[The spleen in "nude" mice: an immunological and immunocytochemical study (author's transl)]. An immunological and immunocytochemical study to compare spleen cells of Nude (C57B1) and Swiss mice is reported. The percentage of surface immunoglobulins bearing lymphocytes is identical in both strains of animal (40%). An activated C3 receptor is present in the majority of these cells and has been demonstrated within the germinal centers in both cell suspensions and frozen tissue sections. The population of Fc receptor bearing cells is more heterogenous, but the percentage of these is nearly identical in both Swiss and Nude mice. T-cell specificity is identified with an anti-thymocyte and an antibrain antiserum using an indirect immunoperoxidase method. The percentage of cells detected with the anti-thymocyte antiserum is very low in the spleen of Nude mice (5%) compared to that in the Swiss mice (42%). In the Nude mice, the anti-brain antiserum detects up to 30% of the spleen cells which may be considered as precursors and, so far, these 30% of cells in the Nude mice are considered as "null" cells, compared to only 20% in normal mice. The localization of the various immunocompetent cells within the spleen tissue is determined by immunofluorescence, immunoenzymology and the EAC rosette test. This study leads to the conclusion that in the periarteriolar areas of the spleen of the Nude mice, immune cells are sparse but that thymo-independent cells are located in the same region in both species of mice.


Nutrition of Bacillus brevis ATCC 9999, the producer of gramicidin S. The nutrition of the gramicidin S (GS) producer Bacillus brevis ATCC 9999 was studied with respect to growth and antibiotic production. In a complex medium containing yeast extract and peptone, only glycerol, d-fructose, and meso-inositol contributed to growth, whereas other hexoses, pentoses, polyols, polysaccharides, and organic acids were inactive. Glycerol and fructose were inhibitory to GS formation; inositol had no effect. In chemically defined media containing a mixture of five amino acids (glutamine, methionine, proline, arginine, and histidine) necessary for good growth, again only glycerol, d-fructose, and meso-inositol served as effective carbon sources for growth. With respect to antibiotic formation, inositol was much poorer than glycerol or fructose. In the presence of glycerol and the five amino acids, addition of 0.1% l-phenylalanine specifically stimulated GS production. Growth kinetics in this defined medium showed two phases: early and rapid growth at the expense of the amino acid mixture, followed by glycerol utilization after a diauxic lag of 2 to 3 h; inositol utilization suffered a much longer lag period. GS formation was initiated after the diauxic lag. Ammonium sulfate was the best individual nitrogen source, but the combination of the five growth-stimulatory amino acids was more potent for GS production. The level of potassium phosphate influenced GS production. High levels stimulated the growth rate but progressively inhibited GS production. Methionine was the best sulfur source. A defined medium (F3/6) that supported rapid exponential growth (mu = 0.53 h(-1)) followed by progressively slower growth over a 30- to 40-h period was devised containing fructose, six amino acids, and mineral salts. GS formation was initiated as the cells left the exponential growth phase and reached high levels (1,500 mg of GS per liter; 0.19 mg of GS/mg of dry cell weight) after 40 to 50 h of incubation.


[Affinity of concanavalin A for "trypanosoma equiperdum": purification of the antigenic type specific glycoprotein fraction (author's transl)]. Binding of concanavalin A by different clones of Trypanosoma equiperdum was checked by its ability to agglutinate living trypanosomes. The clones differ by their agglutinability, showing differences in the distribution of the binding sites. However, the property of concanavalin A to form precipitates with the crude antigen preparations of all the clones studied, allowed to isolate by affinity chromatography the glycoprotein fraction responsible for the antigenic variation and to enter upon its physicochemical study.


"5-Amino-5"-deoxybutirosin, a semisynthetic analogue of butirosin A: antibacterial activity in vitro and in mice. Aminodeoxybutirosin (AD-BTN), the 5''-amino-5''-deoxy derivative of butirosin (BTN), was synthesized to improve on the antibacterial activity of BTN by preventing bacterial enzymatic phosphorylation at the 5'' position. AD-BTN possesses the spectrum characteristic of BTN and gentamicin (GTM) and was active at low levels in vitro against a wide variety of gram-negative species including Pseudomonas aeruginosa, indole-positive Proteus and Serratia marcescens; its action was bactericidal against both light and heavy inocula, and it was not antagonized by human serum. AD-BTN was as active as GTM against GTM-sensitive P. aeruginosa in vitro and in mice, and was markedly improved over BTN. AD-BTN retained the good activity of the parent compound against other gram-negative pathogens. Whereas GTM minimal inhibitory concentrations were elevated 35-fold against GTM-resistant P. aeruginosa in vitro, the minimal inhibitory concentration of AD-BTN was only doubled. At 6.3 mug/ml, AD-BTN inhibited 68% of 82 isolates insusceptible to that concentration of GTM. In murine toxicity tests AD-BTN was about one-third to one-half as toxic as GTM.


Type C oncornavirus isolation studies in systemic lupus erythematosus. I. Attempted detection by isopycnic sedimentation of 3H-uridine-labelled virions. Type C oncornavirus isolation was attempted from cell cultures of tissues from 7 patients with systemic lupus erythematosus. Detection was based on the characteristic sedimentation of 3H-uridine-labelled virions at a density of 1-16 g/ml. Cultures positive by this method were negative by two other criteria for type C viruses: characteristic virions by electron microscopy and the viral enzyme RNA-directed DNA polymerase. The positive results were probably due to cellular damage by prolonged radiolabelling, with release of organelles containing labelled RNA sedimenting at the same density as type C viruses.


[Humoral immunity and blastic transformation of lymphocytes in trisomy 21]. Studies of immunologic parameters in 52 children with trisomy 21 revealed that -the changes in serum immunoglobulin patterns were minimal, although some delay in the age-dependent rise of IgG concentration was observed; -lymphocyte reactivity to phytohemagglutinin showed a shift with maximal 3H-thymidine incorporation at low PHA concentration; -presence of HBs antigen at an increased incidence, possible existence of anti-thyroid antibodies in serum of trisomy patients and also in that of their parents, abnormalities in phagocytosis, illustrate that trisomy 21 can be associated with multiple immunologic dysfunctions.


[Immunochemical investigations on the protein of the "pregnancy zone". XI. Serum concentration of the pregnancy-associated alpha2-glycoprotein (Pregnancy Zone protein) in normal pregnancy (author's transl)]. A quantitative study of the pregnancy zone proteins was made in the sera of 383 healthy pregnant women by means of radial immunodiffusion according to Mancini and co-workers. The serum levels related to a pregnancy serum standard were measured from the 6th to 44th week of pregnancy. The serum concentration of the protein showed a considerable individual variation. The mean concentration began to rise in gestational weeks 8 to 24. Until the week 32 a rather stable average level was reached. Before delivery a slight decrease was observed. Women with over term pregnancies showed particular high mean concentrations. The differences of serum levels were statistically significant until the gestational week 14.


On the occurrence of the pregnancy zone protein (PZ) in gynecological cancer. Reports have been made that the pregnancy zone protein (PZ) is found in increased amounts in sera of patients with malignant disease. The serum concentration of PZ was measured by a radioimmunoassay in 83 women with the diagnosis of gynaecological cancer or other malignancies. Values did not significantly differ from values previously recorded in apparently healthy women. There was no support for previous data of increased levels of PZ in cancer patients, when the age-dependent increase of this serum factor was considered.


Alterations of the myoid pericanalicular layer in liver. A light microscopical pilot study of human autopsy material. Previous histochemical studies demonstrated a myoid layer along biliary pathways. This report describes alterations of the pericanalicular layer in various lesions. Carnoy- or methacarn-fixed sections of livers from 200 autopsies were treated with the tannic acid-phosphomolybdic acid-dye method for myosins. In other series, methods for myosins were combined with the picro-Sirius Red F3BA stain or the Prussian Blue reaction. In normal liver, bile canaliculi were lined by a narrow myoid layer. In other cases, eg, hepatic cirrhosis, the pericanalicular layer showed slight to moderate thickening. In cases with obstruction or atresia of bile ducts, the dilated canaliculi were surrounded by a prominent myoid layer. These observations support theories that the pericanalicular layer is contractile and promotes bile flow under positive pressure.


[Structural-reactive properties of interoceptors (intravital study)]. It has been shown that between living and surviving receptors of a central and local origin there is similarity in a number of morphological, reactive and tinctorial properties as well as in their response to tissue fixation. At the same time these structures possess a number of specific properties making them different from one another.


[Mast cells as regulators of tissue homeostasis and their place in the series of biological regulators]. The most essential features of the mast cell are continuous production (and partially also absorption from the environment), deposition, and secretion of the two groups of biologically active substances of the regulatory type: heparin and other acid clycosaminoglycanes, and also histamine and, probably, other biogenic amines. These substances are antagonists and have an influence on the homeostasis of the microregion (capillary-connective tissue-parenchyma) -- the main area of action of the mast cells. The mast-cell population is characterized, as compared with other cellular populations of the microregion, by a smaller size, greater morphological and functional diversity, functional duality (capability of producing both a positive and negative effect), and therefore possesses a number of essential properties the biological regulator should meet. The system of mast cells differs from the regulators of a higher order (the nervous and endocrine system) by a smaller radius of action and a greater simplicity of the structure, the latter manifests itself in equality of all its components. The conclusion is drawn that mast cells may be considered as the regulators of tissue homeostasis and a last link in the general reaction of adaptation at the cellular level.


Herpetic corneal epithelial disease.

Herpetic corneal epithelial disease. The clinical differentiation of corneal epithelial lesions due to herpes simplex or herpes zoster may be confusing. Practical clinical tests, including the use of topical ocular stains, are useful to differentiate corneal epithelial lesions caused by these two viruses. Two distinctive types of zoster corneal epithelial disease may be seen; an early dendritic form, and a delayed form characterized by corneal mucus plaques that may take a dendriform pattern. These plaques are composed of mucus that is adherent to swollen, degenerating epithelial cells. The clinical differentiation between these two viruses is essential since topically applied corticosteroids are contraindicated in epithelial herpes simplex and often are indicated in the management of epithelial herpes zoster.


An assessment service for country children. An assessment service for country children is described, which is characterised by its joint staffing by the Health Commission and Department of Education, and by its multidisciplinary approach to assessment. Remediation is also provided when indicated, and an increasingly comprehensive follow up service is developing in conjunction with the Consultative Aerial Health Service.


