[Structure and histochemical characteristics of the mucosa of the Vater's ampulla in man (according to materials of aimed biopsies)]. Examination of 52 aimed duodenal biopsies has revealed that there are several variants of the structure of the Vater's papilla mucous membrane. In a number of cases it is identicalto the mucous membrane of the duodenum. In 20 cases the Vater's papilla was covered with mucoid epithelium which was histochemically similar to the epithelium lining the stomach, bile ducts and the gallbladder.  The "intermedial" forms found in these cases appearedto be capable to change into mucoid or edging epithelium. The mucous cells occasionally found in the Vater's papilla epithelium might be a variation of goblet cells. It seems that they are secreting by holocrine type.


[Receptors with circular nerve apparatus]. The observations conducted by Bielschowski-Gross method and intravital staining with methylene blue after A.S. Dogiel have shown that the receptors with an annular nervous apparatus are located in the muscular sheath, nerve bundles and ganglia of the intermusclar plexus of the oesophagus, stomach and small intertine of birds. For the first time it was shown taht the receptors with the annular nervous apparatus were not of the central origin  but might be formed by dendrites of the vegetative neurons of digestive tract.


[Morphological changes in the small intestine of rats after its temporary exclusion from blood circulation]. The experiments were performed in 56 albino rats (15of them served as a control. The small intestine was excluded from the blood circulation for 45minutes and then pieces of the proximal and  distal parts of the intestine were taken forhistological and histochemical study. It was shown that such a term of anemia resulted indeep but reversible changes in the mucous sheath and in the intramural nervous apparatus of thesmall intestine. The state of the organ wall became normal in 15 days after operation.


[Circadian rhythm of proliferative processes in the sebaceous glands]. The proliferation process indices (such as the mitotic activity 0/00,labelled cell 0/00 and the amount of mitoses arrested by colchicine 0/00) were studied in two sebaceous glands of rats-the gland of the external auditory meatus and the tarsal sebaceous gland. It was established that the diurnal changes of the number of mitotically dividing cells and the DNA synthesizing cells could be presented as one-peak curves, the maximum falling on the night and morning hours and the minimum-on the day-time and evening. The average diurnal mitotic activity in terminal portions was 16, 3-1, 9 and the average amount of labelled cells was 76, 9-6, 1. In the excretory ducts they were 13, 0-1,6 and 65, 6-5, 4 respectively. The amount of C-mitoses in rats which were given colchicine was 75, 7-5,1 in the morning hours and 58, 1-4, 5% in the evening hours. It permitted to calculate the duration of mitoses which were equal to 1, 9 hour in the morning and 1,2 hour in day-time.


Effect of experimental duck virus hepatitis infection on some biochemical constituents and enzymes in the serum of white Pekin ducklings. Experimental duck virus hepatitis infection of 11-day-old white Pekin ducklings having specific maternal antibodies revealed significant changes in some biochemical constituents and enzymes of the serum during the 3 weeks following exposure. These changes included a marked decrease in the total proteins and the albumin fraction, together with a significant elevation in levels of alkaline phosphatase, glutamic pyruvic transaminase, bilirubin, and creatinine. Most of these changes were attributed primarily to a deranged liver function associated with duck virus hepatitis infection.


[Bleomycin effect on DNA replication and proliferation kinetics using Ehrlich ascites tumor as a model]. The effect of the cytostatic agent bleomycin isolated from Streptomyces verticillus on DNA-synthesis and cell cycle of hyperidiploid Ehrlich-Lettré ascites tumor was studied in vivo and in vitro by means of liquid scintillation counting and pulse cytophotometry. A highly significant and dose dependent inhibition of 3H-thymidine incorporation into the tumor cells in vitro could be found. This inhibition of DNA-synthesis results in vivo in an initial decrease of cells in the S and (G9 plus M) phase in the DNA histogram. A partial synchronization of the tumor can be shown using 12.5, 25.0 and 50.0 mg/kg body weight of bleomycin. At a dose of 12.5 mg/kg body weight bleomycin, two subpopulations which pass through the cell cycle partially synchronized at a time interval of four hours, can be discriminated. This effect declines after the passage through one cell cycle. After 120 h most of the cells are in G1-area, another compartment in the late S-area and the smallest compartment in the (G2 plus M)-area of the DNA histogram. The arrest in the late S-area probably is the consequence of damage to the DNA synthetizing apparatus: DNA-synthesis cannot be completed and G2 cannot be reached. The significance of investigations concerning proliferation kinetics for tumor therapy is pointed out.


[Effect of ajmaline and its therapeutically used derivatives N-propylajmaline and di-monochloracetylajmaline on the functional refractory period and contractility of guinea pig atrium and aconitine arrhythmia in the rat]. 1. In the isolated left atrium of the guinea pig ajmaline and di-monochloracetylajmaline (DCAA) show almost the same activity concerning prolongation of the functional refractory period. 2. In contrast to this N-propylajmaline (NPA) is much more effective than ajmaline. 3. NPA as compared to ajmaline and DCAA, however, shows a considerably smaller difference between the concentrations prolonging refractory period (I) and those decreasing contractility (II) in the guinea pig atrium (EC25). The quotient from I and II is 0.4 with NPA, 1.2 with ajmaline and 1.6 with DCAA. 4. Differences in efficacy similar to those observed in the guinea pig atrium are also found in experimental cardiac arrhythmias in the intact animal. NPA is much more effective than ajmaline regarding inhibition of extrasystoles, ventricular tachycardia and ventricular flutter due to aconitine infusion in the rat. In this experimental model DCAA shows slightly less activity than ajmaline; this difference is statistically significant.


"A" protein of amyloidosis. Isolation of a cross-reacting component from serum by affinity chromatography. A nonimmunoglobulin protein (A protein) has been isolated from amyloidotic tissue of secondary type. Antisera prepared to this protein identified a cross-reacting substance in the sera of patients with secondary amyloidosis. Sera from 70 persons with amyloidosis, 120 normal adults, 20 aged persons, and 97 patients with chronic diseases were tested for this substance. One hundred percent of secondary amyloid sera had amounts of amyloid serum component detectable by double diffusion in agar, whereas only 19 percent of primary amyloid sera were positive. Approximately 60 percent of rheumatoid sera as well as 60 per cent of sera from aged individuals were positive. Only 3 percent or normal blood donors had detectable amounts of this circulating substance. Isolation of the serum component by affinity chromatography and partial characterization have shown that it is an alpha-globulins with a molecular weight of 100,000-120,000, that it is not related antigenically to immunoglobulin or amyloid P-compoment, and that it has an amino acid analysis that is markedly different from tissue A protein. The possible participation of this substance in the genesis of amyloid is discussed.


Hydralazine therapy in hypertensive patients with idiopathic systemic lupus erythematosus. Seven hypertensive patients with idiopathic systemic lupus erythematosus were treated with hydralazine. They received a mean daily dose of 203 mg for a mean duration of 21 months. All were taking prednisone alone or in combination with azathioprine. During therapy with hydralazine, there were no new symptoms nor exacerbation of pre-existing symptoms attributable to systemic lupus. Laboratory parameters, including antinuclear antibody titers and complement levels, either improved or remained stable. The results indicate that hydralazine can be safely used in hypertensive patients with systemic lupus who are receiving concomitant immunosuppressive therapy.


Changes in sensitivity to radiation and to blemycin occurring during the life history of monolayer cultures of a mouse tumour cell line. The response to X-radiation and to bleomycin has been measured at a number of times during the life of monolayer cultures of EMT6 mouse tumour cells. Little change in radiation sensitivity was seen at any time and no loss of the shoulder to the survival curve occurred. Cultures in early plateau phase (where a considerable amount of cell proliferation is balanced by cell loss) showed a reduced sensitivity to bleomycin when compared with cells in exponential growth. However, after a longer period in plateau phase, when proliferation had virtually ceased, the sensitivity became greater than that of exponetial phase cells. These findings are discussed with reference to the conflicting results of other workers.


Carcinoembryonic antigen: characterization of binding with insoluble lectins. Insoluble conanavalin A and wheat germ agglutinin (WGA) were found to bind to carbohydrates on radio-labelled carcinoembryonic antigen (CEA). Binding by WGA was inhibited both by N-acetyl D-glucosamine and fragments of antibody to CEA, but was increased by intact antibody to CEA. This suggests that WGA binds to exposed N-acetyl D-glucosamine determinants on 125I CEA and also on antibody molecules. It also suggests that 125I CEA contains binding sites for anti-CEA which contain N-acetyl D-glucosamine as well as others which do not. Molecules of 125I CEA which bound to the insoluble lectins were more antigenic for anti-CEA than unbound molecules. These results suggest that the principal antigenic site on CEA contains N-acetyl D-glucosamine and may help to explain the agglutination of tumour cells by lectins.


Immunity to murine sarcoma virus induced tumours. IV. Direct cellular cytolysis of 51Cr labelled target cells in vitro and analysis of blocking factors which modulate cytotoxicity. The antigen specific cell mediated cytotoxicity of MSV immune spleen lymphocytes to 51Cr labelled murine lymphoma cells was wholly abolished by pretreatment of the spleen cells with anti-theta antibody and complement. Early during the immune response to MSV the cytotoxic acitivity was inhibited by incubation of immune lymphocytes with "late progressor" or "early regressor" serum. Immune lymphocytes at later times were more refractory to such inhibition by serum blocking factors. Although unfractionated cytotoxic lymphocytes, irrespective of the time after MSV infection at which they were tested, were inhibited by soluble tumour associated antigen (TAA), a subpopulation of cytotoxic T cells was identified which was inhibited neither by antigen nor serum.


Studies on mouse Moloney virus induced tumours: I. The detection of p30 as a cytotoxic target on murine Moloney leukaemic spleen cells, and on an in vitro Moloney sarcoma line by antibody mediated cytotoxicity. Antigenic determinants of p30, the most abundant internal virion protein of C type RNA viruses, were detected on the surface of spleen cells from mice bearing Moloney leukaemia and on an in vitro line of Moloney sarcoma, MSC. On both cell types, these determinants on the p30 molecules served as cytotoxic targets in a xenogenic complement dependent antibody mediated 51Cr release assay. Two antisera were used: a rat anti MLV -M induced lymphoma serum, and an antiserum raised in goats to either disrupted FeLV. The cytotoxic target antigens of these antisera were analysed by inhibition of cytotoxicity with viral and cellular proteins.


