Hepatic dearterialization for nonrespectable primary and secondary tumors of the liver. Seventeen patients with primary or secondary liver tumors were treated by dearterialization of the liver and intraportal infusion of cytotoxic drugs. In 14 cases, ligation of the main (10), right and left (1), right and middle (1), and right (2) hepatic arteries was performed. In three in whom hepatic artery ligation was considered to be contraindicated due to occlusion of the portal vein, temporary occlusion of the hepatic artery was repeated postoperatively. Only one patient died from the cause related to the ligation. Out of 14 patients who survived for more than one month ten showed an apparent regression of tumors radiologically. The mean survival period of the patients excluding those who died from other unrelated causes was 28 weeks. That of the control patients who received only chemotherapy through the hepatic artery was 13 weeks. The results were not always satisfactory, but dearterialization of the liver can be a palliative treatment for nonresectable liver tumors with a forceful combined chemotherapy.


Radioimmunoassay of neocarzinostatin, an antitumor protein. Antibodies directed toward the antitumor protein neocarzinostatin (NCS) have been produced in a rabbit by immunization with a highly purified NCS preparation. The antiserum was monospecific and reversed the antibacterial activity of NCS against Sarcina lutea. It cross-reacted with chemically modified derivatives of NCS and mitomalcin but failed to cross-react with macromomycin. A radioimmunoassay procedure has been developed utilizing the antiserum and a biologically active 125I-labeled derivative of NCS. The lower limit of detection by this radioimmunoassay, which involves a double antibody technique for the separation of antibody-bound and free antigen, was 1 X 10(-13) mole. The sensitivity of the assay is such that serum levels of NCS can be determined accurately after administration of the drug to rats at a single dose of 2 mg/kg. Since NCS is now undergoing clinical trial, the radioimmunoassay of the drug will be a valuable tool in clinical pharmacological studies.


alpha-Fetoprotein in toxic liver injury. The temporal sequence of alpha-fetoprotein appearance in serum was determined in both necrogenic and nonnecrogenic liver injury. Ethionine, thioacetamide, and CCl4 were used to intoxicate male and female rats for evaluating serum enzyme levels, mitotic indices, and morphological reflections of impairment. Thioacetamide- and CCl4-induced cell death preceded the mitotic wave in residual hepatocytes, and, in the case of both agents, this intoxicant-mediated necrosis preceded the emergence of alpha-fetoprotein. Yet, although there was no evidence of either cell destruction or significant mitotic activity in ethionine-poisoned animals, serum alpha-fetoprotein levels progressively increased. Thus the temporal sequence of alpha-fetoprotein synthesis and/or release and cellular reorganization for regeneration suggests that reappearance of the protein macro-molecule is an expression of the altered phenotype observed during the "step-down" phase of liver regeneration.


Genetic control of interindividual variations in the inducibility of aryl hydrocarbon hydroxylase in cultured human lymphocytes. Interindividual and intraindividual variations in aryl hydrocarbon hydroxylase (AHH) induction by 3-methylcholanthrene were studied in cultured lymphocytes from normal adult volunteers. Using eight pairs of monozygotic and eight pairs of dizygotic twins, we examined to what extent these variations are controlled by heritable factors and whether AHH inducibility correlations in an individual with the plasma half-lives of three drugs. Substantial overestimation of the induction ratio (fold inducibility) may occur if the nonlinearity of the assay standard curve is not considered. Fold inducibility remains relatively constant for an individual, but large intraindividual variations occur in absolute "control" and "induced" AHH activities. Fetal calf serum may contain inducers of AHH activity that vary with the particular lot of serum, thereby rendering the apparent induction ratio an imprecise indicator of genetic susceptibility to induction by 3-methylcholanthrene. The index of heritability for AHH fold inducibility in twins studied with different lots of fetal calf serum (0.80) or with a single lot of fetal calf serum (0.77) suggests nonetheless that genetic rather than environmental factors are mainly responsible for interindividual variations in AHH inducibility by 3-methylcholanthrene in human lymphocytes. In these twins a significant but poor correlation (r=-0.551; 0.03 less than p less than 0.05) occurs between AHH inducibility in culture and the plasma antipyring half-life, but not between AHH inducibility and phenylbutazone or bishdroxycoumarin half-lives.


Absence of thymus-derived lymphocyte markers in myelogenous leukemia (Ph1+) cell line K-562. The myelogenous leukemia cell line K-562 with a Ph1+chromosome, derived from a patient with chronic myelogenous leukemia in terminal blastic crisis, is not a bone marrow-derived lymphoblastic cell line, because the cells neither produce immunoglobulins nor possess complement receptors. Since it has been suspected that blasts found in some patients with chronic myelogenous leukemia in blastic crisis might be thymus-derived cells, we have studied several parameters to demonstrate that K-562 cells are not thymus-derived lymphoblasts. The results of this study show: (a) no cross-reactivity of antisera to K-562 cells with normal human thymocytes; (b) lack of cytotoxicity of a specific horse anti-human thymocyte globulin for K-562 cells; (c) failure of the treatment of K-562 cells with bovine thymosin to induce antigenic determinant and erythrocyte rosette receptors on K-562 cells; (d) presence of receptors for the Fc portion of immunoglobulin G; (e) absence of terminal deoxynucleotidyl transferase; and (f) cytotoxicity of monkey antiserum to K-562 cells for malignant thymus-derived cells (Molt-4). However, absorption with Molt-4 cells abolished the cross-reactivity with Molt-4 cells, whereas 60% of the antibody to K-562 cells remained in the immune serum. Studies of DNA polymerase activities revealed that K-562 cells have levels of polymerase alpha and beta, like other proliferating cells, and an RNA-dependent DNA polymerase activity, presumably representing polymerase gamma.


Evidence for a particle-associated RNA-directed DNA polymerase in rabbit placental and uterine tissues. A RNA-directed DNA polymerase associated with particles that band at a density characteristic of type C RNA viruses was found in normal rabbit placental and uterine tissues taken during the early stages of gestation. That the rabbit RNA-directed DNA polymerase is distinct from the known cellular DNA polymerases and similar to the RNA-directed DNA polymerase of mammalian type C RNA viruses is shown by column chromatographic characteristics, template primer preferences, molecular weight determination, and an absolute requirement for the divalent cations.


Presence of a high-molecular-weight RNA and RNA-directed DNA polymerase in rabbit hereditary lymphosarcoma. Rabbit lymphosarcoma tissues contain 70 S RNA and RNA-directed DNA polymerase encapsulated in particulate components that band in the density region of type C RNA viruses. RNA-directed DNA polymerase associated with the particles could be distinguished from cellular DNA polymerases by salt elution from phosphocellulose. The enzyme preferred the template primers poly(rA)-(dT)12-18 and poly(rC)-(dG)12-18 over other synthetic template primers and also utilized viral 70 S RNA as template; these properties are not observed with the known cellular DNA polymerases.


Two-stage control of cell proliferation induced in rat liver by alpha-hexachlorocyclohexane. Determinants of the timing of DNA synthesis in rat liver were studied, using alpha-hexachlorocyclohexane as a tool for stimulation of cell proliferation. One determinant is the time of alpha-hexachlorocyclohexane administration. The increase in DNA synthesis starts after a lag phase (prereplicative phase) of minimally 20 hr. Use of animals adapted to a controlled feeding and lighting schedule revealed a second determinant provided by food consumption. Initiation of DNA synthesis is suppressed by fasting or protein deprivation and occurs 5 to 8 hr after readministration of a protein-containing diet. The light-dark rhythm has no direct influence on the timing of DNA synthesis. Stimulation of hepatic DNA synthesis, therefore appears to require two different sequential signals. The first is provided by alpha-hexachlorocyclohexane, and the second is provided by protein intake. In the absence of the second signal, committed cells are arrested at a critical point of the prereplicative phase and accumulate. Protein intake permits release from the block, and the accumulated cells enter the S period almost synchronously after completion of the remaining 5 to 8 hr of the prereplicative phase. These observations provide a means of synchronizing, in the living animal, a proliferating population of hepatocytes. In addition, they offer an explanation for the diurnal rhythmicity in the rate of hepatic cell proliferation.


Reduction of N-nitrosodiethylamine carcinogenesis in rats by lipotrope or amino acid supplementation of a marginally deficient diet. In studies in this and other laboratories, induction of hepatocardinoma by several different chemical carcinogens was enhanced in rats fed diets deficient in lipotropes (choline, methionine, folic acid), amino acids, and niacin, and high in fat. In some cases, specific supplementation with lipotropes blocked carcinogenesis. In studies reported here, specific supplementation of a marginally deficient diet that enhanced carcinogenesis in rats, with the amino acids or lipotropes in which it was deficient, significantly decreased induction of hepatocarcinoma by N-nitrosodiethylamine. Niacin supplementation decreased hepatocarcinoma incidence only slight; the addition of beef fat to an adequate diet did not enhance tumor induction. Rats fed the amino acid- or lipotrope-supplemented diets had an increased incidence of hepatic hemangioendothelial sarcomas, compared to deficient rats or to rats fed the adequate control diet. Methionine was contained in both the amino acid and the lipotrope supplement and probably was responsible for reducing hepatocarcinoma incidence. Methionine has been found to have an anticarcinogenic effect in other studies and also to block the depletion of hepatic folate stores that is induced by N-nitrosodiethylamine. Interactions between carcinogens, S-adenosylmethionine, and folate may be significant in hepatic or other tissue carcinogenesis. One of more hepatic microsomal oxidases were depressed in rats fed any of the high-fat diets but were not correlated with tumor incidence.


Common tumor rejection antigens in methylcholanthrene-induced squamous cell carcinomas of mice detected by tumor protection and a radioisotopic footpad assay. Seven transplantable lines of squamous cell carcinoma of the skin initiated in BALB/c mice by skin painting with methycholanthrene were systematically tested for cross-reactivity of their tumor rejection antigens in a 7 X 7 matrix. As determined by decreased tumor frequency after tumor cell challenge, each line was immunogenic against and/or immunosensitive to at least one and usually more than one of the other lines. A radioisotopic footpad assay for delayed hypersensitivity against viable tumor cells confirmed the cross-reactivity shown by tumor rejection. More than two antigens appeared to be present in the lines. Tests for C-type viruses were positive in all tumors; those for polyoma virus were negative. Whether the uniform presence of C-type viruses can account for the number and variety of antigens found, or whether the tumor rejection antigens are independent of virus expression, remains an open question. The finding of cross-reacting tumor rejection antigens in methylcholanthrene-induced squamous cell carcinomas encourages prospects for the development of more broadly applicable immunodiagnostic and immunotherapeutic reagents.


