinuclear antibodies approaches 100%. The present studies also demonstrate, however, that the prevalence of antibodies is increased in patients with renal failure who have never undergone hemodialysis (10 of 86, 11.6%). Thus the tendency to form antibodies to nuclear antigens may be associated with renal failure rather than dialysis per se. Higher prevalences with increased time on dialysis and transplantation failure may accordingly reflect the greater duration of renal failure. The prevalence of antibodies to nuclear antigens was not significantly influenced by age, sex, type of kidney disease, major blood group, HLA tissue type, or coil reuse. Significantly lower hematocrits and white blood counts were noted when antibodies were present.


Distribution of alpha 1-antitrypsin in normal, granuloma, and tumor tissues in rats. Purified rat alpha 1-antitrypsin and albumin were radiolabeled, and their distribution and catabolism in various tissues were studied in rats with inflammatory granuloma or transplanted sarcoma. Intravenously administered labeled alpha 1-antitrypsin accumulated remarkably in extravascular spaces of the granuloma or sarcoma tissues. Among the normal organs examined, the lung preferentially incorporated alpha 1-antitrypsin. Furthermore, most of the alpha 1-antitrypsin accumulated in these three tissues remained in a TCA-precipitable form throughout the observation period. Since alpha 1-antitrypsin is incorporated in large amounts into inflammatory or tumor tissues, it could play a role in regulation of inflammatory processes or in controlling the proliferation of a tumor. The studies on TCA fractionation also suggest that liver and kidney provide the main sites for degradation of this protein. Although the accumulation of labeled albumin in granuloma and sarcoma was less marked, it showed essentially the same distribution and degradation pattern as alpha 1-antitrypsin in both morbid states.