Studies on mouse Moloney virus induced tumours: II. Detection of p30 in the serum of mice with Moloney leukaemia by in vitro blocking of complement dependent antibody mediated cytotoxicity. Sera from Balb/c mice bearing Moloney leukaemia block complement dependent antibody mediated cytotoxicity of an antiserum prepared in rats against syngeneic Moloney virus induced lymphomata when either spleen cells from mice bearing Moloney leukaemia (M) or an in vitro line of Moloney virus transformed cells (MSC) are used as targets. This antiserum has been shown to recognize p30, the major internal virion protein, as a cytotoxic target on these cells. Viral particles were identified by electron microscopic examination of pelleted material obtained from leukaemic sera after high speed centrifugation. However, removal of virus did not affect the capacity of the leukaemic sera to absorb cytotoxicity of rat ILR-3 for MSC targets, and only depressed somewhat its ability to absorb activity of the same antisera against M targets. Virus-free leukaemic sera also blocks complement dependent antibody mediated cytotoxicity of an antiserum prepared in goats against the gs3 determinant of p30. This indicates that the material in leukaemic sera responsible for the in vitro block of antibody mediated cytotoxicity was p30. A lesser degree of block was observed with sera obtained from normal Balb/c mice, but the nature of material responsible is as yet undefined.


Study of the antigenic cross reactivity between carcinoembryonic antigen and "nonspecific cross reacting antigens" (NCA and NCA 2). The immunochemical relationship between CEA, NCA and NCA 2 was studied in guinea-pigs. Strong cross reactions were found between these antigens, either in delayed or anaphylactic reactions. Some specific determinants for each antigen could still be demonstrated. Delayed hypersensitivity is likely to be due to the protein moiety of the molecules while anaphylactic reactivity could probably be related to their glucidic parts. Consequently, CEA and NCA have common antigenic determinants on their glucidic and peptidic moieties, perhaps more on the latter ones.


Cellular immunity to myelin basic protein in man and in animal model systems as measured by the macrophage migration inhibition test. Lymphocytes from patients with neoplastic disease were tested for sensitization to encephalitogenic factor (EF) by the macrophage migration inhibition test. Sensitization to EF was demonstrated in 71% of patients with various forms of neoplastic disease. Sensitization to EF was also demonstrated for 31% of subjects with no evidence of neoplastic disease; these included patients with warts, chronic bronchitis and hernias. In contrast, healthy subjects showed no sensitization to myelin basic protein. These observations suggest that sensitization to EF may not be confined to patients with neoplastic disease. Lymphocytes from hamsters bearing a transplanted virus induced tumour were sensitized to EF prepared from both human and hamster brain. Sensitization was also seen in hamsters infected with influenza virus but not in animals with acute tubular necrosis produced by glycerol treatment. The development of an animal model system provides a method for the investigation of possible mechanisms of sensitization.


Differences in proliferative activity of rat and human prostate in culture. The properties of human benign prostatic hyperplasia (BPH) and rat prostate were compared after culture in the absence of insulin and testosterone. Quantitative methods were used to assess changes in tissue composition and the height of the epithelial cells. BPH appeared less sensitive than rat prostate to withdrawal of hormone support, and the changes which occurred during culture of BPH were more typical of a repair mechanism to injury than of a castration effect. Cell kinetics was investigated using [125I] iododeoxyuridine and vincristine. Both approaches demonstrated a spontaneous surge in proliferative activity of BPH reaching a peak at about Day 4. In contrast, proliferative activity in rat prostate tended to fall over the period of 2-8 days of culture. The significance of these findings in terms of age linked effects is discussed.


The interaction of Concanavalin A and the surface coat of stratified squamous epithelium. The interaction of Concanavalin A (Con A) and the surface membranes of the epithelial cells of human oral mucosa, epidermis and guinea-pig lip was studied. Two methods were used: fluorescent Con A and Con A in combination with horse radish peroxidase. With both methods the surface coat of epithelial cells was stained. The specificity of the staining methods could be demonstrated. The interaction between pemphigus antibodies and the surface membranes of epithelial cells could be blocked by Con A.


Light-induced leucine transport in Halobacterium halobium envelope vesicles: a chemiosmotic system. Halabacterium halobium cell envelope vesicles accumulate L-[14-C]leucine during illumination, against a large concentration gradient. Leucine uptake requires Na-+ and is optimal in KCl-loaded vesicles resuspended in KCl-NaCl solution (1.5 M:1.5 M). Half-maximal transport is seen at 1 X 10-minus 6 M leucine. In the dark the accumulated leucine is rapidly and exponentially lost from the vesicles. The action spectrum and the light-intensity dependence indicate that the transport is related to the extrusion of protons, mediated by bacteriorhodopsin. Since light gives rise to both a pH gradient and an opposing transmembrane potential (interior negative), it wass responsible for providing the energy for leucine transport. The following results were obtained under illumination: (1) membrane-permeant cations and valinomycin or gramicidin greatly inhibited leucine transport without altering the pH gradient; (2) buffering both inside and outside the vesicles eliminated the pH gradient while enhancing leucine transport; (3) dicyclohexylcarbodiimide increased the pH gradient without affecting leucine transport; (4) arsenate did not inhibit leucine uptake. A diffusion potential, established by adding valinomycin to KCl-loaded vesicles, caused leucine influx in the dark. These results suggest that the leucine transport system is not coupled to ATP hydrolysis, and responds to the membrane potential rather than to the pH gradient. The Na-+ dependence of the transport and the observation that a small NaCl pulse causes transient leucine influx in the dark in KCl-loaded vesicles, resuspended in KCl, even in the presence of p-trifluoromethoxycarbonylcyanide phenylhydrazone or with buffering, suggest that the translocation of leucine is facilitated by symport with Na-+.


Tyrocidine and the linear gramicidin. Do these peptide antibiotics play an antagonistic regulative role in sporulation? 1. The cyclic peptide antibiotic tyrocidine, synthesized by Bacillus brevis (ATCC 8185), inhibits RNA synthesis in an in vitro transcriptional system by forming a complex with the DNA. 2. The linear peptide antibiotic gramicidin, synthesized by the same strain, reverses at least partly this inhibition. The molecular mechanism of this reactivation is unknown. Gramicidin by itself inhibits transcription in vitro. This inhibition is not due to a complex formation between DNA and the peptide. 4. A possible regulative role of the two peptides in sporulation is discussed.


Reduced nicotinamide adenine dinucleotide dependent reduction of fumarate coupled to membrane energization in a cytochrome deficient mutant of Escherichia coli K12. Escherichia coli SASX76 does not form cytochromes unless supplemented with 5-aminolevulinic acid. It can grow anaerobically on glycerol and DL-glycerol 3-phosphate in the absence of 5-aminolevulinic acid with fumarate but not with nitrate as the terminal electron acceptor. Cytochrome-independent NADH oxidase, glycerol 3-phosphate- and NADH-fumarate oxidoreductase activities are induced by anaerobic growth on a glycerol-fumarate medium. The pathway of electrons from substrate to fumarate involves menaquinone. The NADH-fumarate oxidoreductase and cytochrome-independent NADH oxidase systems are inhibited by piericidin A, 2-heptyl-4-hydroxyquinoline N-oxide, and iron chelating agents. Both systems can energize the membrane particles as indicated by quenching of atebrin fluorescence.


The characterization of multiple forms of kynurenine formidase in Drosophila melanogaster. Two enzymic forms of kynurenine formamidase (EC 3.5.1.9) from Drosophila melanogaster were separated and partially purified by pH fractionation, (NH4) 2SO4 fractionation and Sephadex G-75 gel filtration. The enzymes were also separated by DEAE-cellulose ion-exchange chromatography and distinguished by their different rates of thermal inactivation. The multiple forms are termed formamidase I and formamidase II. The molecular weight of formamidase I as measured by Sephadex G-75 chromatography is 60 000 and that of formamidase II is 31 000. The pH optima are broad, ranging between 6.7 and 7.8 for formamidase I and 6.5 and 8.0 for formamidase II. The apparent Km values are 5-10(-3) and 0.83-10(-3) M, resepctively. The possibility that formamidase II is an active subunit of formamidase I is discussed, although neither enzyme will convert to the other when separated and rechromatographed. Eight organisms were tested for the presence or absence of multiple forms of formamidase. Drosophila melanogaster and Drosophila virilis have both enzymes; cow, chicken, yeast and housefly have formamidase I only, and mouse and frog have formamidase II only.


Localizing -SH groups in monoaminergic synaptic vesicles with the mixture of zinc iodide-osmium tetroxide (ZIO). In monoaminergic granulated vesicles the mixture of zinc iodide-osmium tetroxide (ZIO) stains two compartments: the core and the matrix, the latter being more intensively stained than the core. Rat pineal glands were incubated in 0.1 M N-ethylmaleimide (NEM). It was observed that NEM blocks ZIO reaction both in the matrix and the core, whereas the controls were fully reactive. This finding strongly suggests that ZIO reaction is due to -SH groups and can be correlated with the effect of NEM on uptake processes.


Studies on the cerebellar projections from the main and external cuneate nuclei in the cat by means of retrograde axonal transport of horseradish peroxidase. The cerebellar projections from the main and external cuneate nuclei in the cat have been studied by means of retrograde axonal transport of horseradish peroxidase. The main projection from the external cuneate nucleus (ECN) is to the intermediate and, possibly, the small lateral part of lobule V and to the paramedian lobule on the ipsilateral side. The projection from the ECN to the cerebellar regions mentioned is topographically organized. Cells in the caudal part of the ECN send their axons to the caudal parts of lobule V and to the rostral part of the paramedian lobule. Cells in the rostral part of the ECN project to the rostralmost part of lobule V and to the folia in the caudal part of the paramedian lobule. The experimental study also shows that cells in the main cuneate nucleus (MCN) send their axons to the cerebellum. These axons, like those from the ECN, terminate in the intermediate part of lobule V of the anterior lobe and in the paramedian lobule. However, the axons of the cells in the MCN terminate only in the superficial parts of the folia, whereas those from the ECN terminate in the depth of the folia in these two cerebellar areas. The present study also gives evidence that cells in the ventral part of the gracile nucleus send their axons to lobules I and II of the anterior lobe vermis. The observations referred to here are to our knowledge the first anatomical findings demonstrating a projection from the main cuneate and gracile nuclei onto the cerebellar cortex. The observations confirm previous physiological studies.