Bleomycin (NSC-125066) followed by cyclophosphamide (NSC-26271), vincristine (NSC-67574), methotrexate (NSC-740), and 5-fllorouracil (NSC-19893) for non-oat cell bronchogenic carcinoma. The in vivo observation that bleomycin may be used as a synchronizing agent provides the basis for testing 4 days of continuous bleomycin infusion followed by 5 days of intensive chemotherapy with cyclophosphamide, vincristine, methotrexate, and 5-fluorouracil. Thirty-eight patients with extensive non-oat cell bronchogenic carcinoma (adenocarcinoma[17 patients], squamous cell carcinoma[14 patients], and poorly differentiated carcinoma [seven patients]) were registered for chemotherapy. There were 11 patients with 50% regression of all measurable lesions and four with improved but poorly measurable radiographic lesions, providing a crude response rate of 39% (15 of 38 patients). An overall survival median of 19 weeks compares favorably with Veterans' Administration Lung Cancer Study Group control data, but was not substantially better than our own historical controls (P = 0.15). The median survival for responders was 36 weeks compared to 16 weeks for historical controls (P = 0.001) and 12 weeks for nonresponders (P less than 0.001).


Phase I clinical trial of combined therapy with vinblastine (NSC-49842), bleomycin (NSC-125066), and cisdichlorodiammineplatinum(II)( NSC-119875). Seventeen patients with various histologic types of incurable malignant disease were treated with a combination of vinblastine, bleomycin, and cis-dichlorodiammineplatinum(II). Creatinine and blood urea nitrogen elevations were noted but were not of a severe degree. White blood cell and platelet count depressions were seen and appeared to be cumulative, though not life-threatening. Tinnitus and high-frequency hearing loss were noted. Tumor regression was seen in one patient with adenocarcinoma of the lung and in one patient with a testicular tumor. This appears to be a manageable drug combination with frequent monitoring of renal, hematopoietic, pulmonary, and auditory function. A phase II study establishing the therapeutic efficacy of this combination in advanced testicular neoplasms now appears to be indicated.


Perivascular regions of the rat neural lobe. Neural lobes and portions of occipital cortex from rats were examined electron microscopically following fixation in 4% tannic acid in 2.5% glutaraldehyde. The procedure allowed a clear demonstration of the perivascular space and intercellular spaces in both tissues. The perivascular spaces in the neurosecretory tissue was far more extensive than in the neural tissue and the role of this region in relation to the process of neurosecretion is discussed.


Anatomy of the ocellar interneurons of acridid grasshoppers. I. The large interneurons. The anatomy of the large ocellar interneurons in the brain of five species of acridid grasshoppers of two different subfamilies (Schistocerca vaga, S. gregaria, Gastrimargus africanus, Trimerotropis pallidipennis, and Arphia conspersa) was revealed by cobalt-filling of the three ocellar nerves and subsequent reconstructions from silver-intensified (Timm's method) serial sections. Conflicts in the literature are reviewed (Tables 1, 2) and differences in the number of cells, anatomical descriptions of these cells, and nomenclature are resolved by demonstration of an identical number of large ocellar identical number of large ocellar interneurons in all five species examined (Fig. 1). There are 17 large 1st-order ocellar interneurons (Figs. 2, 3). Each of the three ocellar nerves contains the axons of seven large interneurons; four of these interneurons have axons in two ocellar nerves. The anatomy of three pairs of 2nd-order ocellar interneurons (with branches in the ocellar tracts within the brain and axons in the circum-esophageal connectives) is reconsidered in light of recent conflicts in the literature. Previous accounts by Williams (1975) of interneurons O2, O3, and PI(2):5 are corroborated and new details added (Fig. 7) by the use of a cobalt method that appears to stain these 2nd-order interneurons transsynaptically (Fig. 6).


Anatomy of the ocellar interneurons of acridid grasshoppers. II. The small interneurons. The anatomy of the small ocellar interneurons in the brain of the acridid grasshopper Schistocerca vaga was revealed by cobalt-filling the three ocellar nerves and subsequent reconstructions from silver-intensified (Timm's method) serial sections. In total, 61 small ocellar interneurons were repeatedly identified with arborizations in many areas of the brain and optic lobe, including in particular the posterior neuropil, ocellar tracts, protocerebral bridge, lobula, ventral bridge and tritocerebral crotch, calyces, and antenno-glomerular tracts. Each ocellar nerve contains the axons of small cells that arborize in the other two ocellar tracts; these tracts are sites of ocellar integration. Direct interactions between the ocelli and compound eyes are suggested by the projections of small ocellar interneurons into the proximal lobula. Small cell arborizations from all three ocelli are distributed actoss much of the protocerebral bridge, implying a role for the bridge as an ocellar neuropil within the brain. Four of the small interneurons could be seen in whole-mount preparations and are demonstrated to be identical in five species of acridid grasshoppers of two different subfamilies: Schistocera vaga, S. gregaria, Gastrimargus africanus, Trimerotropis pallidipennis, and Arphia conspersa.


Separation of primary and secondary cardiovascular events in systemic anaphylaxis. The purpose of this investigation was to differentiate primary cardiac participation in systemic anaphylaxis from a cardiac reaction secondary to respiratory distress. Hemocyaninsensitized guinea pigs were anesthetized with sodium pentobarbital and artifically ventilated. The chest was opened and the left ventricle cannulated. The electrocardiogram, bronchial resistance, arterial blood pressure, and left ventricular pressure and its first derivative were recorded. Following intravenous administration of antigen, the sinus rate increased by about 50-60 beats/min, left ventricular dP/dt increased by a factor of 3, and mean arterial pressure doubled. Conduction disturbances occurred in all of the experiments and ventricular fibrillation in four of six. These changes were concomitant with a 4-fold rise in bronchial resistance. To separate the cardiac and respiratory components, antigen was administered directly into the left ventricle to expose the heart to antigen before the lungs. The intracardiac challenge resulted in increases in sinus rate and left ventricular and arterial pressure quantitatively similar to changes recorded from guinea pigs after the intravenous challenge. However, all these changes preceded the rise in bronchial resistance by 60 seconds. Arrhythmias occurred as frequently as with the intravenous challenge. Our findings show that by use of an appropriate route for administration of antigen, cardiovascular and respiratory components of systemic anaphylaxis can be separated. Our data also indicate that anaphylactic cardiovascular changes can be dissociated temporally into two sets of events: an initial primary cardiac reaction caused by intracardiac release of histamine and a subsequent cardiovascular reaction secondary to systemic release of mediator.


alpha1-Antichymotrypsin interaction with cationic proteins from granulocytes. Human granulocytes contain cationic proteins with chymotrypsin-like activity. These proteases showed a higher relative affinity for alpha1-antichymotrypsin than for alpha1-antitrypsin but the highest affinity for alpha2-macroglobulin. The complexes between cationic protein and alpha1-antichymotrypsin migrate as beta-globulin on agarose gel electrophoresis.


Intracellular localisation of 5alpha-dihydrotestosterone in human benign prostatic hypertrophy. When slices of benign hypertrophied human prostate and abdominal muscle were incubated with either [3H]testosterone or 5alpha-dihydro[3H]testosterone, the uptake of radioactivity by prostatic tissue was significantly higher than that of the muscle (P less than 0.01). The uptake of labelled androgen by prostatic tissue could be significantly reduced by adding the unlabelled steroid to the incubation medium. After the incubation of prostatic tissue with 5alpha-dihydro[3H]testosterone, the amount of the radioactivity taken up by the whole homogenate and the nuclear preparation of the prostatic tissue were measured. DNA content of the nuclei and the whole homogenate was also estimated. The mean+/-S.E.M. of 5alpha-dihydrotestosterone associated with the nuclei was 65+/-4.4%, ranging from (52.2-79.8%). The activity of acid phosphatase was measured in 30 samples of prostatic tissue. The mean +/- S.E.M. was 20.7+/-1.5 U/g tissue (9.8+/-0.9 U/mg DNA). The correlation between the activity of this enzyme and the uptake of androgen by prostatic tissue is evaluated.


Influence of hyposensitization of ATP level and CO2 production of mast cells in anaphylaxis. Anaphylaxis in a glucose-free medium containing pyruvate caused a release of histamine and a significant decrease in the ATP level of rat mast cells. The fall was maximal after 10 min and it was found to reverse after 22 min. Glucose completely counteracted the ATP fall without changing the anaphylactic histamine release. Furthermore, the oxidative metabolism of exogenous pyruvate to CO2 was stimulated in the mast cell. A high level of protection of mast cells to antigen challenge was obtained following hyposensitization and only a small amount of the intracellular histamine was released in contrast to non-hyposensitized cells. Hyposensitization counteracted the ATP fall by antigen challenge but the increase in oxidative metabolism remained unchanged. The results indicate that hyposensitization exerts effects in the mast cell consistent with a reduced ATP utilization or with a reduced uncoupling of oxidative phosphorylation. The mechanism of the hyposensitization must be due to inhibition of one or more of the cellular steps leading to histamine release and subsequent morphological changes of the cell or to uncoupling of oxidative phosphorylation.


The effects of BCG on both cellular and humoral immunity during the early response to a hapten carrier complex. Cell-mediated reactions to the carrier and antibody-mediated reactions to hapten and carrier were studied in guinea-pigs treated with a single i.v. injection of BCG at different intervals of time before immunization with two different doses of a hapten-carrier complex. The results clearly show that BCG-induced delayed hypersensitivity reactions to the carrier and decreased antibody synthesis to the hapten. It is proposed that BCG acts on a dispatcher cell which controls both cellular and humoral immunity.


Purification and characterization of human liver specific F antigen. This paper reports the properties of purified human F antigen (liver-specific antigen). Homogenates of liver in 0-25 M sucrose were centrifuged at 105,000 g. The supernatants were chromatographed on Sepharose 6-B and four major peaks were separated. The third peak proved to be predominantly F antigen. This fraction was subsequently subjected to DEAE-cellulose column chromatography and F antigen was eluted at a concentration less than 0-2 M NaCl in 0-01 M sodium phosphate buffer (pH 7-2). Finally, purified F antigen was obtained after preparative isoelectric focusing. Purified human F antigen was found to have a mol. wt between 40,000 and 80,000, a pI of 6-5-6-7 and a density of 1-26. It is a protein antigen and contains no detectable carbohydrate or lipid. No differences were found in purified F-antigen preparations from several species when tested by sodium dodecyl sulphate (SDS) disc gel electrophoresis. Immunofluorescent studies showed that F antigen was homogeneously distributed in the cytoplasm of liver cells but was not present on the cell surface. Immunization of guinea-pigs with purified human liver-specific protein did not induce antibody to the F antigenic determinant defined by mouse anti-F antiserum. It did, however, induce antibodies to two human liver antigens. One of these seems to be a human-specific determinant on the F antigen molecule and the other appears to be a separate molecule which is similar in molecular weight and electrophoretic mobility to the F-antigen molecule.


