Synergistic effects of the combination of cis-platinum diamminodichloride and 2,2'-anhydro-1-beta-D-arabinofuranosyl-5-fluorocytosine in transplanted mouse leukemias. cis-Platinum diamminodichloride has been studied in combination with 2,2'-anhydro-1-beta-D-arabinofuranosyl-5-fluorocytosine on an every-4-day schedule in various lines of mouse leukemia. This combination is synergistic in leukemias L1210 and P388 and sublines made resistant to 5-fluorouracil or methotrexate. There is no cross-resistance between cis-platinum diamminodichloride and 2,2'-anhydro-1-beta-D-arabinofuranosyl-5-fluorocytosine, but the combination is no more effective against lines of leukemia made resistant to cis-platinum diamminodichloride or to 2,2'-anhydro-1-beta-D-arabinofuranosyl-5-fluorocytosine than either single active compound alone. Since these compounds have no cross-resistance, act by quite different mechanisms of action, and have different limiting toxicity, the combination is now being evaluated clinically.


Chemotherapy of non-Hodgkin's lymphoma: 10 years' experience in the Southwest Oncology Group. The Southwest Oncology Group (SWOG) has completed five studies of high-dose intermittent combination chemotherapy for the management of advanced (stage III and IV) non-Hodgkin's lymphoma involving 1143 patients from May 1966 to September 1974. Lack of uniform histopathologic interpretation precludes precise analysis of these data. Although there has been little change in complete response duration over the years of this study, there has been an overall improvement in response rate and survival though there is no statistically significant improvement in the best overall survival when compared to the Stanford experience in stage III and IV disease (1960-71). The response rate and survival in diffuse histiocytic lymphoma have improved since the first study. There is definite evidence of a plateau in the survival curve beyond 2 years. The percentage of survival at which the plateau appears has increased over the years to 40% in the most recent studies, and the survival is suggestively better than the Stanford experience (P = 0.09). Over the years there has been a distinct improvement in response rate and survival of patients with nodular lymphocytic lymphoma, although the best SWOG survival is no different than the Stanford experience (P = 0.36).


Chemotherapy of non-Hodgkin's lymphomas: Eastern Cooperative Oncology Group experience. The addition of prednisone to the two-drug combination of cyclophosphamide and vincristine increases the number of complete remissions (CR). Patients who had CRs in at least two of the studies, EST 0168 and EST 3472, lived significantly longer than those who had partial remissions. In EST 0168, the survival time correlated strongly with the nodal pattern and cell type, confirming results of Rosenberg and co-workers. EST 1472 confirmed the activity of BCNU in lymphocytic lymphomas; when both agents are combined with prednisone, the induction potential of BCNU is equal to that of cyclophosphamide. In the histiocytic lymphomas, BCNU added to CVP increases the percentage of CRs but it does so at the expense of considerable toxicity. EST 0871 demonstrated that low-dose bleomycin and hexamethylmelamine are active agents in lymphoma and should be combined with other effective combinations in an attempt to improve the CR rate. Finally, our current studies demonstrate that the failure rate in the diffuse or unfavorable lymphomas is higher than that in the more favorable nodular forms.


Overview of four clinical studies of chemotherapy for stage III and IV non-Hodgkin's lymphomas by the Cancer and Leukemia Group B. This paper presents an overview of four Cancer and Leukemia Group B studies in 1266 patients with stage III-IV non-Hodgkin's lymphoma. The cases were analyzed across protocols; the major prognostic determinants were prior chemotherapy, age, and histology. The four studies proved that cyclophosphamide maintenance was superior to no maintenance even after prolonged intensive induction chemotherapy. Furthermore, the reinforcement program of monthly pulse doses of vincristine and prednisone, whose value was established in the treatment of acute leukemia, led to highly significant improvement in remission duration and survival. Other facets of the chemotherapy programs are still being subjected to analysis, but this report sets out some preliminary conclusions.


Non-cross-resistant combinations in stage IV non-Hodgkin's lymphomas. Two triple-drug combinations (cyclophosphamide, vincristine, and prednisone [CVP] and adriamycin, bleomycin, and prednisone [ABP]) were randomly tested in 57 consecutive patients with pathologic stage IV non-Hodgkin's lymphoma. No statistical difference was observed in the incidence of complete remission (CR) (48% vs 50%), median duration of CR (10.5 vs 20.5 months), and survival. CR correlated positively with survival. After crossover for progression or relapse, no cross resistance was detected between the two combinations (CR plus PR = 40% for CVP and 50% for ABP). In a subsequent prospective study CVP and ABP were sequentially alternated in an attempt to improve the incidence and duration of CR. In the first 32 evaluable patients CR was produced in 50%. The findings are still preliminary.


Team approach to management of non-Hodgkin's lymphomas: past and present. A detailed review of our results of radiotherapy for stage I and II non-Hodgkin's lymphomas has been presented as background for our current multidisciplinary approach which uses multiple-agent chemotherapy, radiotherapy, and immunotherapy for nodular and diffuse stage I, II, and III lymphomas of the poorly differnetiated lymphocytic, mixed, and histiocytic varieties. Optimum management is based on coordinated efforts of medicine, radiotherapy, pathology, diagnostic radiology, and surgery. In stage I, II, and III disease, routine staging procedures include lymphangiography, bilateral iliac crest needle biopsies of the bone marrow, and percutaneous needle biopsies of the liver. Staging laparotomy is considered essential to our present program for stage I, IE, II and IIE disease. In our opinion, staging laparotomy is not indicated in stage II or IIIE disease. Although very preliminary, results for these programs are encouraging. This paper presents the concept of team management and a specific plan for its implementation.