Bleomycin combination chemotherapy in the management of testicular neoplasia. Eighty-three patients with Stage II or Stage III germinal neoplasia of the testis and 7 patients with extragonadal primary tumors were treated with bleomycin plus vinblastine, or a five-drug program, bleomycin plus cyclophosphamide, vincristine, methotrexate, and 5-fluorouracil. Of the 70 Stage III patients, there were 53 responses (75%), 22 complete and 31 partial. The mean survival of the complete responders is 100+ weeks, with 3 dead. The mean survival of the partial responders and nonresponders is 38 weeks and 33 weeks, respectively. There is a highly significant difference between complete responders vs. partial and nonresponders (p less than 0.01). Thirteen patients with nonmeasurable disease (Stage II and Stage III postresectional status) but at great risk to develop widespread metastasis were treated prophylactically after conventional therapy. Nine continue in complete response to 36 months. The 7 extragonadal primary patients showed 4 partial responses, none complete. Major toxicity was myelosuppression and also bleomycin pneumonitis in 5 of the 90 evaluable patients.


COMB (cyclophosphamide, oncovin, methyl-CCNU, and bleomycin): a four-drug combination in solid tumors. One hundred eighty-nine patients received a four-drug combination consisting of cyclophosphamide, Oncovin (vincristine), methyl CCNU, and bleomycin (COMB), according to three different drug regimens, performed sequentially. Of the 189, 62 had a partial response (33%) including 11/33 with squamous lung cancer, 11/32 with squamous carcinoma of the head and neck, 13/15 with oat cell carcinoma of the lung, and 7/41 with malignant melanoma. The response rate for patients with squamous lung or head and neck cancer appeared to be higher at weekly bleomycin doses of 30 and 60 mg (15/33 = 45%), compared to a weekly bleomycin dose of 15 mg (7/32 = 25%). A median survival from treatment of 30 weeks was observed in oat cell carcinoma, which represents considerable prolongation over that expected from supportive care alone or single-agent chemotherapy. Toxicity included: 1) myelosuppression, resulting in hospitalization for antibiotics in 20% of patients; 2) probable bleomycin lung damage in 4% of patients; and 3) dose-limiting vincristine neuropathy in 11%. The combination of twice-weekly vincristine and bleomycin for more than 6 weeks produced a disturbing "debilitation syndrome," characterized by weakness, anorexia, weight loss, and apathy. The encouraging response rate suggests a future role for these drugs in combination, especially for vincristine and bleomycin, with other agents showing activity in squamous and oat cell carcinoma. Toxicity precludes recommendation of this combination, in the regimens tested, for broader Phase III studies.


Nodular lymphoma: bone marrow and blood manifestations. Morphological and clinical features of 39 patients with nodular lymphoma were studied with particular reference to bone marrow and blood involvement. Twenty-one patients (54%) were found to have bone marrow involvement at the time of initial diagnosis. Thirteen (33%) also had peripheral blood involvement. A specific cell type, the small nodular lymphoma cell, was found in the bone marrow smears of 19 and the 21 patients with bone marrow involvement and in the peripheral blood smears of all patients with blood involvement. When large cells predominated in the lymph node or bone marrow sections, the bone marrow smears and the imprints of the lymph node and trephine biopsy demonstrated these large cells to have morphological qualities of lymphocytes rather than histiocytes.


Comparison of alpha1 fetoprotein radioimmunoassay method and liver scanning for detecting primary hepatic cell carcinoma. Alpha1 (alpha 1) fetoprotein (AFP) radioimmunoassay method was routinely used in addition to liver scintigraphy to detect a primary hepatic cell carcinoma, and the diagnostic accuracy of both methods was compared. Twenty-one of 27 cases (78%) with primary hepatic cell carcinoma showed a positive AFP titer of over 200 ng/ml. In 3 of these AFP-positive cases, no focal defects could be found in liver scintigraphy, although subsequently performed celiac angiography revealed hypervascular shadows. On the contrary, 22 of 27 cases (81%) represented well-defined focal defects on scintigraphy. In 4 of these cases with a positive scan, the result of AFP was found to be negative. The overall diagnostic accuracy for detecting primary hepatic cell carcinoma with a combination of both methods was 93%.


Bony metastases from carcinoma of cervix. Occurrence, diagnosis, and treatment. Fifty-five patients treated for cervical carcinoma developed bony metastases between January 1, 1961 and December 31, 1973. Roentgenograms were diagnostic in all but 2 of the patients. In 15 patients, a combination of radioactive scans and roentgenograms was used to establish the diagnosis. The most common mechanism of bony involvement from carcinoma of the cervix was extension of the neoplasm from para-aortic nodes, with involvement of the adjacent vertebral bodies. The earliest metastases were discovered at the time of the primary diagnosis. Thirteen years was the longest interval from the primary diagnosis until the discovery of bony metastases. Sixty-nine percent of the patients were diagnosed within 30 months. Ninety-six percent of the patients died within 18 months. Seventy-six percent of the patients received some form of therapy for their metastases. Thirty-six of these patients were treated with radiation therapy. Four of these patients received complete relief of symptoms, 24 some relief, and 8 patients received no relief.


An unusual leiomyosarcoma of the uterus containing osteoclast-like giant cells. An unusual variant of a poorly differentiated leiomyosarcoma of the uterus, containing osteoclast-like giant cells, is described. Areas of the resected tumor bore a close resemblance to giant cell tumor of bone. It is postulated that these giant cells, as well as the osteoclast-like giant cells reported in a number of tumors of other tissues, originate from the monocyte/histiocyte group of cells. The diagnosis of leiomyosarcoma was possible only after the tumor had been examined in the electron microscope. The ultrastructural features are consistent with a smooth muscle origin of the neoplasm; the final assessment is based on the relative proportions and frequency of such structures as: bundles of myofilaments with focal densities, marginal densities, pinocytotic vesicles, and an external lamina.


Inverted papillomas of the urinary bladder. Study of 20 patients with inverted papilloma of the urinary bladder and urethra revealed that the lesion is a true neoplasm, benign in its histologic morphology and clinical behavior. The lesion is believed to arise from the trigone and bladder outlet as a result of chronic proliferative cystitis. It occurs predominantly in men who are past middle age. The most commonly associated clinical symptoms are hematuria and those of urinary obstruction. The lesion may be easily mistaken for a low-grade papillary transitional cell carcinoma, although the histologic appearance is distinctly different, as is its subsequent behavior.


Acinic cell carcinoma of the parotid salivary gland presenting as a cyst. Report of two cases. Two case reports of pure cystic acinic cell carcinoma of the parotid gland are presented, together with a review of the literature. Both cases showed some departures from the classical histologic features of acinic cell carcinomas, particularly in reference to the intracystic papillary components. Pure cystic acinic cell carcinomas of salivary gland are a rarity in the literature and may appear histologically and morphologically innocuous on superficial examination.


The role of chemotherapy in the management of cancer of the head and neck: a review. Although experience with drug therapy of advanced or recurrent squamous cell carcinoma of the head and neck is limited, several agents have produced convincing and reproducible tumor regression in these patients. Methotrexate has had the widest usage, and produces 30-50% response rates; bleomycin, hydroxyurea, and adriamycin appear to be somewhat less effective. Location of the malignancy and previous x-ray treatment appear to be important determinants of responsiveness to methotrexate, while degree of differentiation has not yet been shown to be an important factor for response to this drug. Attempts to improve the response rate and duration of chemotherapeutic response by utilizing combinations of drugs, or use of drugs to sensitize the tumor to x-ray treatment, or to reduce the bulk of tumor before x-ray treatment, are reviewed; they have been only moderately encouraging. Intra-arterial chemotherapy appears to have a therapeutic advantage over intravenous treatment; however, the morbidity associated with the former approach limits its usefulness for routine usage. The use of drugs as adjuncts following surgery and/or radiation therapy or immunotherapy are newer approaches that have not been investigated sufficiently, but are promising areas for investigation.


Effects of nitrosocarbaryl on BALB/3T3 cells. Carbaryl(N-methyl-1-naphthylcarbamate) and its nitrosated product, N-nitrosocarbaryl, were tested for their effects of BALB/3T3 (clone A31) cells in culture. Nitrosocarbaryl, but not carbaryl, caused transformation of the BALB/3T3 fibroblasts, but neither chemical induced the complete expression of endogenous murine leukemia virus. Transformed cells differed from the parental control cells by loss of contact inhibition, change in morphology, growth in soft agar, growth to higher saturation densities, and tumorigenicity in normal newborn and irradiated weanling mice and athymic (nude) mice. Transformed clones were found to be negative for expression of RNA tumor virus antigens, viral reverse transcriptase, and infectious virus. Thus, it appears that nitrosocarbaryl can transform BALB/3T3 cells to tumorigenic cells with altered biological properties but without complete activation of RNA tumor viruses in the transformed cells. Expression of viral antigen in the transformed cells was inducible by iododeoxyuridine, indicating that the endogenous viral genome was retained in an unexpressed state.


Tumor immunoprophylaxis in mice using glutaraldehyde-treated syngeneic tumor cells. The ability of glutaraldehyde-treated tumor cells to induce protection against subsequent challenge has been studied in a syngeneic system. Two tumors have been tested in BALB/c mice. The first was a methylcholanthrene-induced tumor that has been maintained in serial passage for over a decade. The second was a spontaneous mammary adenoacanthoma that was tested at the third passage. The protection was found to consist of two components: (a) a specific immunological component; and (b) a nonspecific component observed when the immunizing and challenge dose were both given i.p. This nonspecificity, while possibly an element in tumor protective mechanisms, may confuse careful analytical studies of the immunogenic potential of tumor antigens.


The effects of bleomycin on immunocompetence in man. Bleomycin was administered to six patients with advanced cancer. Multiple parameters of both antibody- and cell-mediated immunity were followed serially to characterize the effects of bleomycin on immunocompetence in humans. Antibody-mediated immunity, including primary vaccination with keyhole limpet hemocyanin, was not depressed. While there was no significant suppression of cell-mediated immunity, phytohemagglutinin-stimulated lymphocyte blastogenesis was reduced after treatment with bleomycin. The in vitro effects of bleomycin on lymphocyte stimulation were studied, and while thymidine incorporation was significantly inhibited by bleomycin, leucine incorporation was not reduced even at high concentrations of bleomycin. We have concluded that bleomycin does not suppress immunocompetence in man.