Studies on the specificity of smooth-muscle antibodies. Purified contractile proteins from smooth and striated muscles have been used to test the specificity of human smooth muscle antibodies (SMA) from patients with chronic liver disease (IgG-SMA) and acute hepatitis (IgM-SMA). The reactions, as detected by indirect immunofluorescence, of IgG-SMA with renal vessel walls, renal glomeruli, peritubular fibrils and the luminal part of the tubular cells could be completely abolished by absorption with either smooth muscle or skeletal muscle F-actin, while absorption with myosin and tropomyosin had no effect. The specificity of IgG-SMA for actin was confirmed by their staining of the actin-rich I-bands of skeletal muscle myofibrils and by the blocking of this reaction by pretreatment of myofibrils and isolated smooth muscle cells with smooth muscle myosin subfragment 1 (S-1). IgM-SMA from patients with acute hepatitis-stained renal vessel walls and some sera also stained renal glomeruli. The IgM-SMA titres decreased after absorption both with myosin and F-actin but not with tropomyosin. The reactivity of some IgM-SMA could be blocked by S-1 while others could not. Thus the specificity of IgM-SMA seemed to be variable, and apparently differed from IgG-SMA in some cases.


Cytophilic antibodies in bronchopulmonary aspergilloma and cryptogenic pulmonary eosinophilia. The immunoglobulin class and subclass of cytophilic antibodies have been studied using peripheral leucocytes from twenty-two patients with allergic bronchopulmonary aspergillosis, aspergilloma and cryptogenic pulmonary eosinophilia. In patients with allergic bronchopulmonary aspergillosis, significantly increased histamine liberation occurred following challenge of their leucocytes with antisera to IgE, IgG2, IgG3 and IgG4 as well as with Aspergillus fumigatus antigen. The results were considerably modified if the patient was receiving corticosteroids at the time of the test. The presence of IgG2-specific antibody to A. fumigatus in the serum of one patient, capable of sensitizing donor leucocytes, was demonstrated in passive sensitization experiments. In two patients with uncomplicated aspergillomas no evidence of cytophilic antibody to any class was found although large amounts of precipitating IgG antibody was present in the serum. Two patients with aspergilloma and systemic symptoms of weight loss and fatigue (which have been interpreted by others as 'hypersensitivity' responses) had increased amounts of cytophilic antibody similar to those with allergic bronchopulmonary aspergillosis. Six patients with cryptogenic pulmonary eosinophilia were also studied. No evidence of specific antibody to A. fumigatus was found but, as a group, significantly increased histamine liberation using antisera to IgG2 was demonstrated. Individual patients also showed evidence of other classes of cytophilic antibody, one having IgE, three IgG3 and two IgG4. The relationship between heat-stable short-term sensitizing antibody (IgG STS) inducing immediate skin responses and the pattern of cytophilic antibodies found in our patients with bronchopulmonary aspergillosis having dual (immediate and late reactions) is discussed. Clinically these tests are of diagnostic value and they may be helpful in assessing symptomatic patients with aspergillomas for corticosteroid treatment.


The Ia antigens.

The Ia antigens. The Ia antigens constitute a polymorphic series of cell surface determinants. At present, their definition is mainly a genetic one, and thus any cell surface antigen which can be demonstrated to be encoded by a gene in the Ir region of the H-2 complex may be classified as an Ia antigen. There are presently three subregions of the I region defined on the basis of available recombinant haplotypes, and designated at I-A, I-B, AND I-C. Mapping of individual Ia specificities indicates that numerous specificities are determined by genes in the I-A subregion, several in the I-C subregion, and few, if any, in the I-B subregion. This may be a reflection of the state of the art, however, rather than an accurate assessment of the extent of polymorphism. The Ia antigens appear to be expressed preferentially on the B-cell subpopulation of lymphoid cells. However, with the use of sensitive techniques they have also been demonstrated on some T cells, on macrophages, on sperm cells, and on epidermal cells. The Ia antigens have also been demonstrated on several T-cell factors which appear to be involved in the immune response. Whether or not all of the Ia antigens thus localized are identical or represent overlapping specificities within the same sera remains in many cases to be determined. There are presently three ways of defining Ia specificities serologically: (1) by direct immunization between strains differing only in the I region; (2) by detection of shared Ia determinants using polyspecific sera which contain H-2K region and H-2D region antibodies but which are nevertheless specific only for Ia antigens when tested on target cells of other strains; and (3) by selective absorption of H-2K region and H-2D region antibodies from an H-2 antiserum by cells bearing these antigens but lacking (or relatively lacking Ia antigens. All three of these methods produce anti-Ia reagents of reasonable titer for use in both serological and functional experimentation. The definition of the specificity as an Ia specificity in each case requires the availability of appropriate recombinant strains to map the specificity to the Iregion. In addition, there are several correlative criteria which have been developed in order to detect Ia activity in alloantisera in the absence of the availability of appropriate recombinants for mapping of the specificity. These include the tissue distribution of the Ia antigens (namely, their predominant expression on the B-cell subpopulation), their characteristics molecular size, their association on the B-cell surface with the Fc receptor, and their  lack of association with other products of the major histocompatibility complex as distinguished either chemically or by cocapping studies. These correlative criteria make it possible to distinguish probable anti-Ia reactivity in a variety of serological reactions, but the results must still be interpreted with caution until appropriate recombinants have been obtained which can map the specificities to the I region...


Preliminary amino acid sequences of transplantation antigens: genetic and evolutionary implications. Preliminary amino acid sequence data on the transplantation antigens of mouse and man have led to provocative hypotheses about the genetic organization and evolution of genes coded by the major histocompatibility complex of mammals. New microsequencing techniques should permit a detailed analysis of these gene products and an eventual choice among the alternative hypotheses now posed. These data have made it apparent that the H-2 complex is  a fascinating and complicated chromosomal region which will continue for some time to intrigue immunologists, geneticists, biochemists, and cell biologists.


Serum protein synthesis in mutant mice with abnormal hepatic endoplasmic reticulum. Severe ultrastructural abnormalities of liver endoplasmic reticulum have been described in newborn mice homozygous for radiation-induced deletion alleles at the colour locus. The ultrastructural defects were accompanied by deficiencies of several enzymes and lowered serum protein levels. Studies on serum protein synthesis were undertaken to see if decreased rates of synthesis, especially of constituents thought to be synthesized on membrane-bound ribosomes, were the cause of the deficiencies. Although decreases or absence of several serum proteins were shown, radiopulse-immunoprecipitation studies of albumin and fibrinogen synthesis suggested that the decreased synthesis rates were a secondary defect. Serum glycoproteins were not altered more than other constituents in the mutant material.


Negotiating educational programs for children with developmental disorders: assessment, interpretation, demonstration, support (AIDS). An approach to negotiating educational programs for children with multiple developmental problems is presented using two case examples from an interdisciplianry setting, the University Affiliated Cincinnati Center for Developmental Disorders. The use of the AIDS (Assessment, Intervention, Demonstration, Support) approach offers the schools evidence that the professional educator and psychologist are working in good faith and will not abandon them prematurely. Proper school placement through mediation also allows the school personnel to receive credit from the family for making the necessary changes, therefore not losing face with the parents. This approach seems particularly applicable to children with multiple and chronic problems who do not fit single special educational/diagnostic categories or labels.


Detection of sister chromatid exchanges by 4'-6-diamidino-2-phenylindole fluorescence. This paper describes a 4'-6-diamidino-2-phenylindole (DAPI) fluorescent technique for differentiation of sister chromatids and for the study of sister chromatid exchanges (SCE) in mouse chromosomes. The advantages of the DAPI fluorescent technique are also described. Differences in the occurrence of SCE between the centromeric heterochromatin (C-banded) and the chromosomal arm chromatin were studied in mouse cells (RAG) with or without mitomycin C treatment. Single strand exchanges between the DNA double helices in the sister chromatids were not detected. SCE and chromosome breakage appeared to occur more frequently in the centromeric region than in the chromosomal arm. This might play an important role in chromosome evolution in mice.


Characterization of Drosophila heterochromatin. I. Staining and decondensation with Hoechst 33258 and quinacrine. A number of preliminary experiments have shown that the fluorescence pattern of Hoechst 33258, as opposed to that of quinacrine, varies with the concentration of dye. The metaphase chromosomes of D. melanogaster, D. simulans, D. virilis, D. texana, D. hydei and D. ezoana have therefore been stained with two concentrations of H 33258 (0.05 and 0.5 mug/ml in phosphate buffer at pH 7) and with a single concentration of quinacrine (0.5% in absolute alcohol). The three fluorescence patterns so obtained were shown to be somewhat different in some of the species and the coincide in others. All three stainings gave an excellent longitudinal differentiation of heterochromatin while euchromatin fluoresced homogeneously. Living ganglion cells of the six species mentioned above were treated with quinacrine and H 33258. Quinacrine induced a generalized lengthening and swelling of the chromosomes and H 33258 the decondensation of specific heterochromatic regions. A correlation of the base composition of the satellite DNAs contained in the heterochromatin of the species studied with the relative fluorescence and decondensation patterns showed that: 1) the extremely fluorochrome bright areas and those decondensed are present only in species containing AT rich satellite DNA; 2) the opposite is not true since some AT-rich satellite DNAs are neither fluorochrome bright nor decondensed; 3) there is no good correspondence between Hoechst bright areas and the decondensed ones. AT richness therefore appears to be a necessary but not sufficient condition both for bright fluorescence and decondensation. Some cytological evidence suggests that similarly AT rich satellite DNAs respond differently in fluorescence and decondensation because they are bound to different chromosomal proteins. A combination of the results of fluorescence and decondensation revealed at least 14 types of heterochromatin; 4-7 of which are simultaneously present in the same species. Since closely related species (i.e. D. melanogaster and D. simulans; D. virilis and D. texana) show marked differences in the heterochromatic types they contain, it can be suggested that within the genus Drosophila qualitative variations of heterochromatin have played an important role in speciation.


Characterization of Drosophila heterochromatin. II. C- and N-banding. The C- and N-banding patterns of D. melanogaster, D. simulans, D. virilis, D. texana, D. ezoana and D. hydei were studied in comparison with quinacrine and Hoechst banding patterns. In all these Drosophila species the C bands correspond to the heterochromatin as revealed by the positive heteropycnosis in the prometaphase chromosomes. The N bands have the following characteristics: 1) they are always localized on the heterochromatin and generally do not correspond to the C bands; 2) they do not correspond to the nucleolar organizing regions; 3) they are inversely correlated with fluorescence, i.e., they correspond to regions which are scarcely, if at all, fluorescent after Hoechst 33258 or quinacrine staining; 4) they are localized both on regions containing AT rich satellite DNA and on those containing GC rich satellite DNA.