Some immunological aspects of a recent Australian isolate of infectious bronchitis virus. An infectious bronchitis virus, designated G48, isolated from birds during an outbreak of nephritis in a previously vaccinated broiler flock, overcame the resistance induced in birds vaccinated with 2 commercially available vaccines. Birds vaccinated with the A isolate of infectious bronchitis resisted challenge with this new virus. Cross neutralisation studies revealed that the new virus was serologically distinct from the 4 viruses tested. Homologous antiserum to G48 did not neutralise the other viruses and only antiserum to the A virus completely neutralised the new virus.


Interaction of cell-membrane prolactin receptor with its antibody. Antisera against a partially purified prolactin-receptor preparation derived from pregnant-rabbit mammary glands were generated in guinea pigs. On double immuno-diffusion, each antiserum produced a single precipitin line with the prolactin receptors. The anti-receptor sera also specifically inhibited the binding of 125I-labelled sheep prolactin to membrane particles as well as to highly purified prolactin receptors derived from the rabbit mammary glands. The same antisera, however, had no effect on the binding of 125I-labelled insulin to the same membranes. These antisera did not bind or destroy prolactin. Moreover, the binding of 125I-LABELLED PROLACTIN TO MEMBRANE PARTICLES DErived from different tissues from a number of species was also inhibited by the antisera, thus suggesting that the immunological determinants of the prolactin receptors are similar in various tissues derived from different species. The factors in the antisera that were responsible for inhibiting the binding of 125I-labelled prolactin to its receptors were found to be associated with the gamma-globulin fraction. In addition, 131I-labelled gamma-globulins derived from one antiserum were shown to bind to membrane particles derived from mammary glands, and an increase in binding of gamma-globulin was accompanied by a decrease in binding of prolactin. Kinetic analyses of inhibition of 125I-labelled prolactin binding by antisera by using the methods of Lineweaver & Burk [J. Am. Chem. Soc. (1934) 56, 658-666] and Dixon [Biochem. J. (1953) 55, 170-171], revealed that the mechanism is a hyperbolic competitive inhibition. The demonstration of hormone-receptor-antibody complexes further favours this mechanism. The availability of anti-receptor sera should facilitate studies on the functional role as well as other biochemical, immunological and physiological properties of the prolactin receptors.


Histological arguments for collagen and elastin synthesis by primary cultures of rat aortic media cells. Using the histological staining methods of Weigert and of Masson on primary cultures of rat aortic media cells, we obtained additional proofs of the smooth muscle cell's ability to secrete collagen and elastin in vitro: the percentage of positive flasks with aorta rings was the same throughout the follow-up, but increased gradually for the new tissue growing around the rings.


The effect of the duration of cholesterol feeding on the development of sudanophilic lesions in the rabbit aorta. Polar coordinate mapping was used to determine the rate of growth of individual sudanophilic lesions on the aortic wall around several major branches of the aortae of cholesterol fed rabbits. Four groups, with 6 8-month old male albino white rabbits in each, were used in the study. One group served as a control and the remaining 3 were fed a diet of 2% cholesterol and 6% heated corn oil mixed with ground rabbit pellets for 4, 8, and 10 weeks each. Animals were sacrificed, the aortae removed, stained with Sudan III, pinned at in vivo dimensions, and mapped by the polar coordinate method. No sudanophilic lesions were observed in the control animals. In the experimental groups, the early lesions, except the coronaries, were almost entirely distal to the orifices, and maintained roughly the same contour while spreading around the orifice. The coronary lesions completely encircled the orifices as described previously. As lesions progressed, they became elevated and often granular, so that the lesions themselves may have affected flow profiles around the orifices. Lesions around adjacent orifices were fused in 48% of the cases after 10 weeks on the diet, as opposed to 2% after 4 weeks on the diet. More prolonged experiments were not possible with this diet as the animals developed jaundice and diarrhea. Hemodynamically, these results suggest that early sudanophilic lesions in cholesterol-fed rabbits develop on the aortic wall in areas of high shear stress.


In vitro synthesis of "amyloid"fibrils from insulin, calcitonin and parathormone. Insulin, calcitonin and parathyroid hormone subjected to one of two procedures-acidification and heating or incubation with mouse kidney lysosomal extracts-assumed a nonbranching fibrillar structure, 7 to 10 nm in diameter. The preparations showed green birefringence after Congo red staining. The in vitro synthesis from different hormonal polypeptides of fibrils, fulfilling the criteria for the identification of amyloid, indicates that these criteria are related to conformational rather than to compositional properties, and suggests that these hormones may provide the subunit of the amyloid formed in the corresponding endocrine organs.


The sunburn cell in mouse skin: preliminary quantitative studies on its production. Quantitative studies on sunburn cell (SBC) production in mouse epidermis are reported. Ultraviolet radiation (UVR), 260-300 nm, produced a dose related effect with respect to SBC formation. Maximal spectral reactivity was at wavelengths shorter than 300 nm. In 8-methoxypsoralen (8-MOP) photosensitization, SBCs were also produced and this response is dose related with respect to long wave UVR dose. In fluorescein photosensitization, provoked by 487 nm light, SBCs were not produced. The response in 8-MOP photosensitization and 254 nm UVR (given alone) differed in respect of time course, but it is considered that both may perhaps to initiated by a DNA lesion.


Labetalol in long-term treatment of hypertension. 1 Labetalol, a beta- and alpha-adrenoreceptor-blocking drug, has been used in the treatment of hypertension in a total of 32 patients for over 4.5 yr, 14 patients for 3 yr or more. 2 Labetalol seems to have a similarly potency of methyldopa and the adrenergic neurone-blocking drugs. 3 Postural hypotension was only observed at doses over 2 g/d. It was the reason for stopping treatment in two patients. 4 Dosage varied; average 889 mg/d, range 75-3,200 mg. Tolerance did not develop. 5 Side-effects led to drug withdrawal in four patients


Terminal deoxynucleotidyl transferase measurements in the differential diagnosis of adult leukaemias. Terminal deoxynucleotidyl transferase (TDT) is an unusual DNA polymerase that does not use template information to synthesize new strands of DNA. It is normally found in high concentration in thymus (50 u/10(8) cells) and in low concentration in bone marrow (less than 5 u/10(8)). We report TDT measurements in the marrow and/or peripheral blood of 51 adult patients, 28 of whom had leukaemia. TDT is present in very high levels (greater than 50 u/10(8) cells) in leukaemic lymphoblasts and in low levels in leukaemic myeloblasts (less than 9 u/10(8) cells). Of two patients who developed lymphosarcoma-cell leukaemia following treatment of poorly differentiated lymphocytic lymphoma, one had high and one low levels of TDT in the leukaemic blast cells. Leukaemic cells from three of seven patients with chronic myeloid leukaemia in blast crisis had TDT levels within the range expected of acute lymphoblastic rather than acute myeloid leukaemia. High TDT in leukaemic cells probably marks them as derivatives of lymphoid progenitor, thymic or pluripotential stem cells. Quantitative assay of TDT may provide information useful in classifying haematological neoplasms.


Amniotic fluid levels of fibrin(ogen) degradation fragment E and alpha-fetoprotein in normal pregnancy and with fetal neural tube defect. Amniotic fluid levels of alpha-fetoprotein (AFP) and fibrin(ogen) degradation fragment E (FgE) were measured in 214 normal subjects and 27 pregnancies associated with an abnormal fetus (open neural tube defect or exomphalos). AFP levels showed no overlap between the normal and abnormal groups, thus confirming the reliability of the AFP assay in the detection of these abnormalities. FgE levels however showed considerable overlap and could not be used as a diagnostic parameter. Agarose gel chromatography of the amniotic fluid revealed the presence of large molecular weight FgE related antigen in open neural tube defect suggesting that leakage of proteins from exposed capillaries is responsible for the elevation of amniotic fluid protein levels.


The significance of raised maternal serum alpha-fetoprotein levels. A radioimmunoassay for alpha-fetoprotein (AFP) is described and normal ranges for both maternal serum and in amniotic fluid throughout pregnancy are defined. Maternal serum AFP levels in at risk pregnancies were found to be no different from those in normal pregnancies. AFP levels in pregnancies complicated by neural tube and other congenital defects, fetal death or maternal hypertension are documented. Eight patients with a fetus deformed by anencephaly or an open spina bifida were tested before 22 weeks; seven of them had raised serum AFP levels. Other causes of raised serum AFP levels are described and the significance of a raised serum AFP level is discussed with particular reference to screening programmes.


Anencephalic conjoined twins.

Anencephalic conjoined twins. A case of anencephaly in female thoracophagus conjoined twins is presented and discussed.


Structural and topological homology between porcine intestinal and renal brush border aminopeptidase. A method for the preparation of closed, right-side-out vesicles from the brush border membrane of the kidney proximal tubules is described. The aminopeptidase known to be bound to this membrane was investigated in order to compare its properties with those already reported for the intestinal enzyme. Both are composed of a hydrophilic, catalytically active part lying on the external side of the membrane and a short hydrophobic domain probably located in the N-terminal region of one of the subunits ensuring fixation to the lipid matrix. The enzyme were also found to be clinically similar. Moreover, a quantitative immunological technique showed that they contained 6 cross-reacting determinants, consistent with a very high degree of homology. Four of these determinants were accessible in the bound form of the enzymes in the region of the active site. The other two, probably related to the junction between the hydrophilic moiety and the hydrophobic anchor were completely masked in the bound form. The remainder (6 in the intestinal and 4 in the renal enzyme), were heterologous. The accessibility of two well determinants in this latter group was substantially reduced, perhaps by the proximity of the lipid and/or of other enzyme molecules.


Alpha-globulins in the surveillance of colorectal cancer. The alpha1 and alpha2-globulins have been studied at various stages in the evolution of colo-rectal cancer. The alph2 was elevated in some primary tumours and rose in metastatic cancer especially when it involved the liver. Some apparently tumour free patients had an unexplained elevation of alph2-globulins. The macroglobulins were not a major constituent of the raised alpha2-globulins. Haptoglobulin levels were found to be a useful indicator of tumour activity, when their level was raised in metastatic cancer it was usually with an antecedent or coincidental rise of plasma CEA. Primary tumours may cause a high haptoglobin response without an elevation of CEA.