Atopic hypersensitivity to cis-dichlorodiammineplatinum(II) and other platinum complexes. Allergic reaction to an antitumor agent, cis-dichlorodiammineplatinum(II) (DDP) was investigated. A 15-year-old white male with pulmonary metastases from embryonal carcinoma of testis was treated with a combination of DDP, bleomycin, and vinblastine. The dose of DDP varied from 2 to 2.25 mg/kg given i.v. He received 7 doses of DDP in 9 months. An anaphylactic reaction was seen within 3 min of the initiation of i.v. infusion of the 8th dose of DDP. The reaction was due to atopic hypersensitivity, as confirmed by an immediate wheal and flair reaction and increased histamine release from leukocytes with DDP. His serum IgE level was elevated. Neither the presence of chloride nor the amine grouping in DDP was essential for reactivity. The replacement of platinum with palladium abrogated the reactivity. There was no cross-reactivity with 3 other platinum complexes of known antitumor activity (platinum blue, platinum(II) 1,2-diaminocyclohexane malonate, and platinum(II) ethylenediamine malonate). This was also confirmed by the lack of reaction to subsequent i.v. administration of platinum(II) 1,2-diaminocyclohexane malonate (10 mg/kg) in this patient.


In vitro biochemical and cytotoxicity studies with 1-beta-D-arabinofuranosylcytosine and 5-azacytidine in combination. The effect of 1-beta-D-arabinofuranosylcytosine (ara-C) and 5-azacytidine (5-aza-C), alone and in combination, on DNA synthesis and cytotoxicity in hamster fibrosarcoma cells has been studied. After a 2-hr exposure of S-phase cells to ara-C at concentrations of 2 to 200 muM, the cells required about 4 to 6 hr to recover from inhibition of DNA synthesis. When 2 exposures to ara-C were used, maximal cytotoxicity occurred when the 2nd dose of ara-C was administered at the time when the cells recovered from the inhibition of DNA synthesis. When the S-phase cells were exposed to ara-C, the maximal killing effect of 5-aza-C occurred when this agent was administered 6 hr later, at the time when the cells had recovered from the inhibition of DNA synthesis. When S-phase cells were exposed to 5-aza-C, the maximal cell kill produced by ara-C also occurred 5 to 6 hr later. When the S-phase cells were exposed simultaneously to both ara-C and 5-aza-C, significant antagonism with respect to cytotoxicity was observed between these 2 agents. When cells in G1 were exposed to 5-aza-C, the cytotoxicity produced by ara-C on these cells when they entered S phase was additive with respect to the cytotoxicity produced by 5-aza-C exposure alone.


Partial characterization of C-type particles in a cell line (WR-9) derived from a rat epidermoid carcinoma of spontaneous origin. A C-type virus continuously released from a cell line (WR-9) derived from a spontaneous epidermoid carcinoma was purified by means of large-scale tissue culture techniques and high-volume zonal centrifuges. With the use of relatively pure virus concentrates, partial characterization of the virus has been accomplished. Up to 60 liters of spent culture medium from relatively low virus-yielding cultures were processed at a time through the Model K ultracentrifuge in order to obtain quantities of virus sufficient for convenient Tween-ether extraction of the major polypeptide (30,000 daltons). This structural protein having group-specific reactivity was purified and isolated by isoelectric-focusing techniques. A UV absorption peak (A280) was found to be coincident with a major peak of radioacticity at pH 8.6, the isoelectric point (pI) for rat virus gs antigen previously reported by other investigators. Because species-specific (gs-1) and cross-reactive (gs-3) determinants coexist on this protein, fractions containing the group-specific antigen were identified on the basis of the mammalian interspecies determinant (gs-3), using antiserum prepared against Tween-ether-disrupted feline leukemia virus. At the same time, reactivity to the gs-1 determinants in identical fractions was observed in complement fixation and gel diffusion assays, using guinea pig antiserum known to contain principally antibodies to rat gs-1 determinants. Presently, the principal source of rat type C viral gs antigen is rat cell line MSB, which continuously releases a rat leukemia virus pseudotype of murine sarcoma virus. The WR-9 rat virus line may be of use in providing an additional source of C-type particles that are capable of yielding good gs reagents.


[Presence of the H blood group antigen on a carcinoembryonic antigen (CEA). Enzymatic transformation of H to A and B specificities]. The carcinoembryonic antigen (CEA-M) was purified from a hepatic metastasis obtained from a blood group O patient with a cancer of the rectum. Using 125I-labelled-CEA and blood group antisera, H specificity was found on the CEA-M; the addition of anti-H to anti-CEA does not modify the binding of labelled-CEA-M to its antibodies (86%), this result leads us to conclude that H and CEA determinants are carried by the same molecule. However the low percentage of binding (30% with 1/10 anti-H) suggests that only a few CEA-M molecules do carry the H antigenic determinant. Finally, glycosyltransferases were used to modify the H specificity into blood group A and B specificities.


[The junctions of the sinoatrial fascicle in frogs]. The demonstration of gap-junctions on frog atria muscle similar to those described on mammalian myocardium has been controversed. En bloc staining with uranyl acetate allows us to show such gap junctions on this tissue.


[Changes in the level of alpha1-macroglobulin in rats during Masugi nephritis]. Using chromatographic technique on "Sephadex G 200" associated to immunologic qualitative and quantitative techniques, the authors showed that the alpha1-macroglobulin is present in normal Rat urines. This protein is also present in urines obtained during the Masugi nephritis. The alpha1-macroglobulin concentration observed in the different urine samples is unrelated to the total proteinuria evolution. The results given allow us to discuss the mechanisms of the glomerular filtration.


[Immunosuppressive effect of a human serum ferroprotein of hepatic origin, alpha 2 H globulin. Study on blastic transformation of normal lymphocytes in the presence of phytohemagglutinin]. The alpha2 H globulin, a glycoferroprotein, has been found in the serum of patients with malignant diseases. Its level varies according to the evolution of the disease. The activity of this protein has been studied on the normal lymphocytes cultivated in presence of phytohemagglutinin. This study has showed that 5 mug/ml of alpha2 H globulin can inhibit the blastic transformation. The inhibitory effect is proportional to the amount of alpha2 H globulin added as judged by the tritiated thymidin incorporation. The study of morphological aspects of the lymphocytes shows that alpha2 H globulin acts upon the synthesis of the cytoplasmic proteins.


The postponed compensatory pause as a manifestation of positive feedback in atrioventricular conduction. A patient with hypertensive cardiovascular disease was found to have unusual varieties of premature atrial and ventricular contractions. If the premature atrial contraction resulted in a greatly prolonged P-R interval, such that the increment in P-R exceeded the decrement in the preceding R-P, the next sinus P wave, occurring after a normal P-P interval, was found to be blocked. Also, numerous interpolated ventricular extrasystoles were observed in which the postextrasystolic P-R intervals were markedly prolonged and in which the compensatory pauses were postponed for one or two beats. We also present data from one dog in which a premature atrial activation produced a chain reaction such that complete A-V block occurred three beats later. We propose that the chain reaction which evoked the delayed block in the dog and the postponed compensatory pauses in the patient reflects the operation of a positive feedback mechanism in A-V conduction. Positive feedback is initiated by an extremely long P-R, which results in a very short R-P before the next cycle. This then leads to a still longer P-R, which then elicits a still shorter R-P. Block ultimately supervenes when the atrial activation wave arrives at the A-V junction during its effective refractory period.


De subitaneis mortibus. XIV. Bacterial arteritis in Whipple's disease. Although Whipple's disease is clinically recognized for its features of arthritis and diarrhea, pericarditis and cardiac valvular disease are frequently present and a significant number of such patients die suddenly and unexpectedly. This report includes special postmortem cardiovascular examinations in a 55-year-old man who died of Whipple's disease. Pericarditis and valvular disease were extensively present. There was also focal myocardial degeneration, including the sinus node, A-V node and His bundle. Typical Schiff-positive bacilli were found in the tunica media and endothelium most numerously in the small coronary and hepatic arteries, less in splenic and renal arteries, and least in small pulmonary arteries. Large coronary arteries and the aorta were not involved. Three stages of bacterial invasion of the arteries included a noninflammatory phase in which the bacilli were most numerous, a pancreatic phase in which bacilli were distinctly less numerous, and a healed scarring of arteries previously damaged. The combination of pericarditis, valvular disease of the heart, coronary and systemic bacterial arteritis and focal myocardial degeneration and myocarditis is unusual for Whippl'e disease. Studies of other cases are warranted to determine whether bacterial arteriopathy and arteritis have previously been overlooked or if the present case is unique. Evidence that the conduction system of the heart may be involved indicated that careful attention to cardiac rhyth and conduction is merited in future studies of patients with Whipple's disease.


The etiology of osteosarcoma. A review of current considerations. Various agents have caused osteosarcoma in several experimental animal systems. These agents or initiators may be classified as chemicals, radiation, viruses, and miscellaneous. Zinc beryllium silicate with beryllium oxide in rabbits and FBJ virus in mice are two such initiating agents. The relevance of these animal experiments to the human situation is not known, but recent reports regarding a transmissible agent obtained from human osteosarcoma tissue suggest that a virus may be implicated. There is a theoretic indication that the various etiologic agents, including viruses, may affect the DNA of normal cells in such a way that further evolution and differentiation through several cell divisions may result in the clinical appearance of cancer.


Endogenous primate and feline type C viruses. 1. Endogenous type C viruses have been detected in a wide variety of mammalian species. Multiple copies of related, but not identical, virogene sequences can be found in the DNA of these species. 2. The endogenous type C virogenes are subject to the pressures of natural selection, and closely related species possess related virogene sequences. These genes evolve as cellular entities diverging from one another in a manner which correlates well with taxonomic relatedness of the species. 3. The endogenous type C viruses of baboons and domestic cats are related, but they can be distinguished by biologic and immunologic criteria and by partial nucleic acid sequence homology. Virogene sequences in the DNA of Old World monkeys and domestic cats also show a degree of relatedness not shared by the unique sequence DNA of these species. The data suggest that progenitors of domestic cats were exogenously infected by a type C virus that also gave rise to present-day endogenous type C viruses of Old World monkeys. 4. The genomes of exogenously infectious viruses replicating in permissive host cells appear to evolve much more rapidly than endogenous virogenes which replicate as cellular genes. Laboratory strains of efficiently oncogenic type C viruses are presumed to be derived from activated endogenous viruses which have been selected for virulence and which, in certain cases, have acquired the capacity to replicate in the host's own cells. 5. The ubiquitous presence of endogenous type C viruses among vertegrates and their preservation throughout millions of years of evolution suggests that these genes express normal physiologic functions which provide a selective advantage to the species.