Sequence of DNA replication in 277 R- and Q-bands of human chromosomes using a BrdU treatment. Replication times for all important chromosome bands, of both types R and Q (277 structures) are analysed. The R-bands form a group of structures whose DNA replicates during the early S-phase, while the DNA situated in the Q-bands replicates during the late S-phase. There may not exist overlapping between replication times of these two types of structures. The widest R-bands are those which are the earliest to replicate; in general, the most intense Q-bands are those which are the latest to replicate. Especially among these last ones, a certain asynchronism exists between the replication times. Finally the heterochromatin of chromosomes 1, 16 and Y and of the short arms of the acrocentrics could contain two types of DNA which replicate at different times.


Characterization of the toromeric structure in Drosophila lummei. The toromere, a structure previously reported only in several strains of Drosophila melanogaster, is found in salivary gland nuclei of three populations of Drosophila lummei, a member of the virilis group. The toromere is characterized by being quinacrine-bright and Feulgen-positive. Further staining and enzyme digestion procedures reveal that the toromere is composed of double-stranded DNA with little or no protein complexed with it. The toromere appears as a small quinacrine-bright dot in diploid cells and apparently undergoes polytenization, as it is observed as a large quinacrine-bright doughnut-shaped structure in salivary gland preparations. We describe the effect of larval culture temperature on the morphology and number of toromeres per cell, and lend support to the proposal that the toromere could be a highly replicated episome.


Dotted chromosomes produced in sodium phosphate solution supersaturated with NaHCO3. Dotted chromosomes were consistently produced in both BrdU and non-BrdU substituted Chinese hamster cells after treatment with 1.0 M Na-phosphate solution, adjusted to pH 9.0 with a supersaturating amount of NaHCO3, and at a temperature of 80--95 degrees C. -- A series of changes in chromosome morphology was produced as the temperature of the solution was progressively increased. In BrdU-treated cells, G-banding and differentially stained sister chromatids were sequentially produced prior to the appearance of dots. In non-BrdU treated cells, only G-banding was produced before dot formation. In general, the patterns of dots correspond to the G-banding patterns. --Chromatids, with uni- or bifilarly BrdU substituted DNA or with normal DNA, required differential temperatures for the production of dots. Since the temperature required for dot formation was always slightly higher than that required for producing differentially stained chromatids, this phenomenon can be used as an important indicator for determining the optimal temperature required for revealing differentially stained chromatids.


In vivo BrdU-33258 Hoechst analysis of DNA replication kinetics and sister chromatid exchange formation in mouse somatic and meiotic cells. BrdU (5-bromodeoxyuridine)-33258 Hoechst methods have been adapted for in vivo analyses of replication kinetics, sister chromatid differentiation and sister chromatid exchange (SCE) formation in mice. Sufficient in vivo BrdU substitution for cytological detection was effected with multiple intraperitoneal injections of the analogue. The combination of centromere staining asymmetry and sister chromatid differentiation at metaphase permits unambiguous determination of the number of replications in BrdU and dT (deoxythymidine) undergone by individual cells. Late-replicating regions in marrow and spermatogonial chromosomes are highlighted by bright fluorescence after sequential incorporation of BrdU followed by dT during a single DNA synthesis period. SCEs are analyzed in marrow and spermatogonial metaphases after successive complete cycles of BrdU and dT incorporation. Significant induction of SCE was observed with both mitomycin C and cyclophosphamide; the latter drug requires host-mediated activation to be effective. In meiotic metaphase cells harvested two weeks after BrdU incorporation, satellite DNA asymmetry, sister chromatid differentiation and SCE could be detected in a few chromosomes, most frequently the X and the Y.


Xerophthalmia and measles in Kenya.

Xerophthalmia and measles in Kenya. In many African countries measles is considered to be an important cause of blindness. On the basis of his observations in Kenya and Tanzania in 1972 Franken presumed, however, that in the majority of these cases xerophthalmia was the real cause of blindness, precipitated by the "catalyst" measles. In order to gain a better understanding of this important complicated problem, we performed in the first half of 1974 an investigation in Kenya into the prevalence of xerophthalmia. In December 1974 we had the opportunity to evaluate our Kenyan findings on Java, in the company of Dr. J. ten Doesschate and Professor H.A.P.C. Oomen. The results of this investigation in Kenya and Indonesia are presented in this thesis. (see article) 1. Xerophthalmia occurred nearly everywhere in Kenya in 1974. This demonstrates the prevalence of xerophthalmia in communities which - do not have rice but - have maize for their staplefood. 2. Xerophthalmia appears to be the main cause of blindness in Kenyan children. 3. Measles often plays - by means of local and general "catalysing" effects - an important role in the development of blindness caused by xerophthalmia. 4. In well-nourished children measles is of no consequence as a cause of blindness. 5. Vital staining by 1% rose bengal or 1% lissamine green appears to be a real asset for the early diagnosis of xerophthalmia in Health Centres and in field surveys. This method is therefore of great importance for the prevention of severe, blindness inducing vitamin A deficiency.


[Principles of programmed teaching].

[Principles of programmed teaching]. Student education and continued education programs increase the stress on a dental school teacher as far as time is concerned. The use of modern aids gives an effective relief, if existing teaching materials in basic fields can be taken over. Quality characteristics and basics of the evalutation of teaching programs are presented.


Nature of cross-reaction between hCG and anti-oLH serum and development of a radioimmunoassay to measure hLH specifically in the presence of hCG. Immunological cross-reaction between hCG and anti-oLH sera has been demonstrated using radioimmunoassay techniques. The results indicate that this cross-reaction is incomplete and that the anti-oLH sera used have the ability to distinguish between LH and hCG. Following absorption with purified hCG, anti-oLH serum was used to develop a heterologous radioimmunoassay "[125I]iodo-hLH + anti-oLH serum" (H-O, RIA) which specifically and selectively measures hLH in serum samples containing both hLH and hCG. In this radioimmunoassay hCG and subunits of hCG do not cross-react with hLH, in the range in which these hormones are present in human serum under physiological conditions. Other hormones such as hPL, hPRL, hGH, hFSH, hTSH, and GnRH do not interfere with the measurement of LH by radioimmunoassay. The sensitivity of the assay was 1.5 mIU (25 ng) per ml (LER 907 standard), and the inter- and intra-assay coefficients of variations for samples were 10.83% and 8.4%, respectively. The recoveries of hLH added to pregnancy serum containing an hCG concentration of 8.55 IU/ml were in the range 95-108%. Determination of LH content of human pituitary extracts by H-O RIA gave values which were in close agreement with those derived by bioassay (indices of discrimination 0.72-1.12). Serum LH patterns in women during normal menstrual cycles as well as in amenorrheic patients who received GnRH treatment are comparable to those reported by other investigators using other radioimmunoassay systems. Serum samples obtained during the first trimester of pregnancy, when analyzed by H-O RIA, showed basal LH levels.


Quinidine syncope: torsade de pointes with low quinidine plasma concentrations. In 2 patients without clinically significant ischemic heart diseases, oral quinidine was used to control supraventricular arrhythmias. In both patients, syncopal attacks occurred, caused by a particular type of ventricular tachyarrhythmia called torsades de pointes. Quinidine plasma concentrations were low (2.6 and 1.2 mg/1, respectively); QRS duration was normal, but the Q-T interval was markedly prolonged.


Increased plasma binding of quinidine after surgery: a preliminary report. The plasma binding of quinidine and phenytoin has been studied pre- and postoperatively in nine patients submitted to planned gastric surgery. The binding of phenytoin showed a slight and transient reduction, whilst quinidine binding was markedly increased, on average from 78.5% on the day of operation to a maximum of 87.5%, after 2-4 days. The time course of the increase was strikingly parallel to that of the concentration of certain acute phase proteins.


Blockade of specific antibody-forming cells in vivo by dextrans and levans. Fractions of dextrian (DE) B512, DE B1355 and levan (LE) have been shown to induce a specific blockade of antibody-forming cells (AFC) when injected into previously immunized mice. Whereas specific blockade with DE B512 (450 000 daltons) was easily induced by as little as 10 mug, blockade to LE and DE B1355 was more resistant and required 1 and 10 mg, respectively. AFC blockade and tolerance are dissociable phenomena, as the former effect could be achieved with nontolerogenic low mol. wt DE B512 (10 000 daltons). Conversely, perennial rye grass fructosan (7600 daltons), which is tolerogenic for LE, did not induce a blockade. Furthermore, blockade of anti-DE response specific for alpha (1 leads to 3)-linked glucosyl determinants was achieved in BALB/c mice, whereas attempts to induce stable tolerance have been unsuccessful.


HLA-B specificities and w4, w6 specificities are on the same polypeptide. The human alloantigenic specificities w4 and w6, which are products of a diallelic system genetically associated with the HLA-B locus, have been solubilized by papain digestion of membranes from the lymphoblastoid cell line, RPMI 4265. The w4 and w6 specificities copurified with the HLA-B locus products, HLA-B7 and HLA-B12. Sequential immunoprecipitation experiments were performed using alloantisera to HLA-B7, HLA-B12, w4 and w6, and a purified HLA-B7, B12, w4, w6 antigen pool labeled with 125I-Bolton-Hunter reagent. These experiments demonstrated directly that HLA-B7 and w6, which are genetically associated with each other, are different antigenic determinants on the same molecule, while HLA-B12 and w4, also genetically associated, are distinct antigenic determinants on a second molecule. Arguments are presented which suggest that the HLA-B determinants and w4, w6 determinants are in fact on the same polypeptide, and the genetic implications of the findings are discussed.


Antigen, host and adjuvant requirements for induction of hyperacute experimental autoimmune encephalomyelitis. A hyperacute form of experimental autoimmune encephalomyelitis (HEAE) was induced in Lewis rats using small doses (3.2 mug) of guinea pig myelin basic protein as immunogen and B. pertussis vaccine as adjuvant. Myelin basic proteins from species other than guinea pig (rat, man, monkey, pig, ox, rabbit and sheep) induced only ordinary EAE with this adjuvant. HEAE was more readily distinguished from ordinary EAE by clinical criteria (early onset, with a rapid and severe course, and high incidence of cerebral signs and mortality) than by histologic signs which, although characteristic of HEAE. were not pathognomonic for HEAE, HEAE was transferred to x-irradiated syngeneic recipient rats with lymph node cells from appropriately immunized donors. The Brown Norway (BN) strain of rat was found susceptible to induction of ordinary EAE, but not HEAE, using large doses of either rat or guinea pig myelin basic proteins. The unique immunogenicity of the guinea pig basic protein must be due to a different antigenic determinant from the determinant(s) which is shared by rat and guinea pig myelin basic proteins and which without B. pertussis induces ordinary EAE. The adjuvant action of B. pertussis in inducing HEAE in the Lewis rat is most likely mediated through an immunocompetent T lymphocyte.