Endogenous DNA polymerase of a transformation-defective rous sarcoma virus: characterization and comparison with the enzyme of the non-defective parent. An RNA-directed DNA polymerase associated with transformation-defective (td) segregant of Rous sarcoma virus (RSV) has been characterized. The enzyme required both a monovalent and a divalent cation, a sulfhydryl reducing agent, and all four deoxyribonucleoside triphosphates for the expression of maximal activity. Sensitivity of the endogenous RNA-directed DNA polymerase activity to a low concentration of pancreatic RNase indicated that the enzyme utilized the td virus endogenous RNA as template. Maximal DNA synthesis was observed in a reaction mixture of pH 8 - 8.5 at 45 C with a manganese concentration of 1 mM. The enzyme of the td virus responded to exogenous template-primers in a manner characteristic of DNA polymerase of RNA tumor viruses, and the response became substantially greater when noncomplementary precursors were omitted from the reaction mixture. The endogenous reaction kinetics were examined. Three phases of DNA synthesis could be distinguished. Evidence was obtained showing that during the third and slowest phase of DNA synthesis the reaction mixture was not depleted of precursors and that the enzyme was fully active to initiate DNA synthesis with newly-added viral or synthetic RNA templates. Comparison of TMP and dAMP incorporation kinetics suggested that at the initial phase the enzyme preferentially copies A-rich region(s) of viral RNA. A comparison was also made between the endogenous reaction of the td virus and that of its parent sarcoma virus. The pH optimum, metal ion requirements, effect of sulfhydryl agents, response to exogenous template-primers, and kinetics of DNA synthesis, were all compared. No significant difference between the reaction of the td virus and its sarcomatogenous counterpart could be demonstrated.


Cis AB blood groups. Immunologic, thermodynamic and quantitative studies of ABH antigens. Fifteen samples of cis AB bloods belonging to six unrelated families were tested by serological and thermodynamic assay techniques. The B and H antigens of cis AB bloods differ significantly from those of trans AB bloods. Differences were found among unrelated samples, but identical results were obtained within a given family : this could mean that there had been as many mutations as there were families.


[Experimental teratocarcinoma in mice: a model system for the study of the relationship between cellular surface antigens and embryonic differentiation]. Several cell lines (either of embryonal carcinoma or of differentiated cells derived from teratomas) have been established in vitro from transplantable testicular teratomas. Primitive cell lines, propagated in vitro as embryonal carcinoma have retained the ability of the original tumor to differentiate in vivo or in vitro into must embryonic cell types. Features of this model system for study of early embryogenesis are described. Emphasis is placed on the description of the cell surface antigens of several cell lines. Syngeneic antisera raised against two primitive lines (F9 and PCC4) and against a differentiated one (Endo) have allowed the detection of three groups of cell surface antigens, present on teratoma cells, tumor cells and embryonic cells. The F9 antigen appears to be specific to be specific of the very early steps of egg development (morula and blastocyst). After egg implantation, it keeps expressed on the cells of the male germ line. The PCC4 antigen has a similar cell type distribution but appears to be more specific of multipotential cells. The Endo antigen is essentially specific of endodermal derivatives. The F9 antigen is probably specified by the wild type allele (+ tl2) of the tl2 gene at the T-Locus of the mouse, a gene which plays some critical role in early development. The molecular structure of this antigen, as determined from immunoprecipitates is very similar to that of H-2 antigens. In addition, a cross-reacting material is found in Man, with a tissue distribution identical to that found in the mouse.


Preclinical warning of recrudescent mammary cancers by pregnancy-associated alpha-macroglobulin. Nine of 30 mammary cancer patients developed metastases during 13-94 months after mastectomy. All 9 patients had elevated blood levels of pregnancy-associated alpha-macroglobulin (PAM) 1-21 months before conventional detection of metastases. Seven of the clinically well patients had PAM rises exceeding 90 per cent above the baseline and in 4 of these the PAM later fell to lower levels. PAM appears to have potential as an indicator of the growth of micrometastases.


Regional chemotherapy with combined drugs in cancer of the head and neck. Fifteen patients were treated with intra-arterial fluorouracil, methotrexate, and bleomycin in combination for palliation of advanced cancer of the head and neck. In all cases irradiation or irradiation plus surgical resection had previously failed to control the tumor. Tumor regression occured in 87% of the cases (13/15), and was complete in three (20%). Regressions lasted for up to 13 months. In four instances tumor regression permitted radical resection. The response rate achieved with this drug combination was greater than that observed earlier using each drug individually. Intra-arterial chemotherapy provides useful palliation for advanced head and neck cancer and the results can be improved with drug combinations.


Attempt at local administration of anticancer agents in the form of fat emulsion. A fat emulsion when injected into tissue is scarcely taken up by the blood vascular system but is retained within the tissue over a relatively extended period, and is distributed slowly into the surrounding tissues and to the regional lymph nodes. Attempts were made to use this property of the emulsion in the local administration of anticancer agents in emulsion, both in experimental animals and in man. The concentrations of bleomycin in the tumor tissue of rats were significantly higher after the intratumoral injection of the emulsion form than when the drug was administered in the aqueous solution, either systemically or intratumorally. Experimental antitumor activity against this tumor was superior after the bleomycin emulsion, as well. In the clinical trials six of eight patients with either squamous cell carcinoma of skin or local recurrence of adenocarcinoma of the breast responded favorably to this treatment.


Intravenous bleomycin infusion as a potential syncronizing agent in human disseminated malignancies: a preliminary report. According to cell cycle synchrony principles, bleomycin was infused for 48 hours, followed by a dose of either methotrexate or hydroxyurea after a 24-hour rest, in 36 adult patients with disseminated carcinoma. In this preliminary study, a 59% response rate was noted among patients with epidermoid carcinoma of the head and neck. Four of four patients with transitional cell carcinoma of bladder and one patient with hypernephroma also responded. No responses were noted among five patients with epidermoid carcinoma of the lung. The length of response ranged from 1 to 8 months (median, 2 months). Seventy-seven percent of the responders had extensive prior radiotherapy. The first patient treated had fatal sepsis with leukopenia, which prompted a widening of the treatment interval. Subsequently, toxicity was mainly mild or absent, the moderate or severe toxicity was primarily neutropenia, which was reversible. The use of low-dose bleomycin infusion is safe and may play a role in cancer therapy in combination with other agents specific for certain tumors. The length of infusion should be determined by the cell cycle of the tumor, if its potential synchronizing capabilities are to be exploited.


Prognostic value of alpha-fetoprotein radioimmunoassay in surgically treated patients with embryonal cell carcinoma of the testis. Alpha-fetoprotein was assayed radioimmunologically in 51 samples of sera from 26 patients who had been operated for embryonal cell carcinoma of the testis. The test was found to have good prognostic value. Elevated levels were seen frequently in patients with metastase or who developed metastases. The kinetic study of alpha-fetoprotein allows us to monitor treatment efficiency, as well as to study cancer evolution.


Immunofluorescent demonstration of alpha- fetoprotein and other plasma proteins in yolk sac tumor. All seven pure yolk sac tumors of gonadal and extragonadal origin tested showed a bright positive fluorescence for alpha-fetoprotein in the tumor tissue. A positive reaction was seen in both the tumor cells and the hyaline globules. In all cases, however, the positive fluorescence was distributed in some focal areas of the tumor tissue. Certain tumor cells showed a strong granular intracytoplasmic fluorescence, whereas others showed a weak or a negative fluorescence. The fluorescence-positive tumor cells were located mainly in the areas rich in fluorescence-positive hyaline globules. Besides alpha-fetoprotein, certain plasma proteins--albumin, alpha-1 antitrypsin, and transferrin--were also demonstrated in all five yolk sac tumors tested. The pattern of the distribution of positive fluorescence was basically similar to that of alpha-fetroprotein. Other plasma proteins--orosomucoid, haptoglobin, Gc-globulin, alpha-2 macroglobulin, hemopexin, and ceruloplasmin--were present in certain tumors, and were distributed mainly in a limited number of hyaline globules. Both IgG and IgA were present in two tumors of ovarian origin. The immunoglobulins were for the most part present in extracellular hyaline globules, suggesting that these are taken up from the circulation. Test for fibrinogen, beta-lipoprotein, IgM, IgE, beta-1C/beta-1A and beta-1E globulins were negative or questionable. In a hepatoblastoma, tests for alpha-fetoprotein were positive, but those for other plasma proteins were negative. Fine granular fluorescence was seen in each hepatocellular tumor cell. Mesenchymal elements were virtually unstained.


Intracavitary bleomycin in the management of malignant effusions. Instilled bleomycin and thoracostomy were utilized in 38 patients with malignant pleural effusions; the therapy produced a complete or partial response rate of 63%. Toxicity was minimal. In patients with intraperitoneal effusions, bleomycin instillation after drainage produced a complete or partial response in 36%. One patient had severe hypotension and fever. Patients with ovarian and breast carcinoma responded best, among them, effusions were controlled in greater that 70%. Because of its low systemic toxicity, absence of marrow toxicity, and virtual absence of discomfort, we think that the local instillation of bleomycin is indicated in the management of malignant effusions.


Choriocarcinoma: expression of tumor- and trophoblast-associated antigens in patients with low chorionic gonadotropin excretion. The circulating levels of four tumor- or trophoblast-associated antigens were measured by specific radioimmunoassays in 11 patients with gestational choriocarcinoma. The estimations were carried out at the time when the urinary gonadotropin (hCG) excretion was low or negligible. Gonadotropin, measured as the hCG beta-subunit, was detected in serum of three patients, one of whom also showed a slightly raised level of carcinoembryonic antigen (CEA). All patients had normal serum alpha-fetoprotein (AFP) levels and no trace of human placental lactogen could be demonstrated. Repeat estimation after treatment of patients with raised levels showed a disappearance or a marked decrease of the circulating hCG levels and a return to normal of the elevated serum CEA level. The results show that although CEA levels may occasionally be elevated new information can hardly be expected from markers other than hCG when one is monitoring response to treatment, but AFP may have potential significance in the distinction between pregnancy and a trophoblastic disease. The circulating levels of hCG are of vital importance in the monitoring of choriocarcinoma patients who appear to be in remission by the conventional analysis of urinary hCG excretion.


Chemotherapy versus chemoimmunotherapy of head and neck cancer: report of a randomized study. Thirty-four patients with recurrent squamous cell carcinoma of the head and neck were randomized to receive the five-drug chemotherapy regimen BACON (bleomycin, adriamycin, CCNU, vincristine [Oncovin], and mechlorethamine [nitrogen mustard]; 14 patients) or the same regimen plus bacillus Calmette-Gu erin (BCG) by scarification (20 patients). The majority of both patient groups had received prior surgery and radiation. The patients treated with BACON plus BCG experienced a significantly longer survival (P = 0.014) than those treated with BACON alone. There were five drug-related deaths, and eight other patients required hospitalization for treatment of drug-related morbidity.


Lower plants as a source of anticancer drugs. The NCI's antineoplastic fermentation program is reviewed. Clinically useful antitumor antibiotics are discussed in terms of their activity, mechanism of action, and the producing microbe.