[Detection of a minor stimulating product involved in secondary allogenic proliferation of human lymphocytes in vitro]. In a family with a shared parental haplotype studied in MLR I and II we report that: 1) A secondary proliferation can be induced without a primary positive MLR; 2) In these conditions a minor determinant activating secondary proliferation is detected; 3) No significant association of this product with the available makers (HLA-A, B, C, D, Ly-Li) of the HLA region has been found so far; its localisation within or outside the MHC is under investigation.


[Evidence of protein kinase activity in 2 murine oncornaviruses]. A protein kinase activity has been detected in two strains of murine Oncornaviruses, MSV/MLV and EFV. This activity phosphorylates not only endogenous viral proteins but also exogenous substrates (histones and phosvitin). The stimulation of enzyme activity by detergents along with the increase of specific activity in viruses treated with trypsin during purification suggest that the enzyme is located in the viral particle.


The enlargement of small pulmonary arteries by preliminary palliative operations. Four patients with tetralogy of Fallot, three of whom had congenital pulmonary atresia, were treated by initial palliative operations to enlarge left and right pulmonary arteries which were considered too small for complete repair. Two to four years later the right and left pulmonary arteries had enlarged sufficiently to allow complete repair.


Neurosurgical control of pain in the patient with cancer. If properly evaluated, the majority of patients with intractable pain caused by visceral as well as somatic malignancy can obtain relief. Analgesics are of great value in some cases. They are especially indicated for patients who have conditions producing pain of short duration or for terminal patients. Radiotherapy may temporarily help most patients with intractable pain. Open or stereotactic surgical intervention and especially interruption of the ascending pain fibers within the spinal cord may give permanent relief to patients who have increasingly severe pain. At present the best results are obtained with percutaneous high cervical electrocoagulation of the spinothalamic tracts. Experience with electrical inhibition of pain based on the gate theory is limited. In the future, however, it may replace the present destructive approaches.


Specific break points in chromosomally abnormal human fibroblast subpopulations. Common chromosome break points were found in a banding study of 25 chromosomally abnormal fibroblast clones derived from normal individuals and patients exposed to environmental carcinogens. Of 81 break points, 38% appeared to be in terminal bands. This specificity did not appear to be the consequence of the biased recognition of exchanges. In addition, specificities for break points at exact interstitial loci were detected. The presence of clones in cultures grown in a wide variety of conditions, the finding of two identical clones in two independent lines from the same biopsy, and the absence of any obvious continuing instability in lines support the view that chromosomally abnormal fibroblasts occur in vivo.


Antepartum evaluation of the high-risk fetus: problems and prospects. The chief obstetrical problems encountered today in the prenatal evaluation of the high-risk fetus are presented. Advantages and pitfalls or recent techniques utilized in the management of the high-risk pregnancy are discussed. They include: a prenatal scoring system for identifying the high-risk population; examination of the karyotype of cells in amniotic fluid, and quantitation of alpha-fetoprotein levels in maternal plasma and amniotic fluid for the early prenatal detection of birth defects; ultrasonography for the intrauterine diagnosis of fetal growth retardation and assessment of fetal maturity; the use of maternal urinary estriol excretion, maternal plasma human placental lactogen levels and the oxytocin stress test for the early detection of fetal distress; estimation of fetal maturity by amniotic fluid analysis of lecithin or lecithin-sphingomyelin ratios, creatinine and Blue Nile fetal cell staining. Newer, still experimental, techniques (e.g., fatal breathing movements, fetoscopy, and dehydroepiandrosterone plasma clearance) are viewed in light of further possible decreases in maternal and perinatal mortality.


Critical periods for behavioral anomalies in mice. While mice have been used less frequently than rats in behavioral research, there use has some advantages in teratological studies. The development of the mouse CNS has been investigated more extensively than that of the rat. Since time of insult has been found to be an important factor in effects on both anatomy and behavior, data on the sequence of events in CNS development are valuable in planning and interpreting behavioral assessments of potential teratogens. A comparison of studies in mice and rats suggests that behavioral effects of teratogens are similar in the two species and demonstrates that mice can be used successfully in a variety of behavioral evaluations.


Environmental transport and transformation of automotive-emitted lead. The use of a mass balance equation assists in clarifying the contribution of automotive-emitted lead to the regional and global problems of the distribution of this element, by taking into account the physicochemical transformation processes operating on the lead during its transport along the various environmental pathways. The technical factors which govern the emission of such lead from automobile engines and which affect its subsequent dispersion through the environment are described.


Emission control devices, fuel additive, and fuel composition changes. Emission control devices are installed to meet the exhaust standards of the Clean Air Act for carbon monoxide and hydrocarbons, and it is necessary to know, from a public health point of view, how exhaust emissions may be affected by changes in fuel additives and fuel composition. Since these topics are concerned with developing technologies, the available literature on exhaust emission characteristics and the limited information on health effects, is reviewed.