Effect of prior sensitization with hapten on the antibovine IgG antibody response to hapten-conjugated tolerogen of mice tolerized by low doses of bovine IgG. Mice tolerized by low doses of bovine IgG (BGG), immunized with arsanilated (ARS) conjugates and challenged with ARS-BGG show an augmented anti-BGG antibody response compared with controls immunized with nonconjugated carrier. Conditions optimal for generating hapten help are explored and suggest that the phenomenon is mediated by hapten-specific helper T cells. During exploration of these conditions it became apparent that the response was suppression rather than augmentation with ARS, under certain circumstances, and with 2,4-dinitrophenyl, 4-hydroxy-5-iodo-3-nitro-phenylacetyl (NIP) and sulfanilic acid, under all conditions tested. Especially important in determining the outcome of the interaction was the type of adjuvant used for hapten priming. The significance of these observations is discussed in relationship to the balance between negative and positive cooperation.


Beta2-microglobulin distribution in human normal tissues. The distribution of beta2-microblobulin in human normal tissues was investigated by the indirect immunofluorescent antibody method. Lymphoid, macrophage and endothelial cells were consistently positive in every organ studied. In addition, only the stratum germinativum of the epidermis, some tracts of the columnar epithelium of the digestive system and some endometrial tubular glands showed a specific fluorescence.


Naturally-occurring double-stranded RNA and immune responses. IV. Influence of molecular size on antigenicity and adjuvant activity. The immunological properties of a naturally-occurring double-stranded ribonucleic acid (ds-RNA), obtained from a mycophage of Penicillium chrysogenum, have been studied in relation to molecular size. Materials of reduced size, as reflected by molecular weight measurements, produced by ultrasonication of native ds-RNA, exhibited progressively lowered ability to induce an anti-ds-RNA response in mice. Adjuvant and immunosuppressive activities were of similar magnitude in both high and low molecular weight fractions. Evidence was also obtained of increased toxicity in materials of reduced size.


Antigenic determinants of heavy chain variable regions: immumological typing of the human immunoglobulin VHIII subgroup. An antigenic determinant of the VHIII variable region subgroup was defined by means of a heterologous specific antiserum using a hemagglutination inhibition procedure. The specificity of this antiserum was established in inhibition experiments with proteins either of known primary structure or belonging to a definite VH subgroup. A series of IgG, IgA, IgM and IgD monoclonal proteins was examined for the presence of this VHIII subgroup antigenic determinant. The data showed that 50% of the IgG, 62% of the IgA, 55% of the IgM and 41% of the IgD were VHIII-positive, and that certain "blocked" monoclonal immunoglobulins belonged to this subgroup. A preferential association of the VHIII antigenic determinant with the IgG1 and IgG3 subclasses was observed among IgG myeloma proteins while the preferential association was only observed with the IgG1 subclass when anti-Rh antibodies were studied. The VHIII subgroup exhibited nonallelic behavior.


Shared determinants between virus-infected and trinitrophenyl-conjugated H-2-identical target cells detected in cell-mediated lympholysis. Infection of H-2-identical mice with either lymphocytic choriomeningitis (LCM) virus, vaccinia virus, or paramyxo (Sendai) virus resulted in the generation of specifically sensitized cytotoxic T lymphocytes (CTL). CTL generated in vitro against 2,4,6-trinitrophenyl (TNP)-conjugated syngeneic stimulator cells were specifically cytotoxic for TNP-conjugated H-2K (D) region identical targets. Both LCM and vaccinia-induced CTL, however, were found to be strongly cytotoxic towards TNP-conjugated, H-2K(D) region-identical target cells. In contrast, Sendai virus-induced CTL did not lyse TNP-conjugated, syngeneic target cells. Inhibition experiments using cold targets suggested that shared antigenic determinants can be detected on either LCM virus-infected and TNP-conjugated targets, which are not present on the cell surface of normal target cells.


Role of epitope density in the induction of tolerance and immunity with thymus-independent antigens. III. Interaction of epitope density and receptor avidity. The induction of B cell tolerance to 2,4-dinitrophenyl conjugates of polysaccharide antigens (levan or dextran) was studied in mice primed with keyhole limpet hemocyanin (KLH) or 2,4,6-trinitrophenylated KLH. The relationship of the epitope density of the tolerogen with avidity of B cell receptors (as judged indirectly by a plaque inhibition assay) was investigated. It was found that high avidity precursors (IgG) were tolerized by antigen of much lower epitope density, and at lower concentration, than were low avidity precursors (especially IgM cells). IgA cells were intermediate in behavior. These results suggest that the epitope density effect acts by ensuring a necessary degree and/or energy of antigen binding.


Evidence for an absence of cross-reactivity between the human beta2-microblobulin and human IgG allotypes and subclasses. Six beta2-microglobulins were isolated from the urine of six patients with renal tubular dysfunction. Cross-reactivity between these beta2-microglobulins and antigenic determinants of different IgG allotypes and subclasses was investigated. No evidence of cross-reactivity was found. Some hypotheses are discussed concerning these results.


Synthesis and release of histamine studied on slices from rat hypothalamus. In slices from rat hypothalamus, incubated in the presence of 3H-L-histidine (3H-L-His), the aminoacid was rapidly taken up by a saturable process, and partially converted into 3H-histamine (3H-HA). The overall conversion was prevented either by inhibitors of L-histidine decarboxylase or by aromatic aminoacids competing with L-His uptake. The synthesis process exhibited Michaelis--Menten kinetics with an affinity of the aminoacid not different from that for the decarboxylase in homogenates; however the Vmax in homogenates was more than 10 times higher than in slices. Depolarization of the slices by 50 mM potassium resulted in: (a) a calcium-dependent release of 3H-HA which was more marked than that previously reported for endogenous HA, (b) a significant acceleration in the rate of 3H-HA synthesis, which was characterized by an unchanged Km but a significantly elevated Vmax. The regulation of HA synthesis did not appear to depend on end-product inhibition since it was not midified by the addition of exogenous HA. The release of 3H-HA was followed by the accumulation of 3H-methylhistamine, which was enhanced by a monoamine oxidase inhibitor. Aminoguanidine, a diamine oxidase inhibitor, had no effect on catabolism. The involvement of mast-cells in the storage of a fraction of endogenous HA in hypothalamic slices was assessed by the significant releasing effect of compound 48/80. Hence, the data support the existence of two distinct HA stores in the brain: depolarization relases the amine and increases its synthesis, probably in neurones, whereas compound 48/80 releases it from a slowly turning-over store, probably in mast-cells.


The retinal projection to the ventral part of the lateral geniculate nucleus. An experimental study with silver-impregnation methods and axoplasmic protein tracing. The retinal projection to the ventral part of the lateral geniculate nucleus (LGNv) was studied in 25 adult cats. In 12 cats one or both eyes were enucleated and the terminal degeneration in the LGNv was studied with silver impregnation methods. In 12 cats one or both eyes were injected with 3H-leucine and the terminal labelling in the LGNv was studied with autoradiography. In one animal one eye was enucleated and the other injected. In this case alternate sections were silver impregnated and processed for autoradiography. The series were cut in parasagittal, transverse or horizontal planes. The results revealed by degeneration were in very good agreement with those revealed by axoplasmic protein tracing. Two fields of retinal projection were found in the LGNn. The larger, dorsal one was restricted to subnucleus k (Jordan and Holländer, 1972) and comprised an ipsilateral and a contralateral component which did not overlap each other. The smaller, ventral field of projection was contralateral and extended from subnucleus b into c, e and a. The afferent optic tract fibres to both terminal fields were of small calibers.


The climbing fibers of the cerebellar cortex, their origin and pathways in cat. The sources and pathways of the climbing fibers to the cerebellar posterior vermis were studied with comibined electrophysiological and anatomical methods in cats. Recording from identified cerebellar Purkinje cells, monosynaptic climbing fiber (CF) responses have been obtained both for stimulation of the inferior olive (IO) and various parts of the brain stem (BS). CF responses were found to of three types, IO only, BS only or both IO and BS. However the responses to BS stimulation were very few in number in comparison with IO or IO and BS types of responses. The latencies of the responses were shorter for the BS cases consistent with their distance from the cerebellum. A comparison of latencies and the relative responsiveness of the different area of the brain stem which were studied, indicate that part of the CF ascend through the pontine region and enter the cerebellum by way of the medium and superior penduncles. This finding is confirmed by the results of anatomical studies in which degenerating fibers were found in the molecular layer (using the Nauta technique) after lesion of the brachium pontis but not after lesions of the medial portion of the pons. Similarly, injection of radioactive leucine into the pontine nuclei failed to show any labeled fibers in the molecular layer. Horseradish peroxidase (HRP) was injected into localized regions of the posterior vermis after total bilateral destruction of the inferior peduncles. Large numbers of positive, marked cells were still found in the inferior olive. It is concluded that nearly all, if not all, the climbing fibers originate in the inferior olive and that they ascend to the cerebellum by way of all the peduncles.


Immunological heterogeneity in human ferritinemia. Hyperferritinemia in various diseases, mainly hematological, was confirmed by immunological methods. For ferritin detection, anti-human placental ferritin antiserum, anti-human hepatic ferritin antiserum, and anti-human leukemia cell ferritin antiserum were used and the result was compared with each other. Leukemia, malignant lymphoma, multiple myeloma, and aplastic anemia are hematological diseases which showed a positive reaction in this test, among which leukemia showed the highest positivity. Cases of hepatic diseases and non-hematological malignant neoplasms also showed a positive reaction. The positivity was quite low and almost negligible in other diseases and healthy individuals. Anti-human placental ferritin antiserum seemed to be suitable for cancer diagnosis and, antihuman hepatic ferritin antiserum for hepatic diseases. The results of analysis of purified human hepatic and placental ferritins highly suggested the presence of immunological heterogeneities between them. Also, a possibility was pointed out that one of the components of the so-called leukemia-specific antigens might sometimes be the isoferritin of leukemia cells.


Determination of the bleomycin-inactivating enzyme in biopsies. A method for the determination of the Bleomycin-inactivating enzyme is described. The amount of conversion of active Bleomycin to inactive Bleomycin is used as a measure for the determination of the Bleomycin-inactivating enzyme activity. The active Bleomycin is determined in a DNA-dependent DNA polymerase assay; in a certain Bleomycin concentration range the resulting reduction of the activity of the DNA-dependent DNA polymerase is correlated linearly when plotted semilogarithmically. By application of this method it has been found that the activity of the Bleomycin-inactivating enzyme is highest in liver and present in lower concentrations in testis, spleen, lung, brain, and skin.