Osmium zinc iodide staining of Golgi elements in oocytes of Triturus cristatus. Developing oocytes of the newt Triturus cristatus were studied in order to clarify the role played by the Golgi apparatus in the formation of yolk. The cytochemical method used for this purpose was that of Maillet (1968) which employs an Osmium Zinc Iodide (OZI) complex. Previtellogenic oocytes reveal a pattern of OZI staining only after hormonal (HCG) stimulation, following which both the Golgi apparatus and the multivesicular bodies are stained. Vitellogenic oocytes taken from non-hormonally stimulated females reveal OZI deposits in a number of vesicles peripheral to the Golgi apparatus as well as within the superficial layer of the forming yolk platelets. Following hormone stimulation, many of the Golgi apparatus located in the central ooplasm of vitellogenic oocytes have all their cisternae blackened by the OZI deposits; other apparatuses,more peripherally located, remain essentially unchanged in their staining pattern. Further, a large number of OZI stained vesicles becomes visible in the vicinity of the Golgi apparatus and within the superficial layer of the forming yolk platelets. The present findings are interpreted as indicating the occurrence of fusion between Golgi derived vesicles and forming yolk platelets. It is also suggested that the vesicles in question function as carriers of Golgi produced enzymes which are presumably required to accomplish the final elaboration of the yolk material.


The desmosome: fine structural studies with freeze-fracture replication and tannic acid staining of sectioned epidermis. Desmosomes of larval and post-metamorphic newt epidermis have been studied by freeze-fracture replication both with and without prior glutaraldehyde fixation. Characteristic particles of a diameter (70-130 A) similar to that of typical membrane associated particles are found clustered on the exposed internal faces of adherent desmosomal membranes. They remain attached to the B-face in unfixed material, but occupy the desmosomal A-face after fixation. Membrane associated particles of nondesmosomal surfaces are found predominantly on the A-face in both fixed and unfixed epidermis. Suitably oriented replicas of unfixed desmosomes reveal profiles of apparent fine filaments extending from the region of tonofilament loops through the desmosomal plaque to traverse the cytoplasmic leaflet of the plasmalemma. They can be traced onto the B-face. Their position correlates to fine linear profiles noted in tannic acid/glutaraldehyde-fixed and sectioned desmosomes. The possibility that these represent a mechanism for anchorage of tonofilaments to the plaque and to the membrane is discussed. These and other fine structural features are compared and contrasted to the properties of hemidesmosomes described in the preceding report.


The Kurloff cell. Its differentiation in the blood and lymphatic system. Kurloff cells belong to the group of macrophages as far as ultrastructure, adhesiveness and identification with Kupffer cells (in the case of the liver) are concerned. A characteristic group of features makes it easy to recognize them: PAS reaction, cyanol-blue staining, presence of myelin figures in electron microscopy and attachment to glass-slides. Kurloff cells are thymic and blood cells. They are observed in small numbers in the circulating blood and, in large quantities, in spleen (red pulp), liver (hepatic sinusoids) and lung (septal capillaries). They are absent from lymphatic nodules and from diffuse lymphatic tissues. Morphological and experimental studies indicate that, in spite of some equivocal similarities, Kurloff cells are distinguishable by many criteria from erythrophagocytic cells and from protein-secreting blood cells.


The zinc iodide-osmium tetroxide (ZIO) reaction on nerve endings in the median eminence of the rat under normal and experimental conditions. The reaction of nerve endings in the median eminence of the rat to zinc iodide-osmium tetroxide (ZIO) staining was examined electron microscopically under normal and experimental conditions. The experimental condition of catecholamine exhaustion in the nerve endings was induced by the administration of H44/68 and reserpine. Vesicles in the terminals of catecholaminergic nerves reacted similarly to ZIO staining in both normal and experimental material. The majority of synaptic vesicles in various terminals gave a positive ZIO reaction. The neurosecretory elementary granules, however, failed to react with ZIO. On the other hand, some nerve terminals in the external layer of the median eminence showed a strong positive reaction in the cytoplasmic matrix, in mitochondria as well as in synaptic vesicles. These findings strongly suggest that the ZIO-positive substance in nerve terminals is not the transmitter itself, i.e. the monoamine, but rather represents a range of substances commonly found in various kinds of synaptic vesicles and is probably proteinaceous in nature. A brief discussion is also given on the difference in ZIO reactivity between neurosecretory elementary granules and small vesicles in the hypothalamo-hypophyseal tract.


Endogenous type C RNA virus of Odocoileus hemionus, a mammalian species of New World origin. Type C RNA viruses have been isolated from several Old World vertebrates, and an even larger number of Old World species have been shown to contain endogenous viral genetic sequences. The present report describes the first isolation of type C virus endogenous to a species originating in the New World. This virus, isolated from cells of the Columbian black-tailed deer, Odocoileus hemionus, is shown to possess genetic sequences in common with DNA of its species of origin. While it shares biochemical and immunologic characteristics with other mammalian type C viruses, its immunologic properties readily distinguish it from known endogenous viruses.


Antigenic variants of influenza A virus obtained in vitro. The aim of this study was to investigate antigenic "drift" in the haemagglutinin and neuraminidase antigens of influenza A virus in vitro under immunological pressure. Variants of the "Asian" influenza strains A/England/12/64 (H2N2) and A/Tokyo/3/67 (H2N2) were isolated in the allantois-on-shell system in the presence of homologous postinfection ferret sera. For each of these two viruses three generations of variants were isolated and characterized. It was found that the successive antigenic variants of A/Eng/12/64 did not resemble A/Tokyo/3/67. Thus it is probable that the pathway of antigenic drift in vitro was not the same as that which occurred in nature during the evolution of A/Tokyo/3/67 from A/Eng/12/64. In addition, A/Tokyo/3/67, which was the last strain to be prevalent before the A/Hong Kong subtype appeared, underwent significant antigenic drift from "junior" to "senior" variants. This finding did not support the concept that, when antigenic drift occurs, resulting in the appearence of viruses with new haemagglutinin antigen subtypes, the previously prevalent strain has no capacity for further antigenic drift. The study did not result in the production of strains that were identifiable as "bridging" viruses between the H2 and H3 haemagglutinin subtypes. The present paper includes the first report of antigenic variation in the neuraminidase antigens of influenza A viruses occurring in vitro under immunological pressure.


[Cyto-immunologic location of alpha-MSH and ACTH in the lead-hematoxylin (HPb)-stainable pituitary cells, in the eel]. The lead-haematoxylin positive cells of the pars intermedia react with anti-alpha-MSH and anti-1-24ACTH or anti-17-39ATCH; those of the rostal pars distalis are only revealed with antisera anti-1-24ACTH and anti-17-39ACTH. Intensity of cytoimmunological staining, which is modified after various experimental treatments (reserpine, metyrapon, pimozid, cortisol...) and during black or white background adaptaiton, corresponds essentially to that of the PbH staining.


[Study of the nephrotoxicity of antibodies directed against the glomerular basement membrane glycoproteins of the Wistar rat]. The nephrotoxic activity of prepared antibodies against previously isolated glycoproteic fractions of the Rat glomerular basement membrane was studied using Masugi's model of Nephritis. This activity was determined by following the daily evolution of the proteinuria and the value of the seric complement. It seems linked to numerous factors.


[Immunosuppressive effect of a human serum ferroprotein of hepatic origin: alpha-2-HF-globulin]. alpha 2HF, a serum  ferroprotein increased during some malignant diseases, displays a suppressive effect on in vitro antibody responses, whether T-independent or T-dependent. This effect is not related to the presence of iron, nor to the macroglobulin nature of the molecule. The immunosuppressive properties of alpha 2 HF might depend on its carbohydrate moeity.


[Levels of lead on the livers of dogs from the Paris region]. Lead level has been determined in the liver of 51 Dogs of different ages, sex and breed from districts of Paris. This level is from 0,16 ppm (fresh weight) up to 5,4 ppm. 23 p. cent of Dogs examined have more than 3 ppm in their livers. The level is not correlated with sex, breed and size. It increases with age of animals.


[Determination by three technics of cellular immunology of the antigenic determinants of the Ly-Li system expressed on human B lymphocytes]. A clear correlation has been observed between the presence of the antigenic B cell system Ly-Li detected serologically, and 3 cellular immunology techniques: 1. MLR inhibition by anti-Li serum; 2. level of restimulation of anti-Ly-Li in vitro primed lymphocytes; 3. detection of HLA-D alleles by homozygous typing cells. These results suggest that the allelic products detected serologically may be identical to those detected by the first two techniques, namely MLR inhibition and in vitro primed lymphocyte typing, and possibly HLA-D typing using homozygous typing cells, although the correlation was found to be repeatedly less clear for the last technique.


Studies on J chain and binding site for secretory component in circulating human B cells. Lymphocyte-enriched mononuclear cell fractions were obtained from the peripheral blood of four healthy young adults. Living B cells with membrane immunoglobulin (Ig) were studied by immunofluorescence to reveal exposed J-chain determinants or surface affinity for the secretory component (SC). Alcohol-fixed cell smears were similarly studied with and without prior denaturation in acid urea. It was concluded that J chain-containing polymeric Ig of endogenous origin is generally not present on the surface of circulating B cells. If occasional cells bear IgM or IgA polymers rather than monomers, their SC-binding site must be concealed to a degree that it is not functional. Affinity for SC is therefore unlikely to be involved in a selective homing of IgM- and IgA-immunocyte precursors from peripheral blood to glandular regions.


Studies on J chain and binding site for secretory component in circulating human B cells. II. The cytoplasm. About 0-3-1-1% of the lymphoid cells from peripheral blood of healthy adults contained cytoplasmic immunoglobulin (Ig). The class distribution of these B cells varied greatly among individuals, with a preponderance of the IgA (26-65%) or the IgG (15-66%) class. A remarkably high percentage of the Ig-containing cells were positive for cytoplasmic J chain regardless of the class (100% for IgM, 87-97% for IgA, 50-100% FOR IgD, and 43-88% for IgG cells). This feature probably reflects that the cells represent circulating blasts derived from the early expansion phase of B-cell clones. The antigenic determinants of the J chain were in most IgA-containing cells considerably masked, indicating that this subunit was "correctly" arranged in the IgA dimers at the cytoplasmic level in the manner demonstrated for intestinal IgA plasma cells.