Frequency distribution of messenger sequences within polysomal mRNA and nuclear RNA from rat liver. DNA complementary to polysomal poly(A)-containing mRNA (cDNA) of male rat liver was used to study the diversity of messenger sequences in the nucleus and in polysomes. 1. Hybridization of cDNA against an excess of its own polysomal mRNA template revealed that about 10,000 different mRNA species are expressed in the liver tissue. They are distributed in a wide frequency range derived from approximately 0.5% of the total genome. 2. Hybridization of the cDNA against total nuclear RNA shows that messenger sequences comprise less than 1% of the mass of total nuclear RNA. Messenger sequences have a different frequency distribution in nucleus and cytoplasm. 3. In hybridizations using cDNA, which had been fractionated into sequences representing abundant and scarce polysomal mRNA molecules, it was found that although abundant cytoplasmic messenger sequences are also abundant in the nucleus, they exist in a significantly lower frequency range in the nuclear compartment.


Recording of drug prescriptions in the county of Jämtland, Sweden. III. Drugs presented for blood donors in a 5 year period. Drug prescriptions in the county of Jämtland, Sweden, have been examined in order to estimate drug exposure of blood donors. During the period 1970-74 a donor group (n = 292) purchased significantly more drugs than non-donors matched for age, sex and residential area in the county. This was true for most drug categories, except for cardiovascular and endocrine drugs and iron preparations. The actual consumption of the drugs obtained was not investigated. Prospective studies are required to assess the clinical consequences of the surprisingly high drug exposure of blood donors.


The private specificity H-2.4 and the public specificity H-2.28 of the D region are expressed on two independent polypeptide chains. The antigenic specificities H-2.4 (private) and H-2.28 (public) in the H-2a haplotype are controlled by the D region of H-2 as defined by the available recombinants. In previous studies we have demonstrated by the antibody-induced redistribution method that the antisera against these specificities contain antibodies against at least two different polypeptide chains. We here report the results of the indirect immunoprecipitation of radiolabeled antigens after solubilization with Nonidet-P40. The antisera against the two specificities precipitated from such extracts two different and independent polypeptide chains, indicating that the products of the D region, as presently defined, comprise at least two different molecules. The molecular weight of both chains is approximately 45 000, which is similar to other molecules bearing private H-2 antigenic specificities. Consequently, the chromosomal segment presently defined by recombination studies as the D region, must contain another locus, controlling the second polypeptide chain which is detectable by anti-H-2.28 antisera, besides the H-2D locus which controls the polypeptide chain bearing the private specificity H-2.4 as well as most of the public specificities.


Procion yellow staining of motoneurones in the frog. Intracellular recording and subsequent staining of spinal motoneurones in the frog was made by procion-filled micropipettes. Spike discharges in response to dorsal root (DR) and ventral root (VR) volleys as well as to direct current injections were studied. Reconstruction of the dendritic tree of the cell stained after recording was made from photomicrographs taken from frozen serial sections of the spinal cord. Migration of the dye into a neighbouring unimpaled cell was observed. The advantages of the procion injection technique in studying the frog's spinal cord physiology are discussed.


Electron microscopic investigation on the zona glomerulosa of rats after treatment with heparin and potassium. Adrenal zona glomerulosa of rats was investigated by means of electron microscopy after stimulation with potassium and suppression with heparin. Both intreated and control animals two types of mitochondria -- one with tubular and the other with tubulosaccular cristae -- were found. Stimulation with potassium increased the amount of mitochondria with tubular cristae, heparin treatment caused a simplification of the structure of tubulosaccular cristae. It was assumed that these two mitochondrium-poulations contain different enzyme systems, which catalyze the different step-series of aldosterone biosynthesis. On the basis of cytoplasmic density, dark and light cells could be distinguished only in the zona glomerulosa of control rats, they were absent in both groups of treated animals. The density of these cells probably did not depend on their metabolic activity, but rather on their hormone storage.


Activity of acid: Co A ligase (AMP) in the course of experimental allergic encephalomyelitis. In studies concerned with the activation of fatty acids, a metabolic step initiating the biosynthesis of fatty acids rendering them suitable for elongation, desaturation, esterification or degradation, we have demonstrated that the activity of acid: Co A ligase (AMP) -- E.C.6.2.1.3. is declined throughout the whole investigated period of experimental allergic encephalomyelitis (EAE). This observation clearly indicates that in this experimental disease it is not only the increased catabolism of lipid constituents furnishing the various membraneous structures, including myelin sheats, but also an overall impaired capacity of synthesing new lipids by the diseased cells of the nervous system which must be taken into consideration.


Continuous lymphoblastoid suspension cultures from cells of haematopoietic organs of baboons with malignant lymphoma. Report III: Immunological studies. Lymphoblastoid cultures established from haematopoietic organs of baboons with lymphoma belong to the B-cell type. Cytoplasmic antigen is revealed in 2--6% of suspension culture cells established from lymphomatous baboons by indirect immunofluorescence reaction using serum from human Burkitt's lymphoma patients with antibody titer to capsid antigen of Epstein-Barr virus 1: 320. There is a correlation between the number of antigen-containing cells in these cultures and the number of cells in which herpes virus is revealed electron microscopically. Sera of baboons with lymphoma contain high titers of antibodies to Epstein-Barr (EBV) virus. Specific differences between antigens being produced in cultures by HBV (herpes virus of baboon) and EBV (Epstein-Barr virus) are minimal, that indicates their immunologic similarity.