Reverse transcriptase associated with A-type particles from murine myeloma cells. Our findings may be summarized as follows: (a) a DNA polymerase is associated with murine type A particles, which is very similar to the reverse transcriptases found in true RNA tumor viruses; (b) the majority of the RNA isolated from A particles is of low molecular weight (probably due to degradation), byt appears to contain an amount of unique sequence information comparable to that found in other oncornavirus particles. These results suggest that A-type particles may be very similar, and perhaps related, to the two other classes (B and C) of oncornaviruses.


Properties of oncornavirus RNA-directed DNA polymerase, the RNA template, and the intracellular products formed early during infection and cell transformation. We have investigated three aspects of RNA turmor virus replication and cell transformation: (1) the properties of the purified avian and mammalian viral RNA-directed DNA polumerase, (2) some characteristics of the viral 60-70S RNA genome, 30-40S RNA subunits and intracellular viral RNA species, and (3) the interaction of the viral DNA polymerase with its RNA template early during infection and cell transformation by the murine sarcoma-leukemia virus (MSV[MLV]). Avian myeloblastosis virus (AMV) contains two forms of RNA-directed DNA polymerase, alpha, consisting of a single polypeptide of molecular weight 65,000, and alphabeta, consisting of two polypeptides of molecular weights 65,000 and 105,000. The alpha and alphabeta forms of AMV DNA polymerase both possess RNase H activity that requires free end termini on the ribopolymer and can degrade the RNA of the RNA-DNA hybrid in the 3' to 5' and 5' to 3' directions. But, alpha and alphabeta possess a different mode of exoribonuclease activity. While alphabeta RNase H is a processive exoribonuclease that degrades the polynucleotide chain to a core residue before attacking a second chain, alpha RNase H is a random exoribonuclease that releases the polynucleotide after each scission. Highly purified Moloney-MSV(MLV) DNA polymerase has both RNase H activity and the ability to read viral 60-70S RNA. These activities comigrate through five different steps of purification and are present at levels comparable to those found in purified AMV DNA polymerase. The MSV(MLV) 60-70S RNA genome and 35S RNA subunits were shown by periodate oxidationtritiated borohydride reduction to contain adenosine as the major 3'-terminal nucleoside. Poly (A) segments were isolated from viral 60-70S and 35S RNA by treatment with RNase A or RNase T1 and purified by afinity chromatography and gel electrophoresis. Viral poly(A) was shown to be present at the 3' terminus as -G(C,U)A190AOH. The similar sequence reported for poly(A) present in mammalian mRNA suggests that similar mechanisma are involved in the transcription and processing of both cellular and viral DNA sequences. Within transformed cells replicating MSV(MLV), viral 35S and 20S RNA were found in membrane-bound polyribosomes, whereas only 35S RNA was detected in free polyribosomes. The origin and function of 20S RNA is unknown. The early events during rapid infection and cell transformation of mouse 3T6 cells by the Harvey strain of MSV(MLV) were studied. By both autoradiographic analysis and molecular hybridization, viral DNA synthesis was detected in the cytoplasm by 1 hour after infection, reached a maximum at 2 hours, and subsequently decreased. Cytological chase experiments produced evidence that cytoplasmic viral DNA was transported to the nucleus. In situ hybridization experiments using radioactive viral DNA product as a probe demonstrated the rapid association of viral DNA sequences with the chromocenters of interphase nuclei and with the centromeric heterochromatin regions of some chromosomes.


Synthesis, structure and function of avian sarcoma virus-specific DNA in permissive and nonpermissive cells. We have reviewed our recent evidence for the following scheme for synthesis and integration of viral DAN after infection of permissive cells by ASV: Within the first 3 hours of infection, duplex, virus-specific DNA the length of a subunit of the viral genome (3 times 10(6) daltons) is synthesized in the cytoplasm of infected cells by a virion-associated DNA polymerase; viral DNA probably forms a covalently closed circular duplex prior to integration into host nuclear DNA. Integration and the usual consequences of viral infection can be inhibited by ethidium bromide. We have described a number of features of viral DNA prior to its integration and have indicated how these features can be exploited in the purification of viral DNA. Viral DNA has also been measured in nonpermissive (mammalian) cells in which the variable expression of viral genes is controlled by unknown mechanisms.


The genome of RNA tumor viruses: a functional requirement for a polyploid structure? This paper points out certain theoretical problems in DNA synthesis associated with antiprimer transcription and with circularization that could oblige RNA tumor viruses to rely on a polyploid genome. It is suggested that each completed act of reverse transcription may be coupled with an act of genetic recombination aimed at recovering the antiprimer information from an adjacent genome subunit in a polyploid train. A partially double-stranded DNA transcript could then be formed with sufficient terminal redundancy to permit circularization. The model provides satisfactory explanations for observed genetic interactions (particularly recombination and heterozygote formation), for inactivation data and for selective subunit transcription.


[Clinical observations of the new anti-arrhythmic drug aprindine from long-term trend analyses(author's transl)]. The therapeutic effectiveness of aprindine (Amidonal) was evaluated by trend analyses in 25 patients with predominantly severe forms of ventricular extrasystoles. The arrhythmia was stopped or improved in most instances. Severe side-effects for which the drug had to be discontinued occurred in three patients. The drug has a long half-life (about 20-30 hours). As a result, there can be cumulative dose-dependent side-effects, largely of a CNS type, disappearing after the dose has been reduced. Thirteen patients continued to be treated on an ambulatory basis, the oral maintenance dose averaging 100 mg/d, the mean follow-up period 11 1/2 months. There was no evidence of side-effects on liver, electrolytes, renal function or blood.


[The prep-trainer--thermal control in the pulp space]. The "Prep-Trainer" was developed as teaching and checking aid in order to enable students and dentists interested in continued education to check themselves. The heat development in the pulp cavity is measured at the model. Measurements made by the author showed that the safest method for maintaining constant temperatures and thus avoiding over-heating of the pulp cavity is the cooling with the air-water spray handpiece by the dental assistant.


Is rectal carcinoid argyrophilic? Application of Grimelius' silver nitrate stain in four cases. In four cases, small rectal carcinoids from patients who did not have accompanying significant clinical signs or metabolic disturbances were found to give a definite argyrophil reaction by double or triple impregnation of Grimelius' silver nitrate stain, but not by other silver methods for argyrophil or argentaffin granules, regardless of variations of cellular arrangement in ribbon-like, solid, and sclerosing patterns. The implication that the carcinoid tumor occurring in the rectum may constitute a group distinct from carcinoids arising from other sites is discussed.


[Radiotherapy of inoperable orofacial squamous epithelial carcinomas following pharmacologically induced partial synchronization]. A therapeutical method of inoperable squamous cell carcinomas based on cell kinetics has been applied since 1969 by using 5-fluorine-uracil. Impulse cytophotometric tests did not confirm the degree of partial synchronization due to 5FU that had been assumed so far on the basis of autoradiographic techniques. The results obtained in 46 patients with inoperable malignant epithelial tumors of the maxillo-facial region that were better than those obtained with radiological therapy only, can to a certain extent be contributed to 5FU. First impulse cytophotometric findings in biopsies of animal and human tumors show that Bleomycin clearly produced partial synchronization due to a reversible stop in the radiation-sensitive G2 phase.


Exocytosis of protein into the thyroid follicle lumen: an early effect of TSH. As shown in a preceding paper, only exocytotic vesicles conveying newly synthesized protein to the follicle lumen remain in the apical part of rat thyroid follicle cells following elimination of TSH secretion. In the present paper the effect of TSH on these exocytotic vesicles was investigated. TSH secretion was suppressed by administration of thyroxine for 2 days. In the electron microscope administration of TSH was seen to induce well-known signs of endocytosis, such as formation of pseudopods and colloid droplets. In addition, a previously unrecognized change was noted, namely a progressive decrease in the number of exocytotic vesicles. At 5 min after TSH the number was obviously reduced and at 20 min less than 10% of the original number of vesicles remained. Quantitative electron microscopic autoradiography after administration of [3H]leucine showed that TSH caused, concomitant with the disappearance of vesicles, a transfer of radioactivity from the apical region of the follicle cell to the periphery of the follicle lumen. The distribution of labeled protein in thyroid subcellular fractions was studied 1.5 h after administration of [14C]leucine. At 5 min after administration of TSH there was an increase of protein-bound label in the supernatant fraction, containing the luminal colloid, and a corresponding decrease of label in the particle fraction which contained most of the cell organelles, including the exocytotic vesicles. This TSH-induced redistribution of labeled proteins was more pronounced at 10 min and still more at 20 min and appeared to be dose-dependent. These observations taken together are considered to justify the conclusion that TSH induces transfer of newly synthesized protein from the follicle cells to the follicle lumen by exocytosis (i.e., emptying of specific apical vesicles). It is suggested that a causal and functional interrelation may exist between the exocytotic and endocytotic processes.


FSH stimulation of testicular androgen binding protein (ABP): comparison of ABP response and ovarian augmentation. Production of testicular androgen binding protein (ABP), ceases following hypophysectomy and can be stimulated by FSH. Within 24 h after the administration of FSH, ABP can be measured in caput epididymis supernatant and by 4 days after FSH treatment, the concentration of ABP reaches a plateau. In a 3-day assay, ABP production in immature hypophysectomized rats was stimulated by 31 mug NIH-FSH-P1 per day (0.08 U NIH-FSH-P1 per 3 days) which is comparable to the sensitivity of the ovarian weight augmentation test in hypophysectomized rats. The relative ovarian weight augmenting and ABP stimulating activities of various FSH preparations were in agreement, suggesting that the biological stimulus of the ABP response is, in fact, FSH. The ABP response to FSH could become a useful testicular bioassay for FSH. Such an assay would be more practicle if ABP could be measured by a radioimmunoassay.