Restriction of immunoglobulin heterogeneity, autoimmunity and serum protein levels in aged people. Ninety-one sera of persons above 80 years of age were screened for autoantibody activity against lipoproteins (anti-LDL 7, anti-HDL 6 positive), for rheumatoid factor activity (Latex 14, Waaler-Rose 7 positive) and for antinuclear factors (11 positive). Among the sera with autoantibody activity 29 percent showed deviations of the normal kappa/lambda ratio of immunoglobulins, as opposed to 22 percent of the sera without detected autoantibody activity. In 3 percent of the sera an M component was detected. Determination of the alpha1-acid glycoprotein, alpha1-antitrypsin, haptoglobin, haemopexin, complement component C3c and C4, IgG, IgA and IgM levels showed significant increases in alpha-, and beta-globulins as well as in IgG and IgA in sera of the aged persons as compared to a normal population between 20 and 60 years old. No significant difference was noted between the gamma-globulin concentration in sera of aged persons with or without autoantibody activity. The evaluation of the relationship between serum protein levels and alterations of the kappa/lambda ratio indicated that the alpha- and the beta-globulins were significantly raised in sera with altered kappa/lambda ratios, whereas, with the exception of M component containing sera the gamma-globulin levels seemed not significantly affected by changes in this ratio.


Effects of Hoechst 33258 on human leukocytes in vitro. The benzimidazole derivative Hoechst 33258 was added at various concentrations to human leukocyte cultures. After 16 or 24 h of treatment, with concentrations equal to or greater than 100 mug/ml of Hoechst 33258, a number of chromosomes showed regions in which the chromatin was undercontracted. The centromeric regions of chromosome 1 and, more rarely, of chromosomes 3 and 9 appeared to be decondensed. Short decondensed regions were also present on the long arms of chromosomes 1 and 2. The possible nature of these regions is discussed.


[Total gastrectomy in patients over 70 years old. Postoperative complications and prognosis]. In the cases of gastric cancer in patients over 70 years old we were not able to find a noticeable difference of survival time between the cases of palliative total gastrectomy and those where no resection or no operation was undertaken. From these results and the high frequency of complications after total gastrectomy in patients over 70 years old, we cannot see the therapeutic significance of palliative total gastrectomy. On the other hand the results of postoperative examinations in the cases without cancer recurrence were favorable, and they live a relatively comfortable life for their age. If the preoperative risk of patients can be judged exactly and if cure can be expected, total gastrectomy must be considered even in the patients over 70 years old.


[Clinical aspects of Hochenegg's draft procedure]. A report concerning the follow-up examinations of the 14 patients with rectum cancer from the University Surgical Clinic in Innsbruck, Austria, who from 1961-1974 were operated on using Hochenegg's technique. Interestingly enough, it was noticed that when the height of the anastomosis was less than 5 cm from the anal ring, a partial incontinence appeared (in 3 of the 14 patients) and when it was more than 5 cm from the anal ring, a total continence was assured. A relapse was observed three times in the tumor cases type C2 and D (Duke). Post-operative complications occurred in 4 cases, none being life-threatening. The indications of this technique will be discussed.


The agglutinating antibody response in the duodenum of infants with enteropathogenic Escherichia coli gastroenteritis. The agglutinating antibody response in duodenal fluid and serum was measured serially in 15 infants with enteropathogenic Escherichia coli gastroenteritis. Peak levels of duodenal agglutinins were recorded 8-18 days after the onset of symptoms, and the titres fell within the next 7-14 days. The immunoglobulin (Ig) class of these antibodies was mainly IgA, but IgM antibodies were detected early in the response, especially in the younger infants. Late antibodies showed less cross-reactions with other strains of E. coli than did early antibodies. Serum antibody responses were detected in eight infants, but they correlated poorly with the titres of intestinal antibodies. No rise in serum antibodies was detected in six infants. It is not known whether these differences are host-derived, or whether they are the result of the invasive properties of some of the infecting organisms.


Studies on the pathogenesis of enteric infections caused by invasive bacteria. Salmonellae, shigellae and some Escherichia coli must invade the intestinal epithelial cell and multiply within the mucosa to cause disease. Although the bacterial cell most likely possesses several properties essential to this invasive ability, the nature of the cell envelope complex is at present the only characteristic which has been implicated in this process. While a number of pathophysiological events result from invasion, some of our recent efforts have concerned the site and mechanism of intestinal fluid loss in salmonellosis and shigellosis. In both these disorders, bacterial invasion of the colonic mucosa, associated with an acute inflammatory reaction and mucosal damage, is regularly seen and colonic salt and water transport is abnormal. These defects may account for mild diarrhoea in salmonellosis and the dysenteric stools of shigellosis. However, in salmonella-infected animals with severe watery diarrhoea and in shigella-infected animals with diarrhoea alone or in combination with dysentery, the jejunum is in a net secretory state. This secretion occurs in the absence of bacterial invasion or morphological abnormalities. Thus, the diarrhoea caused by invasive bacteria may result from the inability of the colon to reabsorb the increased volume of fluid entering it from the small intestine. Although colonic mucosal damage is a feature of invasive-type diarrhoeas, the permeability of both the colon and small intestine to small molecules, mannitol and erythritol, is not altered. Thus intestinal fluid loss cannot be ascribed to transudation. In addition, the results of our Ussing chamber experiments, employing salmonella-infected rabbit ileum, reveal that salt and water secretion is an active process. Since secretion occurs in the jejunum in the absence of bacterial invasion, this might suggest the participation of an enterotoxin. Shigella dysenteriae I is the best-studied invasive organism in which an enterotoxin has been found, yet mutant strains which do not invade but retain the ability to elaborate enterotoxin fail to cause disease in either monkeys or man. Thus, the physiological relevance of Shiga enterotoxin and the mechanism of jejunal secretion in these disorders remain unclear. Recent data suggest that invasive enteropathogens, like the enterotoxin-producing bacteria, activate the mucosal adenylate cyclase-cyclic AMP system and that this activation may play a role in intestinal fluid secretion.


[Results of operative treatment of bronchial carcinoma (author's transl)]. Between 1960 and 1974 a total of 14 937 patients were treated in 16 German surgical departments for bronchial carcinoma. 70% proved to be inoperable. In 30% the tumour was resected, the five-year survival rate of those patients operated on before 1969 being 23%. In suitable cases, lobectomy proved to be a satisfactory radical method of resection in parallel with pneumonectomy, while the operative risk of the latter was twice as high. The operative risk decreased by half during the period of observation. Best results of tumour resection were achieved when in the early stage. The prognosis of anaplastic carcinoma was not significantly worse than that of other histological types. Further improvement in the results of surgical treatment of bronchial carcinoma are to be expected if the operative risk can be decreased further and more patients can be operated on at an earlier stage.


[Results of radiotherapy of bronchial carcinoma (author's transl)]. In a joint retrospective study by 17 radiotherapy clinics in German-speaking countries the results of treatment of bronchial carcinoma after radiotherapy were analysed in 7503 cases. The age peak was between the 60th and 70th year. Squamous-cell carcinoma was the most frequent histological type, followed by anaplastic carcinoma, with adenocarcinoma being rare. There was a high proportion of histologically not clearly identified cases (27% in central and 35% in peripheral carcinomas). Survival rate at one year was 31% for central (3662 patients) and peripheral (961 patients) tumours, but only 2% at five years. Prognostically there was no difference between histological types and kind of radiotherapy or technique, but total dose affected survival rate. At a total dose of less than 5000 rd the survival rate at five years was minimal. The prognosis of combined surgical and radiotherapeutic measures was slightly better than with a radiotherapy alone, but results were unpredictable for the individual case. It is concluded that radiotherapy aiming at cure should be used in imoperable bronchial carcinoma if the tumour state and general condition of the patient appear to make a cure possible. But if this is not the case, radiotherapy should be used only palliatively, i.e. only to ameliorate symptoms.


The role of endogenous catecholamines in the regulation of electrocortical activity in the encephale isole cat. Experiments were performed on encephale isole cats. The cats were either untreated or pretreated with reserpine, alpha-methyl-p-tyrosine (AMPT) or FLA 63, and the depletion of endogenous noradrenaline was determined by fluorescence microscopy. Pretreatment with reserpine reduced waking and spindle sleep while the synthesis inhibitors did not. A combined pretreatment with reserpine and AMPT or FLA 63 was necessary to deplete totally brain stem noradrenaline. Under these circumstances, behavioural arousal was abolished and all electrocortical activity except spindling was also abolished. Injection of L-DOPA into cats pretreated with reserpine and AMPT produced behavioural alerting and low voltage, high frequency electrocortical activity. In cats pretreated with reserpine and FLA 63, the minimum effective dose of L-DOPA was often elevated, and when the low voltage, high frequency activity occurred, it was accompanied by spindling. It was accompanied by spindling. It is suggested that dopamine and noradrenaline independently modulate electrocortical activity, dopamine mediating spindling, and noradrenaline inhibiting spindling but mediating low voltage, high frequency activity.


Influence of association cortex on penicillin discharges in the primary visual cortex. The influence of visual association cortex on the pattern of neuronal activity in the lateral geniculate body, during penicillin-induced discharges in visula cortex, was studied. In a majority of those geniculate cells which exhibited a pre-penicillin discharge increment in activity, that increment was abolished or significantly reduced during focal hypothermia of the visual association cortex. The frequency of penicillin discharges in visual cortex was, in most cases, reduced during cooling of association cortex. These findings are briefly discussed in terms of corticothalamic interactions.


Epilepsy and sleep organization in the baboon Papio papio. Effects of different types of epileptic seizure on the sleep organization of baboon were studied through 12 h polygraphic recordings. Photically induced myoclonic jerking did not modify sleep organization. Photically induced genalized convulsions produced an increase of the waking stage (AWA) and reduction of REM stage. Pentetrazol-induced generalized convulsions not only increased AWA and reduced REM, but also elicited various modifications of stages I, II and III. Daily generalized convulsions induced by light or pentetrazol over 5 days demonstrated no rebound of REM.


Automated sleep EEG analysis applied to the evaluation of drugs: illustration by study of clorazepate dipotassium. An automated sleep EEG analysis system was used to evaluate the effects of clorazepate dipotassium in normal subjects. Ten young-adult men slept 18 consecutive nights in the laboratory. On days 8-15 clorazepate (7.5 mg) was administered three times daily; on days 5-7 and 16-18 a placebo was administered in a similar fashion. The drug reduced amounts of alpha and delta activity and increased the amount of beta activity and the number of spindles. These effects generally persisted through the 3 day placebo recovery period. Our results suggest that sleep EEG waveform descriptors are sensitive indicators of drug activity and that beta activity in particular may be useful in the detailed description of various drug effects.