A quantitative study of encephalitogenic protein and peptides in guinea pigs. The encephalitogenic activity of bovine encephalitogenic protein (EP) at at doses from 1 to 33.5 microgram was studied in guinea-pigs. Anencephalitogenicity can be demonstrated at a dose of 1 microgram of EP when given with Freund's complete adjuvant. An increase in the dose of EP results in an increase in encephalitogenicity. Data from a previous paper (Lindh and Bergstrand 1975) supplemented with new data on the encephalitogenic capacity of peptide 89--169 were compared concerning the incidence and strength of experimental allergic encephalomyelitis (EAE) to determine the relative contribution of the various parts of EP to the encephalitogenic properties of the total molecule. Peptide 89--169 shows the same degree of encephalitogenicity as EP, but when the tryptophan residue is blocked (HNB-89-169) the encephalitogenicity is reduced to approx. 15% of the initial amount. Peptide 43--115, tyr (modified at the tyrosine residue 67) appears to have an encephalitogenicity of rather less than 10% of the intact EP molecule, whereas peptide 1--42, if active at all, has an encephalitogenicity of approx. 1% of the total EP molecule. The findings are discussed in the light of a possible cross-reactivity between different determinants of EP.


Identification of rho-antigenic determinants on the surface of mouse T-lymphocytes. A monospecific antiserum has been prepared in rabbits against purified rho-antigen, a 100,000 mol.wt glycoprotein found on the surface of mouse L-cells. This antiserum has been employed to demonstrate the presence of rho-antigenic determinants selectively on the surface of mouse T-, but not B-lymphocytes.


High inhibitory activity of R 5020, a pure progestin, at the hypothalamic-adenohypophyseal level on gonadotropin secretion. Synthetic "progestins" currently used in the contraceptive pill inhibited the luteinizing hormone (LH) responsiveness to LH-releasing hormone in cells in culture in a way undistinguishable from that of androgens. Moreover, they competed for binding of the 3H-labeled androgen R 1881 to the rat prostate androgen receptor and stimulated seminal vesicle and prostate weight in castrated rats. R 5020, a pure progestin, was without effect on the above-mentioned parameters. However, a complete inhibition of the LH surge measured in the afternoon of expected proestrus was obtained at a dose of R 5020 similar to that of D-norgestrel. The synthetic progestin was also found to inhibit ovulation and to delay vaginal cornification. The present data show that the synthetic "progestins" commonly used in the pill possess intrinsic androgenic activity which could well be responsible, to an unknown extent, for their effectiveness as contraceptive agents. R 5020, a synthetic progestin devoid of androgenic activity, is at least as potent as the most potent 19-nortestosterone derivative, D-norgestrel, in inhibiting gonadotropin secretion and other parameters of the estrous cycle in the rat. The availability of a pure progestin devoid of androgenic activity but highly effective as an inhibitor of gonadotropin secretion could be of great interest for the development of an improved contraceptive.


Inhibition of growth of a murine sarcoma by immunization with chemically modified cells. Sarcoma I cells from ascitic fluid of A/Ph mice were modified with iodoacetamide, diketene and glutaraldehyde. Syngeneic and allogeneic mice were immunized with the modified cells and the effect of this immunization on the growth or rejection of the native sarcoma injected intramuscularly into the hind leg was ascertained. The antitumour effect of the modified cells was compared with that of the irradiated ones. Only cells treated with iodoacetamide gave a better antitumour effect than the unmodified cells irradiated with a minimal dose of gamma irradiation capable of stopping the proliferation of these cells.


[Effect of mezaton on experimental myocardial infarct with arrhythmias in rabbits]. In 35 of 40 rabbits operated upon experimental myocardial infarction (EMI) was reproduced. Immediately after legation of coronary arteries (CA) and during the next 2 days mesaton was introduced in a single dose into the marginal auricular vein of 12 rabbits with EMI. The results of the antiarrhythmic effect produced by the drug were judged on the basis of the dynamic ECG findings in three standard leads. From the materials thus obtained it followed that a single administration of mesaton during an acute period of EMI (the first 3 days of the experiment) prevented the development of composite disturbances of the rhythm and reduced lethality among the animals. A repeated introduction of mesaton to rabbits with EMI, but without any arrhythmia provoked ventricular extrasystole in 2 and sinus bradycardia in one of the 12 animals. Repeated  administration of mesaton to rabbits with EMI complicated by ventricular extrasystole did not eliminate the upset function of excitation, while with the ventricular extrasystole transforming into ventricular tachycardia the eventual prognosis becomes unfavourable.


[Several criteria of evaluation of foot-and-mouth disease virus reproduced in cell cultures in suspension]. The study of the plaques produced by viruses Asia and O1 reveals different properties which may depend on the virus or on the cell or its method of culture. Antigenic differences and subsequent immunological differences correspond to differences in plaques. On the other hand, the cell line in suspension evolves both in its morphology and its receptivity during the passages. All these variations imply the observations of quantitative and qualitative criteria of evaluation in order to prepare vaccines from viruses similar to viruses specific to a given epizooty.