Conversion of SO2 to sulfur particulate in the Los Angeles atmosphere. Gas phase and particular phase sulfur have been measured at various locations in the Los Angeles basin to determine atmospheric conversion rates and mechanisms. A new technique was developed for the measurement of particulate sulfur. From measurements of the particulate to gas phase sulfur ratio near the major stationary sources and far downstream and from estimates of travel time determined by air trajectory analysis, it is possible to estimate gas-to-particle conversion rates for sulfur. Such calculations show that automobiles presently contribute a major part of the total sulfur as measured at a receptor site such as Pasadena, while contributing only a small amount to the particulate sulfur loading. The introduction of oxidation catalyst-equipped vehicles may add significantly to the particulate sulfur at downwind receptor sites; predictions of particulate sulfur concentrations near freeways show substantial increases due to such vehicles.


Method for determining H2SO4 in automobile exhaust. A relatively simple procedure for measuring H2SO4 in auto exhaust will be presented. The system is compatible with the Federal constant volume sampler (CVS unit). The time required to get sufficient sample for titration is 15-30 min. Values on sulfates in exhaust with a catalyst car and a noncatalyst car agree well with literature data obtained by dilution tube and filtration techniques.


Operation of platinum-palladium catalysts with leaded gasoline. The effect of various fuel additives on the ability of platinum-palladium catalytic converters to remove the carbon monoxide and hydrocarbon components of automotive exhaust has been examined. Engine dynamometer studies suggest that these catalysts may be successfully used in conjunction with fuels of relatively high tetraethyllead concentrations, provided the ethylene dibromide portion of the scavenger is excluded.


Determination of sulfur trioxide in engine exhaust. Sulfur trioxide in the exhaust gas of an internal combustion engine is removed and concentrated by absorption in a solution of 80% isopropyl alcohol, which quantitatively absorbs it and inhibits the oxidation of any sulfur dioxide which may be absorbed. The absorbed sulfur trioxide (sulfuric acid) is determined by an absorption titration by using barium chloride as the titrant and thorin as the indicator. The sulfur dioxide content of the exhaust is measured continuously by means of a DuPont Model 411 ultraviolet photoanalyzer.


Measurement of vehicle particulate emissions. A constant volume sampler (CVS) compatible auto exhaust particulate sampling system has been built which samples exhaust isokinetically at constant temperature. This system yields internally consistent results and is capable of frequent and convenient operation.


Automotive sulfate emission data.

Automotive sulfate emission data. This paper discusses automotive sulfate emission results obtained by the Office of Mobile Source Air Pollution Control of EPA, General Motors, Ford, Chrysler, and Esso. This work has been directed towards obtaining sulfate emission factors for cars with and without catalyst. While the EPA and Chrysler investigations have found significant sulfate formation in noncatalyst cars, GM, Ford, and Esso have found only trace levels from noncatalyst cars. All of these investigators agree that much higher quantities of sulfate are emitted from catalyst cars. The work done to date shows pelleted catalysts to have much lower sulfate emissions over the low speed-EPA Federal Test Procedures than monolith catalysts. This is probably due to temporary storage of sulfates on the catalyst due to chemical interaction with the alumina pellets. The sulfate compounds are, to a large degree, emitted later under higher speed conditions which result in higher catalyst temperatures which decompose the alumina salt. Future work will be directed towards further elucidation of this storage mechanism as well as determining in detail how factors such as air injection rate and catalyst location affect sulfate emissions.


Catalytic converters for exhaust emission control of commercial equipment powered by internal combustion engines. The development of PTX, monolithic catalytic exhaust purifiers, is outlined, and their first use for exhaust emissions control of commercial equipment is described. The main use of PTX converters is on forklift trucks. The purification achievable with PTX-equipped fork-lift trucks under various operational conditions is discussed, and examples from the field are given. During more than ten years of operation, no adverse health effects have been reported, and PTX-equipped internal combustion engines appear safe for use in confined areas.


Estimated changes in human exposure to suspended sulfate attributable to equipping light-duty motor vehicles with oxidation catalysts. The potential environmental impact of equipping vehicles with oxidation catalysts is estimated. Three independent techniques are used to appraise community exposures. Incremental increases in suspended sulfates and sulfuric acid aerosols are evaluated in terms of the number of vehicles equipped with oxidation catalysts.


Noble metals: a toxicological appraisal of potential new environmental contaminants. The public health benefits expected by reducing known hazardous emissions from mobile sources should not be compromised by increasing levels of other potentially hazardous unregulated emissions. Catalytic converters are going to be used to meet the statutory requirements on carbon monoxide and hydrocarbon emissions from light duty motor vehicles. Platinum and palladium metals are the catalytic materials to be used in these emission control devices. Preliminary experimental evidence and analysis of the impact of these control devices on the future use and demand for platinum indicates that this metal may appear at detectable levels in the environment by the end of this decade. At the present time, platinum and palladium are not present in the public environment and represent potentially new environmental contaminants as a consequence of use of this new abatement control technology. There is relatively little information available to adequately assess the potential health hazards that may be associated with exposure to these metals and their compounds. Analysis of the environmental problems and concerns associated with possible new environmental contaminants are discussed. Limited estimates are made on community exposure by use of a meteorological dispersion model. Biodegradation potential and attention is also given to the limited toxicological information available.


Conference on health consequences of environmental controls: impact of mobile emissions controls. Conference objectives. The past and current EPA research and regulatory programs as they relate to non-regulated emissions from light-duty motor vehicles are reviewed. Provisions of Sections 202(a) and 211 of the 1970 Clean Air Act Amendments are discussed and their relationships to the nonregulated emissions issue detailed. The EPA position regarding non-regulated emissions from oxidation catalyst-equipped vehicles is discussed and related to technical papers presented in this symposium. The planned EPA research programs addressing the "catalyst" issue are reviewed.


Inhalation toxicology of automotive emissions as affected by an oxidation exhaust catalyst. Preliminary data are given on the acute inhalation toxicology of automotive emissions as affected by an oxidation exhaust catalyst. The catalyst effectively reduced CO and HC in the exhause which apparently had an effect (at least in a closed exposure system) on oxidant and NO2 levels by altering the HC/NOx ratio. There was a resultant reduction in biological effects due to the exposure. The catalyst altered the type of particulate to one which probably contained sulfuric acid as a major component. No evidence was present in these acute exposures to suggest a toxic response due to the higher sulfate emissions or possible catalyst attrition products. The effects of long-term exposure have not yet been investigated.


Preliminary studies on the toxicity and metabolism of palladium and platinum. Preliminary data are given on the LD50 of PdCl2 following different routes of exposure and on the LD50 of PtCl4 following intravenous exposure. The retention, tissue distribution, and excretion of 103Pd and 191Pt in rats was determined following oral, intravenous, intratracheal, and inhalation exposure. The highest retention for both 103Pd and 191Pt was obtained following intravenous dosing, and the lowest retention occurred after oral dosing. Following a single oral dose, almost all of the 103Pd and 191Pt was excreted in the feces due to nonabsorption, whereas after intravenous dosing, similar quantities were excreted in both the urine and feces. Tissues containing the highest concentrations of these metals were the kidney, spleen and liver. Following intravenous dosing of pregnant rats, a small amount of 103Pd and 191Pt was found in the fetuses.


Interactions of platinum metals and their complexes in biological systems. Platinum-metal oxidation catalysts are to be introduced in exhaust systems of many 1975 model-year automobiles in the U.S. to meet Clean Air Act standards. Small quantities of finely divided catalyst have been found issuing from prototype systems; platinum and palladium compounds may be found also. Although platinum exhibits a remarkable resistance to oxidation and chemical attack, it reacts chemically under some conditions producing coordination complex compounds. Palladium reacts more readily than platinum. Some platinum-metal complexes interact with biological systems as bacteriostatic, bacteriocidal, viricidal, and immunosuppressive agents. Workers chronically exposed to platinum complexes often develop asthma-like respiratory distress and skin reactions called platinosis. Platinum complexes used alone and in combination therapy with other drugs have recently emerged as effective agents in cancer chemotherapy. Understanding toxic and favorable interactions of metal species with living organisms requires basic information on quantities and chemical characteristics of complexes at trace concentrations in biological materials. Some basic chemical kinetic and thermodynamic data are presented to characterize the chemical behavior of the complex cis-[Pt(NH3)2Cl2] used therapeutically. A brief discussion of platinum at manogram levels in biological tissue is discussed.


Methods proposed to achieve air quality standards for mobile sources and technology surveillance. The methods proposed to meet the 1975 Standards of the Clean Air Act for mobile sources are alternative antiknocks, exhaust emission control devices, and alternative engine designs. Technology surveillance analysis applied to this situation is an attempt to anticipate potential public and environmental health problems from these methods, before they happen. Components of this analysis are exhaust emission characterization, environmental transport and transformation, levels of public and environmental exposure, and the influence of economics on the selection of alternative methods. The purpose of this presentation is to show trends as a result of the interaction of these different components. In no manner can these trends be interpreted explicitly as to what will really happen. Such an analysis is necessary so that public and environmental health officials have the opportunity to act on potential problems before they become manifest.


Motor neurone disease: an immunological study. The state of the immune system, both humoral and cell-mediated, was evaluated in motor neurone disease (MND) patients. The data obtained for the MND patients were confronted with normal controls and a group of patients affected by nervous diseases not involving the immune system. Some differences were observed between MND patients and normal subjects, namely: increase of WBC and gamma-globulin in MND patients. However, such differences were not observed between MND patients and pathological controls, and therefore are probably due to a higher frequency of infectious complications in MND patients in respect to normal controls. The capacity of the immune system to respond to an adequate stimulus was normal, and no precipitating anti-CNS antibodies were detected in MND sera. Furthermore, no sex-linked differences were observed and the CSF abnormalities observed in 2 out of 16 MND patients were probably reflecting only destruction of CNS cells. The data are discussed in view of the possible pathogenetic mechanisms of MND.


Cerebrospinal fluid protein patterns in spasmodic torticollis. In order to investigate the value of CSF-protein analyses in spasmodic torticollis CSF from six patients with probable organic and two patients with probable psychogenic torticollis was examined by isoelectric focusing and electrophoresis. In all the patients with organic torticollis two pathological CSF-protein fractions were found in the alkaline region on electrofocusing and in four cases aberrant fractions occurred also in the acidic pH range. An increasing number of abnormal fractions were noted during at least the first year after onset of symptoms. Lithium treatment of three patients resulted in a striking decrease of torticollis as well as of the number of abnormal CSF-protein fractions. During placebo treatment of two cases, torticollis and the pathological CSF-proteins recurred. Some observations, including a few previous autopsy findings, might indicate that an encephalitogenic agent is involved in the pathogenesis of organic torticollis. In the patients with psychogenic torticollis the CSF-protein pattern was normal. This investigation supports a recent suggestion that organic and psychogenic torticollis might be distinguished by electrofocusing of the CSF-protein.