Correlation between confirmed sites of neurological lesions and abnormalities of far-field auditory brainstem responses. Far-field auditory brainstem responses were recorded in ten patients in whom the distribution of pathology was defined at autopsy or at operation. The response normally consists of seven components in the initial 10 msec following click signals. Interruption of audiotory pathway at the junction of VII nerve with brainstem results in loss of response components after Wave I. Interruption of auditory pathway at the midbrain results in loss of response components after Wave III. We conclude that Wave I reflects activity of VIII nerve, Waves II and III reflect activity of cochlear nucleus, trapezoid body, and superior olive and Waves IV and V reflect activity of lateral lemniscus and inferior colliculus. The generators of Waves VI and VII were not defined.


Spatial distribution of potentials evoked by half-field pattern-reversal and pattern-onset stimuli. Spatial distributions of visual potentials (VEPs) evoked by half-field checkerboard pattern-reversal and pattern-onset stimuli were studied in 13 subjects, using an 11 lead unipolar array. The main aim was to confirm findings, obtained by previous workers with bipolar recordings, that half-field pattern-reversal VEP's are confined to the contralateral hemisphere and that half-field pattern-onset VEPs are asymmetrical, with greater right hemisphere involvement. Pattern-reversal VEP's contained four consistent peaks, designated here by polarity and peak latency as: (a) P95, positivity contra-lateral and negativity ipsilateral to the field stimulated; (b) P125, predominantly ipsilateral positivity; (c) N165, predominantly ipsilateral negativity; (d) P225; predominantly midline positivity. Pattern-onset VEPs contained three consistent peaks: (a) P125, mainly contraleteral positivity; (B) N175, mainly contralateral negativity; (c) P225, midline positivity. Distributions of pattern-reversal and pattern-onset peaks resembled one another only for P225, suggesting different cortical representation for the other events of the two kinds of VEP. Bipolar pattern-reversal VEPs were largely contralateral, but unipolar recordings showed that this was due to steeper contralateral potential gradients, as ipislateral activity was widespread. Pattern-onset peaks did not differ in amplitude with respect to the half-field stimulated. Previously reported asymmetries were not confirmed. The P125 and N175 pattern-onset peaks were almost entirely restricted to the contralateral hemisphere, but the distributions by half-field were mirror-images of one another. Half-field pattern-onset stimuli could be used to investigate the responsiveness of each hemisphere, although differential hemispheric involvement was not shown. Several differences in amplitude and distribution resulted from varying the width of the vertical central dark strip.


Temporo-spatial propagation of epileptoform after-discharges in the isolated cat suprasylvian gyrus. 1. Epileptiform after-discharges (EADSs) induced by electrical stimulation of the isolated suprasylvian gyrus were studied in cats with chronically implanted electrodes. 2. In a given region and at a certain time after stimulation, the following events took place: (a) a slow radial spread of the zone of maximal depolarization, from the cortical surface downward, as evidenced by a laminar study; (b) a massive cellular discharge preceded by a period during which few unit activities were detected, followed by bursts of spike activity timed with the surface-positive waves of the ECoG; (c) a surface-negative DC shift with maximal amplitude around 1000 mum below the surface; (d) the occurrence of a synchronizing focus from which the paraoxysmal waves propagated to the whole gyrus. 3. All these phenomena spread across the surface of the gyrus with a velocity (7-20 mm/min) similar to that of focal seizures in man.


Cortically elicited spike-wave after discharges in thalamic neurons. Self-sustained, 3-4/sec spike-wave ADs were elicited in lateralis intermedius-lateralis posterior thalamic bursting neurons following incremental responses elicited by 10/sec shock-trains applied to the anterior suprasylvian cortex. The pattern of cortically elicited thalamic spike-wave complexes, with brief depolarizing components and a long-lasting hyperpolarizing wave, resembles that of previously described spike-wave seizures elicited in cortical interneurons following specific thalamo-cortical augmenting responses.


The immediate uptake of potassium ion by mitochondria requiring gramicidin and 2,2-dimethylsuccinic anhydride. The addition of 2,2-dimethylsuccinic anhydride to mitochondrial suspensions fortified with gramicidin and potassium ion but without any permeant anion caused an immediate and rapid increase in volume (as indicated by absorbance change at 520 nm) and the uptake of potassium ion (as indicated by a cation-specific electrode). The phenomena was not inhibited by rutamycin but was inhibited by either rotenone, antimycin or 2,4-dinitrophenol. Rotenone inhibition was relieved by succinate thus one of the requirements of the process was energy derived from endogenous substrates. Potassium ion could be replaced by rubidium and cesium ions but not by lithium or sodium ions. Since 2,2-dimethylsuccinate could not replace the anhydride and was not a permeant anion there must also be a requirement for the anhydride bond. The action of the anhydride on the mitochondria must be direct. Only closely related anhydrides were capable of engendering the effect of a highly effective permeant anion.


Structural features of encephalomyocarditis virus RNA from analysis of reverse transcription products. The presence in encephalomyocarditis (EMC) virus RNA of homonucleotide tracts 10 nucleotides or more in length has been investigated by testing the ability of homo-oligodeoxynucleotides to prime DNA synthesis in the reverse transcriptase from avian myeloblastosis virus. Neither (dC)10 nor (dA)10 promoted incorporation of [3H]deoxynucleotides into acid-insoluble material but (dG)10 and (dT)12-18 were effective primers and produced DNA products approximately 2000 nucleotides in length. We conclude that there are single-stranded oligo(rC) and oligo(rA) tracts in native EMC virus RNA at 37 degrees C. Kinetic analysis indicated that oligo(dT) priming is similar to priming on ovalbumin mRNA and that it gives rise to only one DNA product per template molecule. Oligo(dG) priming appears to be complicated by self-aggregation of the primer. Oligo(dT)-primed and oligo(dG)-primed DNA have both been separated on alkaline-sucrose gradients into two peaks of which only the 'heavier' will hybridise to EMC virus RNA. Competitive hybridisation experiments indicate that the 'heavy' oligo(dT)-primed and oligo(dG)-primed DNA fractions hybridise to overlapping sequences of EMC virus RNA and place the priming regions of EMC virus RNA approximately 500 nucleotides apart during reverse transcription.


Anaphylactoid reactions due to hydroxyethyl starch infusion. Incompatibility reactions due to hydroxyethyl starch (HES) were observed during 8 out of 10,273 infusions of 500 ml 6% HES (Plasmasteril). The clinical symptoms ranged from skin reactions to tachycardia, hypotension and shock. In 3 of the 8 patients with incompatibility serum immunoglobulin concentrations were reduced after the anaphylactoid reaction. Specific antibodies against HES were, however, not detected. Serum IgE levels stayed within their normal limits. Positive reactions of the immediate type to intradermal skin tests with different dilutions of Plasmasteril were obtained in five patients.


H-2 antigen-specific cytotoxic T cells induced by concanavalin A: estimation of their relative frequency. Specific and nonspecific lysis of DBA/2 (H-2d) mastocytoma cells, P815, by concanavalin A (Con A)-induced cytotoxic T cells was studied. In the assay for nonspecific lysis, phytohemagglutinin (PHA) was present to glue the target and killer cells together. We have presented evidence previously to show that PHA reveals only, and all, cytotoxic T cells. In the assay for specific lysis the only glue present was specific receptors on a fraction of the killer cells and surface antigens of P815. We show that when PHA was present, Con A-induced cells which were syngeneic, semi-syngeneic, or allogeneic, lysed P815 very efficiently in a 4-h 51Cr release assay. However, only Con A-induced T cells which were allogeneic and did not carry H-2d lysed P815 when the assay was carried out in the absence of PHA. In an experiment with two target cells, Con A-induced B10 (H-2b) T cells lysed B10.D2 (H-2d) targets specifically but did not lyse B10 targets, while Con A-induced B10.D2 T cells lysed B10 targets specifically but not B10.D2 targets. Furthermore, Con A-induced B6 (H-2b) T cells from normal mice lysed P815 specifically but Con A-induced B6 T cells from irradiated F1 (B6 x BALB/c) (H-2b/d) mice reconstituted with B6 bone marrow did not lyse P815 specifically. A fraction of Con A-induced T cells therefore appear to bear specific surface receptors for nonself H-2 coded structures. We describe conditions of assay and a new method of plotting the results such that nonspecific (PHA-revealed) and specific (PHA-independent) cytotoxicity can be quantitatively compared. We conclude that 1-4% of the total Con A-induced cytotoxic effector T cells are directed against any particular foreign H-2 haplotype. This is the first estimate of the relative frequency of antigen-reactive cytotoxic T cells.


Isolation of antigen-binding cells from unprimed mice. II. Evidence for monospecificity of antigen-binding cells. Spleen cells from unimmunized mice were exposed to two contrastingly fluorescent antigens simultaneously. Antigen-binding cells of either specificity were isolated using a fluorescence-activated cell sorter (FACS). Purified cells binding one or the other of the antigens were then examined by fluorescence microscopy for the presence of bound antigen of the alternate specificity. No double binding cells were seen. If cells bear receptors of two or more specificities and these receptors are randomly distributed among antigen-binding cells, then of the 13 000 binding cells examined 82 were expected to bind both antigens. These results strongly suggest that antigen-binding cells bear receptors of only one specificity. In addition, by inference from the functional correlation between antigen-binding cells and precursor cells, the data support the contention that precursors of antibody-forming cells are monospecific.


Suppressor cell induction in vitro. I. Kinetics of induction of antigen-specific suppressor cells. The induction of antigen-specific suppressor cells in vitro, using high concentrations (100 mug/ml) of keyhole limpet hemocyanin (KLH) in Marbrook flasks is described. Spleen and cortisone-resistant thymocytes were the richest source of suppressor cell precursors, compared to lymph node cells, peripheral blood lymphocytes or thoracic duct lymphocytes. Suppressor cells induced with KLH only suppressed KLH-reactive helper cells, and not B cells or helper cells of other specificity. The suppressor cells were T cells, as judged by their sensitivity to anti-Thy-1.2, heterologous anti-T, but not anti-B antisera.


Solubilization and molecular characterization of membrane-bound antigens shared by thymocytes and brain. In order to characterize chemically the serologically well-defined antigens shared by thymocytes and nervous tissue of mice and rats, different solubilization procedures have been tested. Treatment of thymocyte membranes with urea-Nonidet-P40 (NP40) proved to be the most valuable method, since nearly 100% of the antigen was solubilized essentially without loss of the antigenicity. By gel filtration in urea-NP40 the molecular weight was estimated to be 35 000 and the Stokes radius in sodium deoxycholate 31.8 A. In gel filtration studies a homogenous peak was obtained, whereas isoelectric focusing yielded two peaks (isoelectric points pH 4.5-5.0 and pH 5.4). The three determinants of this antigenic system were inseparable in gel filtration and isoelectric focusing, supporting the view put forward on the basis of serological studies that they are part of the same molecule. Extraction of murine thymocyte membranes with organic solvents yielded a considerable loss of activity of the thymocyte-brain antigen. There was some residual activity in the protein fraction, but none in the organic phase. Correspondingly, the highly purified ganglioside G Gtet 2b proved unable to absorb any rabbit anti-CBA brain serum. The reduced antigenic activity of the delipidated protein fraction could be restored by addition of lipids, as well as the nonionic detergent NP40. These observations suggest that interaction of protein and lipid is necessary for the antigenicity of the thymocyte-brain antigen.