[Studies on the diffusion of cephapirin into prostatic tissue]. After i.v.-application of 2 grams of Cephaprin (= Bristocef) to 13 patients with prostatic adenoma the mean serum concentrations after 30 minutes were 50 microgram/ml and 21.5 microngram/ml after 60 minutes. The corresponding tissue values were 20.8 and microgram/g respectively in prostatic tissue. The evaluated serumand tissue cncentrations are suited to the therapy of infections with cephapirin sensitive rods.


Inheritance of Ag-stainability of nucleolus organizer regions. Investigations in 7 families with trisomy 21. The Ag-stainability of the nucleolus organizer region (NOR) was studied in the acrocentric chromosomes identified by Q-banding in cultured lymphocytes from seven children with trisomy 21 and their parents. The observed Ag-NOR patterns were in accordance with chromosomal inheritance except for a slight intraindividual variation which might be explained mainly by technical causes. In two cases the meiotic nondisjunction could be attributed to one of the parents, once to the father, and once to the mother. It is concluded that the Ag-stainability of the NORs is in general a heritable characteristic of the acrocentric chromosomes in maximally activated cells as, e.g., cultured lymphocytes. It may reflect individual differences in the amount of rDNA as well as differences in the capacity for NOR activation.


The world distribution of the electrophoretic variants of the red cell enzyme esterase D. The phenotypic variation in the esterase D phenotypes among 2,405 individuals in 14 samples from populations in Europe, Africa and Asia are reported. There exists a marked difference in esterase D allele frequencies in different continental regions. Comparison of the world population data so far available show that esterase D is another useful genetic parameter for the study of population diversity.


Restriction of the anti-bovine serum albumin response in rabbits immunized with Micrococcus lysodeikticus. Rabbits capable of producing antibodies of restricted heterogeneity in response to Micrococcus lysodeikticus are equally capable of producing antibodies of restricted heterogeneity to bovine serum albumin. These antibodies are produced when animals are simultaneously injected with micrococcus and BSA and their specificity is restricted to a small number of epitopes. These results suggest that micrococcal vaccines can induce the restriction of heterogeneity in antibodies raised against totally unrelated antigens.


Expression of hepatitis B virus-specific markers in asymptomatic hepatitis B surface antigen carriers. A study was undertaken to assess the state of hepatitis B virus infection in a group of asymptomatic hepatitis B surface antigen (HBsAg) carriers. This study confirmed that the presence of hepatitis B e antigen (HBeAg) in serum was closely associated with serum HBsAg-specific deoxyribonucleic acid polymerase activity, hepatitis B core antigen (HBcAg) in serum and liver cell nuclei, and a histological picture of chronic hepatitis. No HBsAg-specific deoxyribonucleic acid polymerase activity or HBcAg was detected in highly concentrated anti-HBe-positive sera. In addition, liver biopsy specimens from carriers with anti-HBe were negative for HbcAg by immunofluorescence, and the liver histology was either normal or revealed only fatty changes. These data indicate that the anti-HBe-positive sera contained either no Dane particles or, if present, at least a 500-fold-lower concentration of Dane particles than that found in HBeAg-positive sera.


Radioimmunoassay for the antigenic determinants of cholera toxin and its components. A radioimmunoassay procedure is described for the detection of cholera toxin and its component polypeptide chains. Cholera toxin, A subunit, B subunit, alpha chain, and gamma chain were iodinated by the chloramine T procedure. Radiolabeling did not significantly alter the polyacrylamide electrophoretic migration patterns of the toxin or its components. Moreover, radiolabeled toxin, B subunit, and alpha chain preparations retained substantial ability to bind to intestinal mucosal homogenates. The minimal amount of antitoxin detectable with radiolabeled toxin was 0.04 antitoxin units/ml. Substitution of radiolabeled B subunit, A subunit, and alpha chain for radiolabeled toxin decreased the sensitivity of the test. Radiolabeled gamma chain did not bind to the antitoxin preparation. Competitive inhibition studies, with titrated anti-choleragen serum and radiolabeled toxin or components, indicated that the minimum concentration of toxin detectable was 7.0 x 10(-8) mumol/ml at a 90% inhibition level. The A subunit and alpha chain preparations inhibited the binding of the radiolabeled B subunit to antitoxin sites. Conversely, B subunit inhibited the binding of radiolabeled A subunit and alpha chain to antitoxin. The gamma chain did not show any reaction with antitoxin or cross-reaction with either whole toxin or its components. These results strongly suggest that the A subunit and the alpha chain contain antigenic determinant(s) that are common to the B subunit. The B subunit (beta chain) and the alpha chain of cholera toxin may therefore contain region(s) of chemical similarity.