A combination of sister chromatid differential staining and giemsa banding. We report a procedure for combining sister chromatid differential staining and G banding in the same metaphase plate. Mammalian cells in culture are grown in medium containing 5-bromodeoxyuridine for two cell cycles, and conventional air-dried preparations are made. The slides are treated with a trypsin or a urea solution the same way as for regular G banding. This method is simple and fast and provides additional information for cytogeneticists.


[Relation of TRH test to thyroidal suppression test by triiodothyronine in patients with hyperthyroidism under treatment with antithyroid drugs (author's transl)]. The relation of the TRH test to the T3 suppression test was investigated in 43 patients with hyperthyroidism receiving antithyroid drugs for 6 to 27 months (average 14 months). TRH tests were performed by measuring serum TSH levels before and 15, 30, 45, 60, 90 and 120 minutes after intravenous injection of 500 mug of synthetic TRH. Serum TSH was measured by a double antibody radioimmunoassay. Two weeks later, the T3 suppression test was performed by measuring the 24-hr thyroidal uptake of radioiodine after daily administrations of 75 mug of triiodothyronine for 8 days. All patients under study were in euthyroid state, estimated by serum T3-RSU, T4, T3 and FT4I. The value for 24-h uptake after T3 administration was less than 20% in 18 cases, out of which the response to TRH was normal or exaggerated in 15 cases and was absent or impaired in 3 cases. On the other hand, out of 25 cases with the value of 24-h uptake more than 20%, the response to TRH was absent or impaired in 18 cases and was normal or exaggerated in 7 cases. The results of the TRH test and the T3 suppression test were correlated in 33 out of 43 cases. The responsiveness to TRH and the suppressibility with T3 was dissociated in 10 cases, suggesting the TRH test could not replace the T3 suppression test.


[Studies on the RNA synthesized in the ovary of immature rats after HCG administration (author's transl)]. Although many researchers have reported that RNA synthesis in the ovary is enhanced by gonadotropin treatment, there are only a few papers concerning the character of newly synthesized RNA after gonadotropin treatment. In this paper, the RNA synthesized in the ovary of immature rats after HCG treatment was qualitatively studied. Immature female Sprague-Dawley rats were administered with 0.3 mc per rat of 3H-uridine at a certain time interval after injection of HCG (10 iu/rat) and the ovaries were subsequently isolated after 15, 30 or 60 minutes. RNA was extracted from the homogenate of the ovaries according to the hot phenol method after Scherrer and Darnell. The 3H-RNA thus extracted was treated with electrophoretically purified DNase to break down and remove DNA that mingled with it. The RNA solution ultimately obtained was analysed on a 3-20% sucrose gradient. The different fractions thus separated were then subjected to measurement of radioactivity and optical density at 260 mmug. The RNA extracted from the ovary of immature untreated rat labeled with 3H-uridine for 15 minutes showed a flat pattern of radioactivity from the top to the bottom fractions with low radioactivity. Otherwise, when labeled for one hour, the RNA showed a pattern of radioactivity like those of optical density at 260 mumu with peaks of r-RNAs and t-RNA. When the ovary was pulse-labeled with 3H-uridine for 15 minutes starting 2 hours after injection of HCG, the RNA with a large S value was synthesized and the pattern of variation in radioactivity was that of rising near the bottom fraction and declining with access to the top fraction. The results obtained by labeling for 15 minutes starting 40 hours after PMS administration were similar to those obtained in immature untreated rats. The patterns of radioactivity in RNA obtained by the labeling for 15 minutes starting 2 hours after HCG and 42 hours after PMS were similar to those starting 2 hours after only HCG injection. The patterns of radioactivity became similar to those of optical density at 260 mmu, when the ovaries were labeled for 30 or 60 minutes. From these results, it was suggested that the newly synthesized RNA 2 hours after HCG was constructed from m-RMA with rapid turn over and precursors of r-RNAs and t-RNA. This RNA synthesis was blocked by pretreatment with actinomycin but not by cycloheximide. From these results, it was suggested that enhancement in RNA polymerase activity or change in template capacity of DNA which would have an effect on RNA synthesis was not based on newly synthesized protein.


Carbohydrate tolerance in autoimmune thyroiditis. Glucose tolerance tests were carried out on fifty patients with autoimmune thyroiditis of varying duration and severity. Two were floridly diabetic, and a further six showed diabetic abnormalities of glucose tolerance, giving an over-all incidence of 16 per cent. Diabetics were significantly older than nondiabetics, but the two groups did not differ in terms of duration of thyroid disease or frequency of associated disease of probable autoimmune origin. The prevalence of diabetes in patients with autoimmune thyroiditis appears to be the same as that in the population generally.


Release of proteolytic enzymes in bile-induced pancreatitis in dogs. Pancreatitis was induced by injection of autologous bile into the main pancreatic duct of dogs. An initial fall in blood pressure was accompanied by appearance of large quantities of active trypsin, chymotrypsin, and elastase in pancreatic exudate with full saturation of protease inhibitors. The enzymes soon appeared in ascitic fluid and lymph, but only in the form of complexes with alpha1-antitrypsin, and alpha2-macroglobulin. No such complexes were detected in venous blood indicating short half-life in the circulation. These studies confirm the release of pancreatic enzymes during bile-induced pancreatitis, and quantify an important protective role for plasma protease inhibitors in this situation.


Studies on the toxicity of lindane on Colisa fasciatus (part I: TLm measurements and histopathological changes in certain tissues). 1. In the present investigation static bioassay studies and histopathological changes in some tissues (gills, liver and kidney) induced by Lindane 20% E.C. in the fish Colisa fasciatus have been taken under consideration. 2. With the help of bioassay studies, the TLm were noted. The TLm values were 0.87, 0.78, 0.68 and 0.64 mg/l for the temperature range 17 degrees C to 20 degrees C and 0.60, 0.56, 0.46 and 0.41 mg/l for the temperature range 32 degrees C to 35 degrees C at 12, 24, 48 and 96 h respectively. 3. The statistical analysis of different factors such as concentrations, time and temperature range based on the observed data gave the following results: a) At both temperature ranges, survival rates at different concentrations and time are significant at 1% and 5% level. b) The difference in survival rates at different temperature range are insignificant both at 1% and 5pected survival numbers. The standard errors were also calculated for these two lines which gave the insignificant results and there was no much change in the shape of experimental and expected lines. 4. The histopathological changes in the tissues (gills, liver and kidney) shows the following results: a) In the case of the gills there was a loss of different types of cells i.e. respiratory cells and blood cells. Blood vessels were atrophied and the outer membrane of gill lamellae was ruptured. The erosion of the tips of the gill filaments were also observed. b) In the case of liver the outer membrane was ruptured. Some hepatic cells were vacuolated and some completely degenerated. The central areas were much affected. Due to the toxic effect, large splitting of tissues were found inside the liver. c) In the case of kidney the outer epithelium, the parenchymatous cells of renal tubules, cuboidal cells of uriniferous tubules were much affected. Due to the degeneration of the cells large spaces were formed inside the tissue. Much affection was observed in the central areas.


[Cytochemistry of leukocytes in children. II. Changes in cytochemical reactions of neutrophil granulocytes in the course of leukoses]. A report is presented on the changes of cytochemical reactions of neutrophilic granulocytes in children during acute leukaemia. Various hydrolases revealed a significant increase of activity during the acute stage of disease. The enzyme activity begins to decrease in remission, but regains its normal value only in the activity of alkaline granulocytic phosphatase. The glycogen content of granulocytes is reduced at the beginning of the disease and during the recidive as well as in the granulocytes of children affected with chronic myeloid leukaemia. The enhanced enzyme activity of granulocytes refers to the fact that the defence function of cells in acute leukaemias is not completely cancelled. Only a decreased glycogen content of cells refers to a disturbance in the metabolism of granulocytes.


Gamma-D paraproteinemia - a report of 16 cases. Paraproteins of the gammaD class are very rare, being found in only 0.3% to 3.0% of the cases of all paraproteinemias. They are difficult to detect and therefore sometimes not found or mis-diagnosed. In the 6500 cases of paraproteinemia we have diagnosed over the last eight years, 16 of them have been gammaD. Consistent with the reports of other gammaD paraproteinemias most of these 16 cases were of type L (13), most patients were male (11), the average age (55), was younger than that of patients suffering from other paraproteinemias, and in all cases the disease followed a rapidly malignant course.


[Studies on changes in lymphocyte HL-A antigens during the course of malignant diseases]. The presence of HL-A antigens 1, 2, 5, 7, 8 and 12 on the lymphocytes of 26 patients with blood diseases and malignant tumours was examined by means of the two-step microcytotoxicity test. The studies carried out several times in the course of the disease and with 4 to 5 sera of the same specificity. Two types of the serological modifications were found: 1. Transient loss of HL-A antigens in 6 patients, 2. Evidence of the polyreactivity of lymphocytes in 3 patients. The polyreactivity was later changed to the loss of HL-A antigens. In one patient, the destruction of lymphocytes in the course of the testing was proved. The importance of the results is discussed. The additional serological, morphological and clinical investigations seems to be necessary.


[Genetic relationships between the HL-A system, the blood group systems (Rh, ABO) and the immune reactivity]. The authors examined in immunized healthy persons the correlations between the ability of immune response, the value of their different immunological parameters, and the HL-A blood-group antigens by computer analysis. Immune reactivity showed mosaic-like correlation against the HL-A system. The most definite negative correlation was noticed between the HL-A 3 and 7 antigens and the cytotoxic activity of lymphocytes. Remarkable and definite correlation was found between the Rh system and immune reactivity. The level of the natural antibodies, the immunoglobulins and the functions of lymphocytes were generally decreased in males in comparison to females.


Studies on delayed hypersensitivity of patients with Hodgkin's disease during remission. The studies of skin sensitivity, the reactivity to intradermally transferred lymphocytes and lymphocytic transformation in vitro showed that delayed hypersensitivity had been slightly impaired in patients with Hodgkin's disease during long time remission. The regression of the disease favoured the recovery of the lymphocytic functions and the delayed type immune reactions associated with them. These data suggest that the enhancement of the delayed hypersensitivity of the tumour defence respectively could promote the treatment or prevention of the disease.