[Successes and prospects for genetic engineering]. The review of literature (1970-1976) on problems of gene engineering is given. Gene engineering is pointed out to be a new method of modern biology and a new page of modern molecular genetics. Gene engineering detected a real possibility of artificial creating living hybrid organisms, i.e. constructing functional recombinant DNA molecules according to a project of investigator, but not to possibilities of crossing. The determination of gene engineering (in contrast with genetical engineering) is given in the first division of the article. Genetical engineering is a construction of hybrid organisms on the basis of recombination between non-homologous chromosomes cy crossing. Genetical engineering is based on sex crossing, thus the application of this method is restricted by crossability (i.e. experiments in vivo), which possibilities are determined by taxonomical limits. Gene engineering is a new method of operating directly with genes. It permits constructing in vitro any hybrid genomes desirable. There is no limits of combining ability for gene engineering. Three main stages of constructing hybrid genomes should be taken into account for the proper determination of gene engineering as a method of genome constructing: 1) the gene isolation; 2) their cross-linking in vitro; 3) the transfer of hybrid DNA into recipient cell or its genome. The cardinal stage of gene engineering is the construction of hybrid DNA, cross-linking any initial DNAs from any remote animals, plants and bacteria. All the methods known of gene isolation are described. The chemical method of gene isolation is based on that case, when DNA of some gene differs in its physico-chemical characteristics from total DNA, for example, DNAs of genes coding ribosomal RNAs or sea urchine histone DNA. Isolation of promotors and operators using DNA dependent RNA polymerase, which recognizes promotors, repressor and operator DNA, should also be considered as the chemical method of gene isolation. Restrictase method, which is also well known, is convenuent because the restricts have long enough sticky ends, which is important for the following gene cross-linking. The method of total restriction, reported by Lederberg et al. and Debabov et al., is described. The phage method (in particular, Shimada method) is given, permitting the direct integration of lambda phage into a number of sites of Escherichia coli chromosome. Gene engineering method of gene isolation is mentioned, in particular, the data of Kameron et al. on hybrid phages carrying DNA ligase gene, and Clark a. Carbon on hybrid plasmids carrying triptophane and arabinose operons genes. These methods are called "shot gun". Methods of gene isolation from higher organisms are less developed. A method of gene isolation using so called colony hybridization (according to Grünstein and Hognes) is also given...


Demonstration of an intracellular copper-binding protein by orcein staining in long-standing cholestatic liver diseases. Liver biopsies from eight patients with primary biliary cirrhosis, two with chronic active hepatitis of a cholestatic form, three with long-standing alcoholic liver cirrhosis, and one with extrahepatic biliary obstruction were studied. In each case dark brown cytoplasmic material was seen after staining of the tissue sections with Shikata's orcein method. In exactly the same cellular and subcellular locations as the orcein-positive material, and with morphologically equal granules, two different ordinary staining methods for copper (rubeanic acid and Mallory-Parker's haematoxylin) gave positive reactions. The earlier histochemical findings have revealed the protein nature and high sulphydryl content of the orcein-positive material. Its close association with copper in liver sections suggests its copper-binding nature and indicates that a common copper-protein complex accumulates in the cytoplasm of liver cells during longstanding cholestasis in biliary diseases of various pathogenetic origin.


Morphological types of hepatocellular hyalin in Indian childhood cirrhosis: an ultrastructural study. Morphological variants of intrahepatocytic hyalin in Indian childhood cirrhosis have been analysed by electron microscopy. This structure, morphologically identical with Mallory's alcoholic hyalin, is encountered in three different forms. The most common one is composed of randomly oriented fibrils. The next common type is composed of smudgy homogeneous or finely granular material, while the least common type consists of parallel fibrils with gentle curves giving a 'finger-print' appearance. Fragmented endoplasmic reticulum and ribosomes were frequently associated with hyalin suggesting that these organelles contribute to their formation.


Cell-mediated immunity in pregnant women. Phytohemagglutinin-induced lymphocyte transformation (PHA-T) was depressed in pregnant women, as compared with that in nonpregnant women. Pregnancy serum had a suppressive action on PHA-T which was enhanced with the advance in pregnancy. Hydrocortisone, progesterone, alpha-fetoprotein and trophoblast-specific antigen, were demonstrated as immune suppressive factors. From these results, it was concluded that cell-mediated immunity might be reduced in pregnant women and that this reduction might be one of the causes for the maintenance of pregnancy.


Correlations between radioiodine uptake and blood viscosity factors in hypothyroid and hyperthyroid states. A study of blood viscosity factors was undertaken in six hypothyroid and eleven thyrotoxic patients (females) whose thyroid function was defined by means of radioiodine uptake and protein bound (PB) 131I tests. Results indicate that: (i) significant differences exist between hypothyroid and hyperthyroid groups in correlations between radioiodine tests and blood viscosity factors (and fibrinogen levels); (ii) blood viscosity in thyrotoxic patients decreases as the degree of thyrotoxicity increases, while an opposite phenomenon is observed in hypothyroid patients, and (iii) degree of rigidity of the red cells is associated with the radioiodine uptake and PB 131I tests, the correlations being significant and negative.


An improved technique for selective silver staining of nucleolar organizer regions in human chromosomes. A reliable technique for staining human chromosomal nucleolar organizers (NOR's) with silver solutions is described. The NOR's can be selectively stained dark brown by silver solutions leaving the chromosome arms unstained and available for counterstaining with orcein or Giemsa dyes. Unequivocal identification of chromosome pairs bearing NOR's can be achieved using fluorescent banding techniques followed by silver staining. The silver staining procedure for NOR's was simplified and standardized through control of the chemical and physical conditions during silver impregnation and developing.


Production of C and T bands in human mitotic chromosomes after heat treatment. A modified thermal denaturation of human chromosomes in Hanks' solution at 91 degrees C produces highly contrasted C and T bands. This improved double-banding system is a good tool in the analysis of ring chromosomes, for which R banding is often inadequate, as it is exemplified.


Segmentation of human chromosomes induced by 5-ACR (5-azacytidine). The 5-ACR (5-azacytidine) introduced in human lymphocyte cultures induces a lack or a delay of condensation of some chromosome segments corresponding to the G-bands. The resulting R-banding is very similar to that obtained with a 7-h treatment by BrdU, although the segmentation may be much stronger (pulverization) with high doses. However, the 5-ACR does not induce chromatid asymmetry, as BrdU does. This constitutes a new argument for considering that the segmentation and the asymmetry of chromatids depend, at least partly, on two different mechanisms, where proteins are probably involved. Another effect of 5-ACR is to increase chromosome associations by satellites, secondary constrictions, and telomeric regions.


Studies on the altered electrophoretic type of the factor VIII related antigen. A distinct sub-group of von Willebrand's disease is characterized by an electrophoretically faster mobility of the factor VIII related antigen. Some of the physico-chemical properties of this variant antigen were investigated in this communication. The effect of temperature was tested by heating aliquots (0.5 ml) for 20 minutes at 45 degrees C, 56 degrees C and 65 degrees C. The variant was found to be denatured at 56 degrees C while the control was denatured at 65 degrees C. The effect of pH was tested by assessing the quantity (Laurell technique)  and electrophoretic mobility (two dimensional immunoelectrophoresis) of the antigen in a variety of buffers ranging in pH from 7.0 to 9.5. The quantity of antigen was variable both among variants and controls and the electrophoretic mobility of the variant antigen was faster at all pH's. Molecular weight differences between the variant and controls were not detected since the chromatographic profile of the variant was similar to that of the controls in Sepharose 6 B using a 0.02 M Tris-NaCL buffer at pH 7.0. The affinity of the antigen for human antibody was heterogeneous although the variant exhibited less affinity for one of the human antibodies but not the other. The inhibitory effect was more pronounced in serum than in plasma. Purified IGG, however, did not show any inhibition, as the residual antigen assayed by the Laurell technique, was similar to the expected values. This would imply that non-IgG plasmatic factors could also play a part in the observed inhibition.


Coal fires, industrial emissions and motor vehicles as sources of environmental carcinogens. One of the most widely studied carcinogenic agents in the environment is the polycyclic hydrocarbon, benzo(a) pyrene. As a component of soot from the inefficient combustion of coal, its association with cancer can be traced back 200 years, but its possible relevance to lung cancer as a widely distributed air relevance to lung cancer as a widely distributed air pollutant has been investigated only during the past 25 years. Domestic coal fires have been shown to be important sources, and smaller amounts come from industrial sources and from motor vehicles. There is evidence now that the concentration of benzo (a) pyrene in large towns in Britain has decreased by a factor of about ten during the last few decades, as a result of changing heating methods and smoke control. In view of the overwhelming effect of cigarette smoking, it is difficult to determine whether the benzo(a)pyrene content of the air has had any importnat effect on the development of lung cancer, but careful analysis of trends in mortality may now throw some light on this. Among other materials with carcinogenic properties that may be dispersed into the general air, asbestos is the one that has been investigated most thoroughly. The association between exposure to asbestos and the development of lung cancer and mesothelioma of the pleura has been clearly demonstrated among people occupationally exposed to the dust, but as far as the general public is concerned, any risk may be limited to the immediate vicinity of major sources. These and other hazards demonstrated among occupational gropus serve as a warning however to maintain careful scutiny of urban air pollutants in relation to the acetiology of cancer.


An in vitro model of canine immediate-type hypersensitivity reactions. Fragmented lung prepared from dogs cutaneously sensitive to ascaris antigen released histamine and a slow-reacting substance of anaphylaxis-like (SRS-A) material upon antigen challenge. Passive sensitization of fragments with serum obtained from ascaris-sensitive donor dogs enhanced the release of both substances; heating (56 degrees C for 4 h) destroyed the ability of the serum to enhance release. In passively sensitized tissues isoproterenol and papaverine inhibited the release of histamine and SRS-A; propranolol antagonized the effect of isoproterenol. Carbachol enhanced the release of both substances from passively sensitized lung wheras disodium cromoglycate and aminoguanidine were without effect. It is concluded that fragmented canine lung is a useful model for study of immediate-type hypersensitivity reactions.