Effect of heat on antigenicity and immunogenicity of the antigenic determinant shared by Haemophilus influenzae type b and Escherichia coli K100. Escherichia coli K100 produces an antigenic determinant similar to, or identical with, the capsular antigen of Haemophilus influenzae type b. Studies of the effects of heat on the immunogenicity, erythrocyte-modifying capacity, and antigenicity of this cross-reacting antigen (CRA) revealed the following findings. Immunization of rabbits with viable or formaldehyde-killed suspensions of E. coli K100, producing CRA, engendered CRA antibodies in significant titers, as demonstrated by hemagglutination of erythrocytes modified by H. influenzae type b antigen. Heating of the suspensions for 1 h at 56 or 100 degrees C destroyed the immunogenicity of CRA, and the heated suspensions did not prime for a secondary antibody response. Supernatants of heated suspensions also were non-immunogenic. Repeated freezing and thawing of heated suspensions of E. coli K100 or their supernatants did not restore immunogenicity. Heat also abolished the immunogenicity of H. influenzae type b. The loss of immunogenicity of CRA of E. coli K100 by heat was not due to alteration of the antigenic determinant, since heated suspensions and supernatants thereof modified erythrocytes for agglutination by H. influenzae type b antiserum. The latter supernatants also inhibited hemagglutination by H. influenzae type b antibodies and absorbed the latter. We conclude that striking differences exist in the effects of heat on CRA on the one hand and of enterobacterial common antigen and lipopolysaccharide O antigen of enteric bacteria on the other. Heating of the latter two antigens does not abolish their priming effect, and repeated freezing and thawing restores the immunogenicity of heated antigens.


RNA-reverse transcriptase complex from cultured human myeloma-leukemia cells. A high molecular weight RNA-reverse transcriptase complex in the culture media of peripheral leukocytes obtained from two Japanese patients with myeloma-leukemia was detected by demonstration of a 3H-uridine peak and a peak of DNA polymerizing activity banding at a density of 1.15-1.19g/ml. The enzyme in the complex was able to utilize poly(rA)-d(pT)10 or poly (rC)-d(pG) 12-18, but not poly (dA)-d(pT) 10 or (dT) 12-18 as template-primers. The sucrose density sedimentation analysis revealed that RNA in the complex sedimented at a location of approximately 50s and 20-30s.


Nucleic acid content and cell cycle distribution of five human bladder cell lines analysed by flow cytofluorometry. Five human bladder cell lines, four derived from tumor tissue and the fifth originating from presumed benign transitional epithelium, were examined by flow cytofluorometry to estimate the DNA and RNA content per cell during exponential and stationary phases of growth. A new staining technique was employed using acridine orange to differentially stain DNA and RNA in unfixed cells made permeable to the dye and other reagents by treatment with detergent at low ph. Stemline chromosome numbers for each cell line correlated well with relative DNA content of the G1 population as measured by this technique. In addition, the simultaneous measurements of DNA and RNA per cell yielded cell cycle distributions for each cell line. The ratio of stainable RNA/DNA was lower for all cell lines derived from bladder tumors as compared to the presumed normal cell line, indicating high nuclear/cytoplasmic ratios for the former.


Antigenic differences between primary methylcholanthrene-induced rat sarcomas and post-surgical recurrences. The immunogenicities of in vivo lines established from primary MCA-induced rat sarcomas have been compared with those of lines initiated from tumour recurrences at the site of the primaries' surgical excisions. Lines from two of four primary sarcomas showed little or no immunogenicity, as assessed by protection against challenge afforded by graft excision or implantation of irradiated tissue. In contrast, lines from all four recurrences were immunogenic, giving protection against up to 5 X 10(6) tumour cells. Most importantly, with all four tumours, lines established from recurrences were antigenically distinct from lines derived from their original primary sarcomas, so that immunization with regrowth lines gave no protection against the lines from the primaries, and vice versa. These studies demonstrate that primary MCA-induced sarcomas are antigenically distinct from recurrent tumors arising after surgical removal of the primaries, implying that these tumours arose by clonal amplification of separate populations of transformed cells. This may reflect proliferation of dormant neoplastic cells or the further induction of transformed cells by residual carcinogen. These findings are relevant to the multifocal origin of tumours and for the design of active immunotherapy for the treatment of recurrences or metastatic deposits.


SV40 tumor rejection induced by vesicular stomatitis virus bearing SV40 tumor-specific transplantation antigen (SV40-TSTA). I. Specificity of immunoprotection and effect of enzyme treatment on TSTA activity. Highly purified vesicular stomatitis virus (VSV) was obtained from VSV-infected SV40-transformed hamster cell lines. Immunization with this virus protected hamsters against challenge with SV40-transformed cells (TSV5-cl2). This protection was obtained regardless of the source of the SV40-transformed cells (e.g. cat, rat, hamster) used to produce VSV, and was therefore associated with the SV40 tumor-specific transplantation antigen (SV40-TSTA). Furthermore, when grown on spontaneously transformed cell lines or on cells transformed by a different oncogenic DNA virus, such as polyoma virus, the VSV failed to protect against the SV40-induced tumor. It was concluded that the SV40-TSTA activity of purified VSV is due to the incorporation of SV40-TSTA within the viral envelope. When VSV was treated with proteolytic enzymes (bromelain, trypsin) no loss of TSTA-induced tumor rejection was observed, although VSV had lost its ability to induce virus-neutralizing antibody. This clearly demonstrates that the TSTA activity is not related to the viral spikes. Phospholipase C suppressed the TSTA activity but neutralizing activity was still detectable in the anti-VSV sera. The results presented here demonstrate that the protection afforded by VSV is highly specific. It is particularly interesting that SV40-TSTA activity may be conveyed by the lipid core of the viral envelope.