Detection of immunologically active zones in equine growth hormone. Peptide fragments, obtained from equine growth hormone by cyanogen bromide cleavage and further chemical treatment, were isolated and identified. Their immunological reactivities were tested by hemagglutination and complement fixation methods using rabbit antisera against native hormone. Antigenic determinants were detected in the fragments comprising amino acid sequences 5-72 and 73-123, this last one being predominant. Fragment 124-178 had very low reactivity. Nitration of peptide 73-123 did not modify its immunological properties,but oxidation diminished them. Comparison of the antigenicity of equine growth hormone fragment 73-123 with that of the homologous ovine growth hormone fragment 86-123 lent support to the hypothesis that at least one antigenic determinant area in the horse hormone fragment is dependent on sequence 86-123.


Expression of new idiotypes following neonatal idiotypic suppression of a dominant clone. The anti-PC antibodies of BALB/c origin bear predominantly the idiotype characteristic of the phosphorylcholine (PC)-binding T15 idiotype than sera from adult mice, and, unlike the latter, they also contain detectable amounts of anti-T15 antibodies. By 2 weeks of age the anti-T15 antibodies are no longer detectable while the T15 idiotype has reached adult levels. Injection of neonatal mice with anti-idiotypic antibodies renders them unresponsive to PC until the 15th week of life. Furthermore, this treatment induces a chronic suppression of the T15 idiotype, since on recovery from unresponsiveness, the neonatally suppressed mice produce anti-PC antibodies which are predominantly T15-negative. In contrast, treatment of adult mice with anti-idiotypic antibodies induces only a transient state of unresponsiveness to PC, and the antibodies produced upon recovery bear the T15 idiotype. These findings are discussed in the context of idiotype anti-idiotype interactions and their possible role in immuno-regulation.


Many cells in rat bone marrow have cell-surface Thy-1 antigen. Thy-1.1 and Thy-1 xenoantigenic determinants were detected at the cell surface of many rat bone marrow cells. The absorptive capacity of bone marrow cells was 6-10% of that of thymocytes for Thy-1 antigenic determinants, and 30-45% of rat bone marrow cells were specifically labeled with anti-Thy-1 antibody as detected by autoradiography. Thus, while mice and rats are similar in having large amounts of Thy-1 in brain and thymocytes, they differ in that the rat lacks the antigen in most peripheral T cells and expresses it in a large number of bone marrow cells; the opposite is true in the mouse.


Tricyclic antidepressant drugs as antagonists of muscarinic receptors in sympathetic ganglia. Close arterial injection of McN-A-343 into the superior cervical ganglion of the cat resulted in contractions of the nictitating membrane. The ganglionic effects of McN-A-343 but not those of DMPP were antagonized in a dose-related manner by 2-10 mug of desipramine, imipramine, chlorimpramine, iprindole and viloxazine. No correlation was found between the dose of each drug which blocked the effects of McN-A-343 and that required to potentiate the responses of the nictitating membrane to intra-arterial administration of noradrenaline. It is concluded that clinically effective antidepressant agents can block muscarinic receptors in neural tissue, even if they do not do so in smooth muscle and gland cells.


Morphological evidence for axonal transport of glycogen in neurons innervating cutaneous receptors in Lacerta sicula (Squamata: Reptilia). Three types of glycogen-containing cutaneous nerve terminals - two of them hitherto unknown - are described. It is shown that the glycogen is synthesized in the perikaryon and transferred to the terminals by means of slow axoplasmic transport.


Comparison of a natural heparinoid with sodium and calcium heparin for their effect on the inhibitor of activated factor X. The reaction between activated factor X (Xa) and its natural inhibitor (XaI) was accelerated in vitro by both sodium heparin and an heparinoid, which was about 3 times less potent than heparin. The s. c. administration in humans of 5,000 units of sodium and calcium heparin was followed by the detection of a plasma activity potentiating XaI. In the majority of subjects, the heparinoid was not effective. These observations indicate that the use of heparinoids should not be considered as an alternative to heparin in the prevention of thromboembolism.


Study of H-2 antigens of wild mice (Mus musculus L.). 226 wild mice originating from 16 different localities were tested serologically, using a battery of anti-H-2 sera. The results indicate the existence of a large number of hitherto unknown H-2 haplotypes. Three cogenic resistant strains (B10.W44, B10.W67, B10.W625) were prepared, their H-2 haplotypes being derived from wild mice. The H-2 haplotypes of these strains are different from the haplotypes of the existing inbred mouse strains but they share with them some public H-2 antigenic specificities. An analysis of the antisera prepared against H-2wild haplotypes allowed the detection of the private antigens, H-2.107 (B10.W44), H-2.108 (B10.W67), and H-2.109 (B10. W625). Furthermore, new public antigenic specificities, H-2.60, 61, 62, were defined. All the three H-2wild haplotypes possess the Ss-high allele. Blood cells from many wild mice as well as from the individuals of the new CR strains gave positive reactions with some antisera containing antibodies against the private specificities of H-2 haplotypes of the inbred mouse strains. However, these reactions were only the consequence of the cross-reactions of anti-H-2 inbred sera with antigenic products of wild mice. In the set of wild mice tested, a frequent occurrence of the same H-2 phenotype in individuals originating from the same locality could not be confirmed.


Interaction of DNA and deoxyribonucleoprotein with methylene blue after the treatment with fixation solutions used in chromosome G-banding techniques. Maximum binding capacity of calf thymus DNA and deoxyribonucleoprotein for methylene blue was investigated. The binding capacity of native and heat-denatured solutions of the biopolymers was compared with that of the solutions incubated with fixatives used in cytology, viz. 4% formaldehyde and methanol-acetic acid (3 : 1) mixture. The binding capacity of DNA and DNP is strongly reduced after the treatment with both fixation agents. Thermal denaturation leads to a small decrease in the binding capacity of DNA, but does not influence that of DNP. These results were compared with microphotometric determination of nuclear methylene blue content in thymocyte spreads. Fixation with MeAc increased the dye content as compared with air-dried thymocytes, whereas 4% formaldehyde exhibited practically no influence. The mechanisms connected with the different staining properties after incubating with fixation agents DNA and DNP solutions on the one hand and the nuclear material of thymocyte spreads on the other hand are discussed.


Differentiation of rat lens epithelial cells in tissue culture. (I) Effects of cell density, medium and embryonic age of initial culture. The growth and differentiation of rat lens epithelial cells in tissue culture were studied. Cells could be maintained for a number of generations in an undifferentiated state in suspension culture. When cultured as monolayers, they grew and differentiated in a series of six defined stages described here. These stages include morphological changes (elongation, followed by cell "spreading" or formation of cell aggregates), and biochemical changes (appearance of nu-crystallin protein as detected by immunofluorescence). The process of differentiation appeared to be accelerated in the vicinity of elongated cells, occurred more rapidly at high cell density, and required frequent changes of medium. This suggests that cell-cell communication, and not medium factors, may be essential for promoting differentiation. The final morphology of the differentiated cells differed, depending on the embryonic age of the rats used as a source of lens epithelial cells. This implies that the programme for differentiation changes as a function of the embryonic age of the lens.


[Alpha 1 fetoprotein in pre-eclamptic toxemia (author's transl)]. From 1973 to 1975, 287 serum levels of alpha 1 fetoprotein in women with pre-eclamptic toxemia were determined. Pre-eclamptic toxemia was classified according to modified scheme of Goecke and Rippmann. 161 patients had mild pre-eclamptic toxemia (index 1-3), 72 patients had moderate pre-eclamptic toxemia (index 4-6), 54 patients had severe pre-eclamptic toxemia (index 7). In all types of severity of pre-eclamptic toxemia more levels of alpha fetoprotein were lower or higher than the normal levels including the standard deviations. The number of abnormal values rose with an increasing toxemia index. There was no statistically significant difference between too high values and too low values. Significantly more values were above and also below the normal values. Our investigations appear to indicate that the determination of the alpha fetoprotein is not only valuable as screening method for neural tube defects but also of value in the diagnosis and management of pre-eclamptic toxemia. Too high and too low values should not be differentiated but values both above and below the normal levels should be considered.


[Primary combined chemotherapy following removal of an endodermal sinus tumor or yolk-sac tumor of the ovary (author's transl)]. The yolk-sac tumor or endodermal sinus tumor is a rare but highly malignant ovarian teratoma with its highest incidence in young women. The tumor is relatively resistant to radiotherapy but sensitive to combined chemotherapy and should therefore be differentiated in the diagnosis from embryonal carcinoma and radio-sensitive dysgerminomas of the ovary. It is possible that the determination of alpha-fetoprotein is of differential diagnostic value. Two patients are alive and well 8 months and 2 1/2 years following removal of a yolk-sac tumor and primary chemotherapy. One patient received chemotherapy for peritoneal recurrence after removal of the tumor and radiotherapy. She died 1 1/2 years following the primary operation. Long term chemotherapy of the yolk-sac tumor is indicated following operation, irrespective of the stage of the tumor.


Activation of chymotrypsinogen by boar acrosin and its prevention by antiboar acrosin rabbit gamma-globulins. Activation of chymotrypsinogen by bovine trypsin or boar sperm acrosin was followed up using Nalpha-acetyl-L-tyrosine ethyl ester in a highly sensitive test system. Inhibition studies employing antiboar acrosin rabbit gamma-globulins showed the following results. 1) Whereas the acrosin-induced activation velocity was significantly depressed in the presence of the antibodies, the trypsin-catalyzed activation rate was not diminished. 2) The antibodies enhanced the acrosin-catalyzed cleavage rate of BzArgOEt significantly, but not the trypsin-catalyzed cleavage rate of this substrate. 3) Autodigestion of acrosin was considerably reduced in the presence of the antibodies. The enzymatic test system used is especially suitable to study the specificity of acrosin antibodies or their affinity to related enzymes if only small amounts of these substances are available.