Influence of Q- and G-banding on the Feulgen-stainability of human metaphase chromosomes. In this paper, model experiments on chicken red blood cell nuclei are described concerning the influence of methanol-acetic acid fixation and irradiation at different wavelengths, with and without prior Atebrin staining on subsequent Feulgen-stainability. In addition, data are reported on the influence on Feulgen-stainability of Giemsa-banding procedures, illumination of unstained chromosomes at 220 and 515 nm and exposure of unstained and Atebrin-stained chromosomes to illumination at 440 nm. The ASG and especially the trypsin-Giemsa technique appeared to reduce markedly Feulgen-stainability. The same holds true for Atebrin fluorescence of chromosomes. The data are discussed in relation to their implications for the assumed cause of the Q- and G-banding phenomena. Techniques are described that allow reliable Feulgen DNA measurements of individual chromosomes after application of either G- or Q-banding.


An ultrastructural study of osmiophilia in the human rectum. Rectal biopsies from subjects with a normal rectum and from patients with various forms of inflammatory bowel disease were studied by the prolonged osmication technique. No consistent ultrastructural differences were observed between these groups, but there were striking differences between individual epithelial cells in the same biopsy and between the epithelium and the cells of the lamina propria. The Golgi apparatus was demonstrated occasionally in the epithelium, often in endothelium. Endoplasmic reticulum, perinuclear cisterna and mitochondria were variably outlined. In plasma cells, there were striking differences in osmiophilia. The underlying mechanism of the different staining patterns is not clear. The findings do not appear to help in the differential diagnosis of inflammatory bowel disease nor to shed any new light on their underlying pathogenic mechanisms.


Histochemical detection of sialic acid residues using periodate oxidation. The use of low concentrations of periodate for the detection of sialic acid residues in tissue sections has been investigated. Oxidation of aqueous solutions of sugar glycosides with 0.4 mM periodate revealed that sialic acid was oxidized more rapidly than other sugars found in glycoproteins. Sequential treatment of tissue sections with 0.4 mM periodate for 30 min followed by Schiff's reagent stained sialic acid residues but other sugar components were not stained under these conditions.


A comparison of polychromasia and reticulocyte counts in assessing erythrocytic regenerative response in the cat. Three staining techniques were evaluated for use in assessing the erythrocytic regenerative responses in the cat. Using new methylene blue as a vital stain, the aggregate reticulocyte count closely corresponded to the reticulocyte count on air-dried smears stained with new methylene blue and to the polychromatophilic erythrocyte count on Wright's stained smears.


Relationship between changes in the translational apparatus and actinomycin production in Streptomyces antibioticus. As previously reported (G. H. Jones, 1975), transfer ribonucleic acids (tRNA's) and ribosomes from actinomycin-producing cultures of Streptomyces antibioticus show a decreased ability to function in aminoacylation and translation as compared with the corresponding components from younger cells. Further, specific changes in the isoacceptor patterns are revealed when tRNA's from actinomycin-producing cells are compared with those of younger cells by reverse- phase column chromatography. A specific glycyl-tRNA species is eliminated from the reverse-phase profile of tRNA's from actinomycin-producing S. antibioticus cells as compared with younger cells. Changes in isoacceptor patterns were also observed for the amino acids methionine, valine, phenylalanine, and leucine. Actinomycin synthesis was inhibited by growing S. antibioticus cells in the presence of alpha-methyl-DL-tryptophan. Inhibition of actinomycin synthesis reversed the changes in tRNA observed in normally grown control cultures, although it had no demonstrable effect on the growth of the cells. Thus, tRNA from 48-h-old, alpha-methyl-tryptophan-grown cells had amino acid acceptor activity that was equal to or greater than that of tRNA from 12-h-old, normally grown cells. Similarly, the reverse-phase chromatographic pattern for glycyl-tRNA's from 48-h-old, alpha-methyl-tryptophan-grown cells was identical to that of the glycyl-tRNA's from 12-h-old, normally grown cells. In contrast, the ability of ribosomes from 48-h-old, alpha-methyl-tryptophan-grown cells to function in polypeptide synthesis in vitro was essentially identical to that of 48-h-old, normally grown cells. Ribosomes from 12-h-old, normally grown cells were severalfold more active in in vitro polypeptide synthesis.


The ovalbumin gene. In vitro enzymatic synthesis and characterization. Using purified single-stranded ovalbumin complementary DNA (cDNAov) as a template for avian myeloblastosis (AM) virus reverse transcriptase, we have enzymatically synthesized a complete double-stranded cDNAov sequence. Our data suggests that many single-stranded cDNAov molecules contain short double-stranded sequences (hairpins) at their 3' termini capable of acting as primers for synthesis of complete double-stranded cDNAs. Optimum conditions for synthesis of the double-stranded cDNAov were found to be a high temperature (46 degrees) and a low salt concentration. Nevertheless, in all cases 40% of the initial single-stranded cDNAov molecules fail to prime for synthesis of a complementary double strand. Following synthesis, the second DNA strand is covalently linked to the first cDNAov strand as shown by analysis on alkaline sucrose gradients. The two strands have a high Tm on hydroxylapatite (89 degrees). These intact double-stranded cDNAov structures have a bouyant density in CsCl gradients of 1.700 g/cm3 and rapidly renature after heat denaturation with a C0t1/2 value of less than 2 X 10(-6) mol s liter(-1). All size classes of cDNAs, i.e. partial as well as complete transcripts of the mRNA, are capable of forming double-stranded structures. The closed loop of the double-stranded cDNAov could be opened with S1 nuclease. The denatured complementary strands of the cDNAov then renatured with the appropriate second order kinetics at a C0t1/2 value of 1.89 X 10(-3) mol s liter(-1). Using the enzyme terminal deoxyribonucleotidyltransferase to label to free 3'-terminal end of double-stranded [32P]cDNAov with 3H, we demonstrate a convenient procedure to study the site for restriction endonuclease cleavage within the ovalbumin gene.


Light-induced exchange of nucleotides into coupling factor 1 in spinach chloroplast thylakoids. The method of centrifugation of chloroplast thylakoids through silicone fluid, previously used to estimate the uptake of solutes by thylakoids, is shown to be an excellent method for measuring binding of nucleotides to thylakoids. This binding, which is probably an exchange (Harris, D. A. and Slater, E. C. (1975) Biochim. Biophys. Acta 387, 335-348), is enhanced by light and is sensitive to uncoupling. Half-maximal binding of adenosine 5'-triphosphate (ATP) or adenosine 5'-diphosphate (ADP) at 10 mjM was reached within less than 0.1 s. With illumination times sufficient to elicit maximal binding, saturation of the site(s) is approached at 20 muM nucleotide and dissociation constants of 5 muM and 7 muM were calculated for ADP and ATP, respectively. At saturation, the binding corresponds to 1 mol/mol of coupling factor 1 or less. Although the light-dependent binding of ADP does not require Mg2+, that of ATP is markedly enhanced by Mg2+. A 10-fold molar excess of guanosine di- or triphosphate or adenyl-5'-yl imidodiphosphate had little effect on the binding. Adenosine 5'-phosphosulfate, a competitive inhibitor of phosphorylation with respect to ADP, decreases the binding. Thylakoids, previously illuminated in the absence of added nucleotides, retain the capacity to bind ADP or ATP in the dark long after the H+ electrochemical gradient has decayed. The conformation of coupling factor 1 in darkened thylakoids following illumination in the absence of added nucleotides may thus differ from that in thylakoids either illuminated in the presence of nucleotides or kept in the dark. Approximately 20% of the ADP bound to coupling factor 1 in thylakoids is converted to ATP by a 2-s illumination. Bound inorganic phosphate, derived either from ATP or from inorganic phosphate itself, serves as the phosphoryl donor. Bound ADP may, therefore, be of catalytic significance in the mechanism of phosphorylation.


Treatment of pathological fractures of long bones excluding those due to breast cancer. Seventy-two pathological fractures associated with tumors other than carcinoma of the breast in the long bones of the extremities of sixty patients were treated over a five-year period at Roswell Park Memorial Institute. Pain was relieved in 91 per cent of the patients treated by internal fixation, in 59 per cent of those treated by irradiation, and in 45 per cent of those treated by other means. Among patients with lower-extremity fractures, 61 per cent of those treated by internal fixation became ambulatory, whereas only 23 per cent of those treated by other methods were able to walk. Internal fixation of these pathological fractures appeared to be the best treatment.


Resolution in electron microscope autoradiography. III. Iodine-125, the effect of heavy metal staining, and a reassessment of critical parameters. Resolution for 125I-labeled specimens under electron microscope (EM) autoradiographic conditions was assessed experimentally. With this isotope the size of the silver halide crystal was the most important resolution-limiting factor. Heavy metal staining such as is routinely used in preparing animal tissues for EM autoradiography produced an improvement in resolution of approximately 15-20%. For a 500-1,000-A biological tissue section fixed with OsO4 and stained with uranyl acetate, we obtained resolution (half distance, HD) values of approximately 800 +/- 120 A using Ilford L4 emulsion and 500 +/- 70 A using a Kodak NTE-type emulsion. General aspects of resolution-limiting factors and comparison with 3H and 14C values are discussed.


The electrophysiological mapping of compartments within a mammalian cell. The electrical properties of structures within an intact cell were examined by impalement with micropipette electrodes. Mean potential differences (PDs) measured from interphase HeLa cells showed that internal membrane-bounded compartments such as the nucleus, Golgi region, and the mitochondria were more negative than the cytoplasm with respect to an external grounding electrode. The nuclear PDs, unlike Golgi and cytoplasmic PDs, shifted during interphase and reached a peak value shortly before mitosis. The positioning of micropipettes was confirmed by electron microscope examination of marker solutions that were microinjected into specific intracellular regions. The combination of methods described here offers a new approach for the study of physiological events within intact, living cells.


Regular structures in membranes: the lumenal plasma membrane of the cow urinary bladder. The ultrastructure of the lumenal plasma membrane of the cow urianry bladder has been studied in thin sections of glutaraldehyde- and glutaraldehyde-H2O2-fixed specimens, by negative staining and freeze fracture. A regular hexagonal array of particles confined to polygonal plaques 0-1-0-4-mum in diameter and separated by 0-02-mum interplaque areas is revealed by all 3 techniques. Cross-sections through particulate areas fixed with glutarayldehyde-H2O2 display a tetralaminar structure consisting of the usual approximately 8-nm-thick trilamellar unit membrane structure, on the external dense leaflet of which is located an additional approximately 4-nm-thick stratum which is occasionally resolved into a row of regulrly spaced approximately 4-nm-diameter particles. Non-particulate areas feature only the approximately 8-nm-thick trilamellar structure. Tangential sections reveal an hexagonal array of particles with a unit cell of approximately 16 nm. Four membrane faces can be revealed by freeze fracture and etching of membranes of the cow urinary bladder; 2 complementary split inner membrane faces (A and B) revealed by the cleaving process and the lumenal and cytoplasmic membrane surfaces exposed by etching. Face B, which belongs to the external membrane leaflet and faces the cytoplasm, displays plaques of particles arranged in a hexagonal lattice with a unit cell of approximately 16 nm. Face A, which belongs to the cytoplasmic membrane leaflet and faces the lumen, displays a complementary array of hexagonally packed pits. The hexagonally arranged particles also protrude into the lumenal membrane surface where they can occasionally be resolved into 6 approximately 5-nm-diameter subunits; the cytoplasmic surface appears smooth. Six approximately 5-nm-diameter subunits are also revealed in negatively stained preparations. The data are consistent with a model for the membrane in which the particles forming the hexagonal structure protrude above the lumenal membrane surface and also bridge most of the thickness of the membrane.