SV40 tumor rejection induced by vesicular stomatitis virus bearing SV40 tumor-specific transplantation antigen (SV40-TSTA). II. Association of SV40-TSTA activity with liposomes containing VSV glycolipids. Highly purified vesicular stomatitis virus (VSV) was obtained from VSV-infected SV40-transformed and from "normal" hamster cell lines. A glycolipid extract was prepared from these VSV preparations according to the Folch partition procedure. These glycolipids were rendered immunogenic to the Syrian hamsters when incorporated within liposomal membranes composed of lecithin/sphingomyelin/cholesterol (1/1/2 by weight). When the glycolipids were extracted from VSV grown on cell lines (TSV5-cl2 and EHSVi-cl1) which contained the SV40 tumor-specific transplantation antigen (SV40-TSTA), it was possible either to induce a tumor rejection or at least to slow the growth of the tumor in Syrian hamsters challenged with TSV5-cl2 cells. No protection was obtained in animals treated with liposomes containing glycolipids extracted from purified VSV grown on SV40-TSTA-negative cells (EHB). The SV40-TSTA could be a glycolipid of the transformed cell membrane which is incorporated within the VSV envelope.


Separation of the Moloney leukemia virus-determined cell surface antigen (MCSA) from known virion proteins associated with the cell membrane. Cell membranes of Moloney lymphoma cells (YAC, of strain A origin) were solubilized by NP40. The antigenicity of the solubilized protein fraction was assayed by inhibition of the corresponding cytotoxic reaction against YAC target cells. The Moloney leukemia virus (MLV)-determined cell surface antigen (MCSA) was detected with mouse antisera, produced by the repeated inoculation of heavily irradiated YAC cells into syngeneic mice. Virion proteins gp71, p30, p15, p12 and p10 were identified with goat or rabbit antisera against purified Rauscher and Friend leukemia virus proteins. MCSA was found to bind to Con-A--Sepharose and was eluted by mannoside together with H-2A AND GP71. In contrast, p30, p12, p10 and part of p15 and p15(E), were not retained on the column and could be separated from MCSA. Passage of the glycoprotein fraction through Sephadex G-200 led to the separation of MCSA activity from gp71 and H-2A. MCSA eluted between the immunoglobulin (IgG) and the bovine serum albumin (BSA) size markers. MCSA could be also separated from the known viral proteins and from H-2 by velocity centrifugation in sucrose gradients. It sedimented with approximately 6.6 S ahead of gp71 (4.4 S) and H-2 (3.2 S). It is suggested that MCSA may be a glycoprotein with an approximate molecular weight of 110,000 and distinct from the known viral proteins gp71, p30, p15(E), p12, p10 and from H-2.


Experimental yolk-sac tumors produced by fetectomy without virus infection in rats. Pregnant WKA rats were laparotomized on the 12th day of gestation, and all fetuses were removed from the uteri, leaving the fetal membranes attached to placentas in situ, protruding into the peritoneal cavity. After 5 to 6 months, extrauterine tumors were observed in 80% of the operated rats, and elevated serum alpha-fetoprotein levels were detected in 80% of the tumor-bearing rats. The tumors in AFP-positive rats always contained yolk-sac tumor elements. In transplantable tumor lines established in the ascites form, numerous free-floating embryoid bodies were observed. The experiments demonstrate that yolk-sac tumors (endodermal sinus tumors) can be induced by simple fetectomy without infection by exogenous tumor virus.


Detection of herpes simplex virus-related antigens in the nuclei and cytoplasm of biochemically transformed cells with peroxidase/anti-peroxidase immunological staining and indirect immunofluorescence. Herpes simplex virus (HSV)-related antigens have been demonstrated in the nuclei and cytoplasm of human and mouse cells biochemically transformed by ultraviolet light-irradiated HSV. This was accomplished by using peroxidase/anti-peroxidase immunological staining and indirect immunofluorescence with rabbit antisera that had high neutralizing titers against the HSV-specific thymidine kinase activity and virus infectivity. HSV-1 antisera reacted with antigens in cells biochemically transformed by type 1 HSV, but not with those of cells biochemically transformed by type 2 HSV. Similarly, HSV-2 antisera reacted with antigens in cells biochemically transformed by HSV-2, but not with those in cells biochemically transformed by HSV-1. In contrast, herpes virus-related antigens were detected in cells cytolytically infected with HSV-1 and with HSV-2 by either type 1 or type 2 HSV antisera. These observations suggest that the antigens detected in the biochemically transformed cells were a type-specific subset of the HSV-related antigens synthesized in cells undergoing productive infection by HSV-1 and HSV-2.


alpha-fetoprotein-containing cells in the early stages of liver carcinogenesis induced by 3'-methyl-4-dimethyl-aminoazobenzene and 2-acetylaminofluorene. The morphology of rat liver cells containing alpha-feto-protein (AFP) during early stages of carcinogenesis induced by 3'methyl-4-dimethylaminoazobenzene and 2-acetylaminofluorene was investigated. Indirect immunofluorescence was used to detect AFP. AFP was not found in the cells of hyperplastic nodules but was present in the cells located in areas of transitional cell proliferation. A large proportion of the AFP-positive cells formed gland-like structures. The cells containing AFP were at various levels of differentiation according to morphological criteria. Poorly differentiated, small, basophilic cells were predominant amont the AFP-positive population. The most highly differentiated AFP-positive cells had the morphology of hepatocytes.