[Studies on the production of anti-human-lymphocyte serum (ALS) in bulls]. The applicability of bulls as productive animals was considered for the preparation of anti-humans ALS. The course of immunologic response was studied by lymphoagglutination, lymphocytotoxicity, rosette inhibition, hemagglutination tests and by precipitin formation in two experimental groups immunized by different amounts of lymphocytes from peripheral blood of normal donors. The animals were found to respond well already after the second application of very small amounts of antigen (on day 0-4 times 10(7), on day 21-2 times 10(8) lymphocytes). They showed lymphoagglutination titre 1 : 512-2000, lymphocytotoxic titre being higher than 1 : 4000 and the rosette inhibition test gave a minimum titre of 1 : 65000. On the other hand, further application of a high amount of antigen (2 times 10(9), or 4 times 10(9) lymphocytes) did not lead to further increase in the titre; on the contrary - hyperimmunization resulted in a lower titre in the case of the rosette inhibition test, which is known to correlate best with the in vivo immunosuppressive activity. The hemagglutinin titre was also acceptable under the above conditions and the formation of undersirable precipitins against human serum proteins was negligible. Good response reached by a simple and economical immunization scheme speaks for the suitability of bulls for the production of ALS.


[Use of the immune tolerance induction in the production of an antilymphocyte serum (ALS) free of antibodies against serum proteins]. The possibility has been investigated of a direct gain of ALS free of undesirable antibodies against serum proteins by inducing immunologic tolerance in productive animals (pigs). Preliminary experiments made with tolerogenic amounts of 10 and 50 ml of sera and with immunization by the serum alone proved applicability of this method. Electrophoresis showed antibodies against 6 to 7 and 2 to 3 fractions in animals tolerated with 10 and 50 ml respectively, compared to 18 to 20 fractions in the control group, which was not tolerated. This has been confirmed when preparing ALS in practice, where the toleration was carried out with 25 ml of serum or with the same amount of serum with the addition of hemoglobin and immunization by lymphocytes isolated from peripheral blood. Final ALS of untolerated animals contained antibodies against 7 to 8 fractions, whereas that of experimental group tolerated with serum and Hb was free of antibodies against serum protein, hemoglobin included. ALS of the group tolerated with normal serum contained only antibodies against hemoglobin. In vitro tests (i.e. lymphoagglutination t., lymphocytotoxicity t., rosette inhibition t.) proved that by inducing tolerance towards serum protein the activity of ALS was in no was affected. According to the results this method can be employed not only for the preparation of ALS, but also for other purposes, such as preparation of monovalent antisera for immunoelectrophoresis.


Proliferative activity of erythrone and intramarrow hemolysis in iron deficiency anemias. 3H-thymidine incorporation into normoblasts, proliferation rate of erythroid precursors and degree of intramarrow hemolysis have been studied in vitro on the bone marrow. The normal proliferation rate of normoblasts is 26 +/- 2% i.e. during 24 hours about a quarter of dividable elements of erythropoiesis is renewed. Acute blood loss increases the proliferation rate up to 57 +/- 9% but the value of 3H-thymidine incorporation into cells is not changed as compared to normal. In chronic blood loss both 3H-thymidine incorporation into dividing erythroid precursors at different stages of maturity and the rate of erythroid production are 2 to 3 times lower than normal. In healthy persons the degree of intramarrow hemolysis is 7 +/- 2% of erythroid precursors incubated for 24 hours. In iron deficiency anemia intramarrow destruction sharply increases, presenting at an average 30% of incubated nucleated elements of erythropoiesis. A type of chronic iron deficiency, which is not associated with blood loss, is described. In this type of anemia the proliferation rate of normoblasts and the degree of intramarrow hemolysis do not differ from normal values.


[The Breddin test in various forms of idiopathic thrombocytopenic purpura]. In 100 healthy persons and 38 patients with idiopathic thrombocytopenic purpura (ITP) the morphology of thrombocytes was examined according to BREDDIN. The survival time of the marked thrombocytes and their place of sequestration was determined in the patients and in 17 voluntary persons. A correlation was found to exist between the morphology of thrombocytes and those four forms of ITP which could be limited by means of radiometric methods. Moreover, a certain degree of dependence was found between the thrombocyte sequestration place and the presence of antibodies.


[The determination of platelet factor 3 using hirudin]. The influence of blood platelets on the recalcification time under hirudin was investigated. Contrary to the investigations of whole-blood which reveal a pathological prolongation of the hirudin tolerance test only at platelet numbers under 30,000/mug, a change of the recalcification time under hirudin could also be found in the plasma at higher platelet numbers. The recalcification time increased inversely proportionally with falling platelet number. The shortening of time in platelets rich plasma attributed to the activity of platelet factor 3. Differences in the examinations of whole-blood may be attributed to an erythrocyte activity similar to factor 3. The application of hirudin for determining platelet factor 3 is recommended as a sensible method easily to be performed in practice.


[Liberation of anti-heparin activity during platelet aggregation in patients with blastic leukosis and blastic crisis of chronic myelosis]. The activity of platelet factor 4 (FP4) was examined in 37 patients affected with blastic leukaemia, in 16 patients with blastic crisis in chronic myeloid leukaemia and in 2 patients with Willebrand's syndrome. The real activity of FP4 determined by modifying the method according to NIEWIAROWSKI after a complete lysis of granular membrances by means of triton X-100 was found to be lowered in 9 patients affected with blastic leukaemia, in 5 patients with blastic crisis and in two patients with Willebrand's syndrome. Presuming that a maximal FP4 release of the irreversible platelet aggregation must be obtained, which corresponds to the real activity, the author has examined the apparent activity of FP4 released from the aggregated platelets with the help of her own method. In this way the quality of the release reaction from the platelets can indirectly be characterized with their extremely important role for haemostasis. Whereas in exacerbated myeloid leukaemia and Willebrand's syndrome the apparent activity will correspond to the real one, there is a severe specific disturbance of the platelet release response in blastic leukaemia. The platelet aggregation is incomplete caused by the derailment of the energy metabolism and the disturbance of the adenine nucleotides, thus causing an apparent as well as a real FP4 deficiency which can be brought into the same line with pathogenesis of thrombopathy in blastic leukaemia.


Effect of testosterone-estradiol-binding globulin in the enzymic oxidoreduction of 17-oxygenated c19 steroids. The effect of both testosterone-estradiol-binding globulin (TeBG) and albumin on enzymic oxidoreduction of four 17-oxygenated C19 steroids by bacterial 17beta-hydroxysteroid:NAD oxidoreductase from Pseudomonas testosteroni was investigated. The decreased yields of products under presence of TeBG were found in both directions of reversible enzymic reaction. This finding was unexpected in the case of enzymic reduction in which the opposite effect could be assumed with respect of high affinity of the product but not the substrate to TeBG. Kinetically, the competition between enzyme and binding protein for the substrate occurs at enzymic oxidation, whereas the mechanism resembling non-competitive inhibition operates in the enzymic reduction.


[The primary structure of a monoclonic immunoglobulin-L-chain of subgroup IV of the kappa type (Bence-Jones protein Len.)]. The complete amino acid sequence of Bence-Jones protein Len. was established by sequential analysis of tryptic and chymotryptic peptides. The result of these experiments and the comparative sequence analysis with the other 17 completely determined kappa-proteins is incompatible with the serological typisation of protein Len. as a member of subgroup II: There are 18 positions in protein Len. than cannot be associated with any one of the subgroups kappaI, kappaII or kappaIII. Also the average amino acid exchange rate between protein Len. and these subgroups is in the same range as the average amino acid exchange rate between these subgroups. Therefore Bence-Jones protein Len. is the first completely determined representative of a new IV. kappa-type subgroup. The variability of immunoglobulins follows a structural principle in which the single point mutations responsible for the variability are linked. The present paper contains the exact analysis of the linked point mutations within the so far best-investigated subgroup, kappaI (12 completely sequenced proteins). These linked exchanges allow the arrangement of the kappaI proteins in 4 subgroups and their further subdivision. The regularity of this amino acid sequence pattern can only be explained by an evolutionary origin of antibody variability. On the basis of this evolutionary mechanism the relationship of immunoglobulins can be depicted in a phylogenetic tree. Such a tree was therefore constructed for the 18 completely determined Bence-Jones proteins of kappa-type, for the first time taking into account Bence-Jones protein Len. Its topology is in complete agreement with the results of the comparative sequence analysis.


[Isolation of beta2-microglobulin from human colostrum (author's transl)]. The beta2-microglobulin from human colostrum was purified by a combination of ordinary protein-chemical techniques: gel filtration, ion-exchange chromatography and zone electrophoresis. The procedure is organized in such a way that the simultaneous isolation of many other milk proteins is possible. The beta2-microglobulin obtained from colostrum cannot be distinguished by physical-chemical or immunological means from the beta2-microblobulin isolated from the urine of patients with kidney-tubule diseases. At the beginning of lactation, human milk contains significantly more than 10 mg/-100 ml beta2-microglobulin, but the concentration drops within two or three days to 15-30% of the original amount.


Human polymorphonuclear leucocyte migration inhibitory factor. Evidence for antigen dependency. Sudies were performed on human polymorphonuclear leucocyte migration inhibitory factor (PMN-MIF) to determine its antigen dependence. PMN-MIF produced by lymphocytes in response to purified protein derivative or coccidiodin was measured in an agarose gel system with buffy coat leucocytes as indicator cells. PMN-MIF activity contained in the lymphocyte supernatants uniformly disappeared when the supernatants were diluted 1:50 with medium; the inhibitory activity was only restored when the diluted supernatants were reconstituted with specific antigen. PMN-MIF isolated by polyacrylamide gel electrophoresis showed the same properties as PMN-MIF present in whole supernatants. This factor consistently migrated in the albumin region on gel electrophoresis. These data indicate that human PMN-MIF is antigen-dependent.


Cellular co-operation by mouse lymphocytes to chemically defined antigens. When populations of mouse spleen cells were treated with a highly radioactive 125I-labelled haptenic conjugate (125I-labelled PDG-N) and subsequently injected into X-irradiated, immunized (N-10-C) syngeneic BALB/c mice, it was found that after autoradiography with 125I-labelled N and C haptenic conjugates, the numbers of ABC to both the N and C haptens decreased. The number of ABC found were significantly lower than those found in mice that had been reconstituted with cells "suicided" with another 125I-labelled antigen (125I-labelled BSA) and subsequently immunized with N-10-C.