Interrelationships between Golgi, GERL and synaptic vesicles in the nerve cells of insect and gastropod ganglia. In addition to demonstrating synaptic vesicles, staining with the zinc-iodide-osmium tetroxide (ZIO) method reveals the presence of positively reacting GERL membranes in association with the Golgi complex and lysosomes in the nerve cell bodies within ganglia from the locust Schistocerca gregaria and the gastropod molluscs, Limnaea stagnalis and Helix aspersa. A positive response to ZIO occurs in certain Golgi vesicles and saccules, in GERL (Golgi-endoplasmic-reticulum-lysosomes), in multivesicular bodies as well as residual bodies and in small vesicles and cisternae of axonal smooth endoplasmic reticllum (ER). The interrelationships between these organelles are considered in view of the similarity of the ZIO localization to phosphatase-rich sites in the neuronal perikarya and with respect to the possibility that components of the synaptic vesicles are formed in the Golgi region of the cell and migrate via the axonal smooth ER to the synaptic regions.


Thin-layer chromatography of some cationic dyes commonly used in histology. A number of cationic dyes commonly used in histology may be conveniently and effectively analysed using the thin-layer chromatographic system of Marshall and Lewis. Commercial samples of Acridine Orange, Crystal Violet, Janus Green B,Methyl Violet, Neutral Red, Pyronin B, Pyronin Y(G), Safranin, Victoria Blue B and Victoria Blue 4R have been analysed with this system. All have been found to be complex mixtures of coloured components. Chromatographic data on these components are presented.


Androgens in patients with benign prostatic hyperplasia before and after prostatectomy. Plasma androgens [testosterone (T), 17beta-hydroxy-5alpha-androstan-3-one (DHT), androst-4-en-3,17 dione(A), and dehydroepiandrosterone (DHEA)] as well as 17 hydroxyprogesterone were measured in a group of patients (age 60-80 yrs.) with benign prostatic hyperplasia (BPH) just before prostatectomy and compared to values obtained in subjects of similar age without signs of BPH. The most important difference was observed in the mean DHT level which was significantly (P less than 0.025) higher than in the control group; mean T and free testosterone levels in BPH patients were slightly higher (P less than 0.05) in the age group 70-80 yrs; whereas in age group 60-70 mean values were similar to those observed in normal controls. Mean A, DHEA and 17 OHP and E2 levels were not significantly different in BPH patients when compared to age matched controls. 2-5 months after prostatectomy, T and DHT levels were significantly higher than immediately preoperatively. The preoperative stress may have influenced the preprostatectomy values.


Delta5-androstenediol: kinetics of metabolism and binding to plasma proteins in normal men and women. Using the constant fusion and single injection technique the metabolic clearance rates (mean +/- SEM) for delta5-androstene-3beta, 17beta-diol (delta5-idol) were measured for 19 normal men (1311 +/- 67 1/24 h) and 10 normal women (858 +/- 63 1/24 h). The constant infusion technique yielded values for the conversation ratios for the transformation of delta5-diol to several products: dehydroepiandrosterone (DHEA)/delta5-diol of 0.06+/-0.01 for men and 0.05 +/- 0.01 for women, of delta5-diol sulfate/delta5-diol of 0.45 +/- 0.04 for men and 0.52 +/- 0.03 for women and of DHEA sulfate/delta5-diol of 5.53 +/- 0.26 for men and 5.02 +/- 0.42 for women. The single injection technique yielded rate constants (units) and volumes of distribution (liters) for delta5-diol; Ki = 34.3 +/- 4.3 for men and 35.0 +/- 3.9 for women, K2 = 63.7 +/- 4.1 for men and 75.1 +/- 4.2 for women, V1 = 23.1 +/- 3.2 for men and 11.9 +/- 2.3 for women, V2 = 14.8 +/- 3.7 for men and 9.2 +/- 3.2 for women. The mean delta5-diol plasma concentration was 1.08 +/- 0.10 ng/ml for 12 men and 1.17 +/- 0.16 ng/ml for 9 women. (he calculated blood production rates for delta5-diol were 1357 +/- 117 mug/24 h for 12 men and 969 +/- 131 mug/24 h for 9 women. The per cent binding (equilibrium dialysis) was higher for women (94.9 +/- 0.3) than for men (93.0 +/- 0.2). Paper electrophoresis showed that significant fractions of 3H-delta5-diol migrated with both the beta-globulin and albumin fractions. Estrogen administration to two normal men increased the per cent binding of delta5-diol to plasma proteins and decreased the metabolic clearance rate towards the values found for normal women.


Interhemispheric neocortical connections of the corpus callosum in the reeler mutant mouse: a study based on anterograde and retrograde methods. The tangential organization of the callosal system of interhemispheric connections, as judged by the distribution of axon terminals as well as by the distribution of cells of origin of callosal axons, is normal in the reeler mutant mouse. As in the normal animal connections between the two cerebral hemispheres are homotopic. In the reeler, as in the normal animal, medium-sized pyramidal cells are, numerically speaking, the principal cells of origin of the callosal system. These lie superficially in the cortex of the normal animal but deep within the cortex of reeler. Callosal terminals are most densely concentrated at the cortical level of the small and medium-sized pyramids in both reeler and normal animals. It is probable, therefore, that the same classes of neurons are interconnected by the callosal system in the normal and reeler mouse despite malposition of neurons in reeler. The patterns of intracortical distribution of terminals of callosal axons is evidently governed by the positions of their target cells.


The anatomical organization of the cerebello-olivary projection in the cat. The cerebello-olivary pathway in the cat has been examined using orthograde and retrograde neuroanatomical tracing techniques. The orthograde transport of 3H-leucine from injection sites in the deep cerebellar nuclei labeled dentate and interpositus projections to the rostral two-thirds of the contralateral inferior olivary complex. These projections are topographically organized, with the dentate nucleus projecting to the principal olivary nucleus and the posterior and anterior interpositus nuclei projecting to the medial and dorsal accessory olives respectively. Fibers from the ventral half of the dentate nucleus terminate in the lateral bend and ventral lamina of the principal olive, whereas the medial and lateral parts of the dorsal half of the nucleus project to the medial and lateral regions of the dorsal lamina respectively. It is apparent that the more caudal parts of the interpositus nuclei project to areas of the medial and dorsal accessory olives near the caudal end of the principal olivary nucleus, whereas neurons in the more rostral parts of the interpositus nuclei project to the more rostral areas of the accessory olivary nuclei. A connection between the fastigial ncleus and the inferior olive could not be demonstrated. The retrograde transport of horseradish peroxidase (HRP) from injections sites in the inferior olive labeled cells throughout the contralateral dentate and interpositus nuclei. The labeled cells were especially numerous in the ventral parts of the dentate and posterior interpositus nlclei. These HRP-positive neurons were consistently small (10--15 mu) ovoid or spindle-shaped cells, with relatively large nuclei and light-staining Nissl substance. This evidence strongly suggests that the cerebello-olivary pathway originates from a population of small neurons in the dentate and interpositus nuclei and projects to specific, topographically defined areas in the contralateral inferior olive.


Epidermal changes in lichen planus.

Epidermal changes in lichen planus. Ultrastructural investigations in 15 patients with lichen planus (LP) revealed a multitude of epidermal changes. The most consistent findings were (1) multiplication, irregular folding or dislocation of the basal lamina, (2) fragmentation with degenerative changes of basal keratinocytes, (3) formation of numerous fibrillar Civatte bodies and, (4) presence of dyskeratotic elements. In addition, mitotic figures of keratinocytes and Langerhans cells were observed. These findings support the view that the primary event in LP represents an injury involving epidermal basal cells. Furthermore, the replacement of destroyed tissue seems to originate not only from the margin of the lesions or the skin appendages but also from unaffected cells within the LP papule.


Determination and identification of polycyclic aromatic hydrocarbons in smoked and charcoal-broiled food products by high pressure liquid chromatography and gas chromatography. A high pressure liquid chromatographic procedure has been developed and applied to the analysis of polycyclic aromatic hydrocarbons (PAH's) in 70 samples of smoked food products commercially available in Canada, and in 6 charcoal broiled meats. In some cases a gas-liquid chromatographic procedure was used as a confirmatory technique. In the commercial samples PAH's were detected in approx. 70% of the samples. Total PAH's ranged from 0-141 ppb and individual PAH's from 0-38 ppb. With the charcoal-broiled samples, total PAH's and individual PAH's ranged from 0-164 ppb and 0-60 ppb respectively. These levels are similar to those observed in other countries.


[Alpha-1-antitrypsin, orosomucoid, transferrin and alpha fetoprotein in the amniotic fluid, Changes in concentration in normal and pathologic pregnancies]. In this article, which is a preliminary report, the authors determined amniotic fluid levels of alpha-I-antitrypsin, orosomucoid, transferrin and alpha-fetoprotein during complicated pregnancies with gestational ages of 31 to 42 weeks. These levels were compared with those observed during normal pregnancies. The clinical value of the variations of the glycoprotein levels is discussed.


Use of actinomycin D for the specific quenching of fluorescence of deoxyribonucleic acid in cells stained with acridine aminoderivatives. Actinomycin D specifically quenches the fluorescence of acridine orange and quinacrine bound to deoxyribonucleic acid in cytologic preparations, but does not change the fluorescence of these fluorochromes bound to RNA. The following fluorescence-cytochemical applications of techniques based on these findings can be suggested: (a) distinction between deoxyribonucleic acid and ribonucleic acid; (b) detection of double-stranded virus ribonucleic acid; (c) approximate estimation of the lengths of A-T sequences in deoxyribonucleic acid molecules.


Immunohistochemical localization of melatonin in the rat Harderian gland. Using fluorescence and double antibody techniques, melatonin was localized immunohistologically in the secretory cells of the Harderian gland of mature male rats. The presence and quantity of melatonin in the acinar cells seem to correlate with the amount of porphyrins inside the lumen. The specificity was proven by disappearance of yellow fluorescence after saturation of antibody with melatonin or after use of nonspecific antibody only.