Influence of boar acrosin antibodies produced in rabbit and sheep on chymotrypsinogen activation catalyzed by acrosin from boar, bull, ram, rabbkt and human. Activation of bovine chymotrypsinogen is catalyzed with increasing velocity by human, rabbit, boar, bull and ram acrosin. Antiboar-acrosin rabbit gamma-globulins cause a significant reduction in the proenzyme activation rate induced by boar and bull acrosin, but only a weak reduction or none if ram or rabbit acrosin is the activating agent. The antiboar-acrosin gamma-globulins from sheep strongly inhibit chymotrypsinogen activation by ram, bull and boar acrosin, and significantly inhibit the human acrosin-catalyzed reaction.


[On the alpha-fetoprotein in the diagnosis of primary liver carcinoma (author's transl)]. The existence of Alpha-Fetoprotein was investigated in sera of 544 patients with laparoscopically and/or histologically proved chronic liver diseases and cancer of the liver. The applied methods were the double-diffusion technique (micro-Ouchterlony), the counter immunelectrophoresis and the latex agglutination test. In 15 patients with primary cancer of the liver there was a positive result for Alpha-Fetoprotein with the first two methods, 73,3 and 80,67%, respectively. In other liver diseases (liver cirrhosis, chronic hepatitis, malignant tumors with liver metastases) there were only a few positive results (4,17% in patients with liver metastases of unknown origin and less than 1% for the other disease groups). In this group the latex test showed a higher frequency of positive tests for Alpha-Fetoprotein, however with a concomitant rise of positive Waaler-Rose test in 80-93%. In the case of a positive rheumatic factor the latex test will therefore demand a cautious judgement because the latex particle coated with rabbit-anti-Alpha-Fetoprotein-globulin could be agglutinated by anti-rabbit-globulin existing in human serum. A correlation between the existence of Alpha-Fetoprotein and clinical parameters or laboratory datas does not exist. Increases of Alpha-Fetoprotein shown by counter immunelectrophoresis exist in primary cancers of the liver and embryonic tumors, sometimes and only temporarely in acute viral hepatitis, in some sorts of liver cirrhosis of indian children and in the case of pathologic gestation. If these last-named diseases are excluded a positive test for Alpha-Fetoprotein by precipitation methods is highly suggestive for primary cancer of the liver. Finally, the determination of this substance has a certain value for the therapeutic control of Alpha-Fetoprotein positive malignant tumors.


Triggering of lymphocytes by antibodies against beta2 microglobulin. beta2 microglobulin (beta2m), structurally related to "domains" of immunoglobulin molecules, is associated with products of the major histocompatibility system on cell surfaces. Heteroantibodies against beta2m are mitogenic to a specific subpopulation of human and mouse B lymphocytes. This subpopulation is present in human blood which makes the antibody convenient and clinically useful as a functional marker for peripheral B lymphocytes. Absorption and elution experiments, as well as tests showing mitogenic activity of Fab monomers of anti-beta2m indicate that the interaction of the binding site of the antibody and relevant cell surface structures, probably beta2m itself, is directly responsible for lymphocyte activation. The relevance of these findings for cell receptors involved in lymphocyte activation is discussed.


Carcinoembryonic antigen-like substances in human cavity fluids. Pleural and peritoneal fluids from humans with pathological diseases were examined for the presence of carcinoembryonic antigen-like substances (CEA-LS). Among eight samples tested by a solid phase radioimmunoassay, two pleural fluids and one peritoneal fluid showed significantly elevated CEA-activity. The substances responsible for the CEA-activity were isolated by perchloric acid-extraction followed by two successive Sephadex G-200 chromatography into two pools, Pool I (PI) and Pool II (PII). According to their sedimentation properties, PII was slightly smaller than CEA from tumor tissue-extract (CEA-TTE), while PI was larger than CEA-TTE and approximately twice the size of PII. Micro-double diffusion and antibody binding studies showed that CEA-LS possessed identical antigenic determinants as CEA-TTE, which did not cross-react with normal colon antigen (NCA).


I. Effect of Trichinella spiralis infection on the migration of mesenteric lymphoblasts and mesenteric T lymphoblasts in syngeneic mice. The migration of [125I]UdR-labelled mesenteric lymph node cells in NIH strain mice at various times after inis produced an enhanced accumulation of mesenteric immunoblasts in the small intestine at 2 and 4 days after infection but not at later times. The enhanced migration occurred when using cells from both uninfected and infected donors, denoting an absence of antigenic specificity. This effect is not secondary to a reduced arrival of cells at sites away from the gut in infected mice, but to a primary increase of the arrival in the small intestine. Mesenteric T lymphoblasts (separated on a nylon-wool column) migrated to the small intestine of uninfected recipients and appear to be a major portion of the population which migrate to the gut of infected recipients. Our results were confirmed using 51Cr to label mesenteric cells. We conclude that the parasite causes the small intestine to become more attractive or retentive for mesenteric blast cells early during infection.


Study of human T and B lymphocytes with heterologous antisera. II. Partial characterization of a human T lymphocyte antigen. Using a horse anti-human thymocyte serum made specific for T lymphocytes by absorption (anti-HTLA serum), a material reacting with it has been extracted from detergent-lysed peripheral blood lymphocytes. Acid elution and gel-filtration analysis allowed recovery of two peaks containing HTLA activity: one of 30,000-40,000 Daltons, the other being excluded by Sephadex G-200 and likely to be more than 300,000 Daltons. HTLA appeared to be released by lymphocytes incubated at 37 degrees, but not at 4 degrees, and significantly detectable after 10-24-h incubation. Finally co-capping experiments with anti-beta2m antibodies and absence of inhibition of cytolysis by anti-HTLA serum of celld beta2m.


Fibroblast lysis by lymphocytes from normal persons and SLE patients on short-term cultures. Lymphocytes from thirty-six normal individuals of both sexes with ages ranging from 4-89 years were tested for their cytotoxic activity against short term cluters of fibroblasts from nine foetal umbilical cords and nine adult skin samples. Wide variability in the amount of fibroblast lysis was observed. Adult male lymphocytes caused significantly higher lytic activity than lymphocytes of adult females against both types of targets. Cell-mediated target cell reduction was not confined to allogeneic target cells. There was also killing of autochthonous cultured fibroblasts. Some individuals' sera inhibited the fibroblast lytic activity of their own lymphocytes. These cell-mediated reactions are directed against unknown antigenic specifications, probably not histocompatibility antigens, and give evidence of the problem of how to measure normal controls in the microcytotoxicity test in allogeneic human testing. Both autochthonous and allogeneic target cell lysis were also observed in SLE patients.


The physiochemical and immunological characterization of Gm (1) antibodies from normal human serum. The mol. wt and charge characteristics of Gm(1) antibodies from normal human serum were studied by gel filtration and DEAE- anion exchange chromatography. The effect on anti-Gm(1) activity, of incubating individual antisera with disulphide reducing agents, and with anti-IgG or anti-IgM immunoabsorbents were also studied. The results demonstrate the existence of a low molecular weight IgM protein with anti-Gm(1) activity.


Chromosomal considerations--application of modern cytogenetics. Attention is directed towards the use of new techniques in cytogenetics. C-, G-, Q-, and R-banding methods are briefly discussed and illustrated. Methods of sister chromatid differential staining and silver staining for demonstration of nucleolus organizing regions are reviewed and their application in medical cytogenetics is pointed out. Frequency of chromosomal aberration and the importance of the orofacial area in diagnosis of chromosomal syndromes are the bases for the argument for the usefulness of cytogenetics in the field of stomatology-dentistry.


Serum antibodies to herpesvirus early antigens in patients with cervical carcinoma determined by anticomplement immunofluorescence technique. Sera from patients with carcinoma of the uterine cervix were used with the anticomplement immunofluorescence (ACIF) technique to develop a simple procedure for detection of antibodies to herpes simplex virus type 2 (HSV-2) early antigens. Test cells used in the ACIF assay were HEp-2 cells infected with HSV-2 sequentially treated with inhibitors of protein and of DNA SYNTHESIS. The cells were first treated for 7 h with cycloheximide (100 mug/ml) and then for 3 h with hydroxyurea (150 mug/ml). In this assay serum titers giving more than 21.5% ACIF-positive cells correlated to invasive carcinoma of the cervix. Using these criteria 18 out of 43 patients with invasive carcinoma, none of 8 patients with carcinoma in situ, and only 1 of 43 controls, were diagnosed as positive. Thus an elevated titer of antibodies against HSV-2 antigens correlated to cervical carcinoma.


Antigens on melanoma cells detected by leukocyte dependent antibody assays of human melanoma antisera. Antibody-dependent cell-mediated cytotoxic assays have been used to examine antigens on human melanoma cells obtained either directly from patients or from long-term melanoma cell lines. A panel of melanoma antisera was selected from human subjects which could be shown not to have significant reactivity to histocompatibility antigens. With these antisera extensive cross-reactions between melanoma cells were found. However, the cross-reactivity was incomplete and the pattern of reactivity was different for each antiserum tested. These results were not consistent with a common melanoma antigen on human melanoma cells but rather indicated heterogeneity of melanoma antigens and multiple antibody specificities in the sera tested. This appeared to be confirmed by extensive cross-absorption studies which indicated limited cross-reactivity of antigens on melanoma cells from either long-term or short-term cultures. Several changes in the antigenic profile of melanoma cells in vitro from both long-term and short-term cultures were documented which resulted from contamination of the melanoma cell lines with non-melanoma cells and fibroblasts. Melanoma antisera may therefore be useful to mintor changes in long-term cultures which would otherwise give spurious results in in vitro tests. These results appear to have considerable significance for understanding tumour/host relationships and for the establishment of rational immunotherapeutic procedures and diagnostic tests in melanoma.


Serum alpha-fetoprotein as a marker for the effect of post-operative radiation therapy and/or chemotherapy in eight cases of ovarian endodermal sinus tumour. The clinical pathological findings of eight cases of ovarian endodermal sinus tumour (yolk sac tumour) are presented. Histological exmination in all eight cases showed a typical endodermal sinus tumour pattern, and in six of the patients other tumour elements such as dysgerminoma, choriocarcinoma, malignant teratoma, endometriosis, and a dermoid cyst were also found. Six patients had increased serum alpha-fetoprotein concentration in the post-operative period, and two patients had a normal concentration 27 and 35 days after operation, respectively. In all cases except one, a close correlation between serum alpha-fetoprotein and progression or regression of tumour was found. Serum alphafetoprotein was thus found to be a reliable parameter in post-operative radiation and/or chemotherapy (VAMBLE). In one patient who died 10 months after operation with widespread endodermal sinus tumour growth, only a small terminal increase in serum alpha-fetoprotein concentration was found. Four of the eight women are still alive with normal alpha-fetoprotein concentration, and without clinical evidence of tumour disease.