The histochemical demonstration of human prostatic acid phosphatase with phosphorylcholine. Tissues from patients with benign prostatic hyperplasia were used for the cytochemical demonstration in light and electron microscopy of a secreted, nonlysosomal prostatic acid phosphatase (PAP) with phosphorycholine, substrate specific for PAP. The specificity of phosphorylcholine for PAP is attributable to the pentavalent nitrogen in phosphorylcholine, a feature that renders it resistant to hydrolysis by all other acid phosphatases. PAP activity was found in the Golgi cisternae and its associated vacuoles and in secretory vacuoles localized in the nuclear, Golgi, and apical areas of the prostatic epithelial cell. These results confirm the existence of two types of acid phosphatase in prostatic tissue. One is lysosomal and is prevalent in many tissues and the other, PAP, is the major enzymatic product secreted by the prostate. The specificity of PAP for phosphorylcholine, one of the natural substrates for this enzyme, validates the use of this method for the histochemical characterization of PAP and indicates the prostatic origin of cells showing PAP activity.


The utility of antisera to canine growth hormone and canine prolactin for immunocytochemical staining of the dog pituitary gland. Using the immunoperoxidase technique and specific homologous antisera to highly purified canine prolactin (PRL) and canine growth hormone (GH), selective immunocytochemical staining was localized in two different cell types in the pars distalis of the dog pituitary gland. The specificity and validity of both antisera were demonstrated by elimination of their immunostaining capacify after prior absorption with their respective antigens, while absorption with other antigens failed to decrease staining intensity. This was further confirmed by studying the immunologic behavior of both antisera in the agar gel double diffusion immunoprecipitin test and by competitive radioactive binding analysis. The specific heterologous antisera to ovine PRL and human GH were shown to be just as useful as antisera to canine hormones for immunocytochemical demonstration of the individuality of PRL and GH cells in the dog pituitary. This confirms the validity and specific of our hitherto heterologous immunocytochemical procedure. However, because of their high potency the anti-canine GH and anti-canine PRL sera seem to be more efficient for immunocytochemical staining of the dog pituitary. The immunoreactive PRL and GH cells were also distinguished with reasonable certainity in two different populations of acidophils according to their tinctorial affinity. Both cell types differed further from each other in respect of their relative proportion, morphology and topography, which were variously pronounced for each sex.


Histochemistry of carboxyl esterases in the broad bean root tip with indoxyl substrates. The possibility of the use of indoxyl substrates for enzyme localization in plant material was tested and proved. Studying the localization of the carboxyl esterase in broad ben root tip a) the time course of the indigogenic reaction was investigated, b) the concentration of the ferro/ferri redox buffer was examined, c) various substances were used to control the indigogenic visualizing reaction, d) the azocoupling procedure was applied with indoxyl substrates, e) unhalogenated and halogenated substrates were tried, and, finally, f) acetate and butyrate were employed in the indigogenic procedure. Using the last mentioned (f) trial the existence and localization was verified in the material tested of carboxyl esterase fractions recognized there previously on the basis of azocoupling reactions with naphtholic substrates.


Osmium tetroxide/p-phenylenediamine staining of nucleoli and Balbiani Rings in Chironomus salivary glands. This paper deals with the application of the osmium tetroxide fixation followed by p-phenylenediamine treatment to salivary gland cells from Chironomus larvae. After this procedure, cytoplasm, nucleoli and Balbiani rings show a high degree of staining both in light and electron microscopy, while chromatin remains unstained. Ethanol fixation followed by osmium tetroxide/p-phenylenediamine does not modify the above mentioned staining pattern. Under these conditions, extractive procedures for lipids do not affect the osmiophilia of nucleoli and Balbiani rings, while RNase or trichloroacetic acid treatment decreaes the staining degree of these structures. In osmium tetroxide/p-phenylenediamine treated salivary glands, the highest contrast within nuclei is seen to occur in the pars granulosa from normal or segregated nucleoli, as well as in Balbiani ring granules, which appear either as hollow granules or with a bipartite or horseshoe-like structure.


Peroxidase and pseudoperoxidase reactions in relation to sudanophilia. A selective staining of hemoglobin in erythroid cell series was achieved by use of Sudan Black B (modified method of Sheehan and Storey) if optimal amount of hydrogen peroxide was added to the staining mixture. The effect of some inhibitory agents (KCN, wet heat, pH) on this staining as well as on the Lepehne's pseudoperoxidase reaction for hemoglobin was similar. Both reactions were more resistant to these factors than the peroxidase reactions and sudanophilia in granulocytes in which both could be blocked by the pretreatment with absolute methanol. Moreover the effect of some extraction procedures for lipids on both myeloperoxidase reactions and sudanophilia was investigated. The results support the view that the sudanophilia in granulocytes is due to their peroxidase activity and for the staining of hemoglobin by use of Sudan Black B with H2O2 its pseudoperoxidase activity is responsible. In addition the effect of the substitution of phenolphosphate by dihydroxybenzenes on granulocyte sudanophilia is reported.


Does diaminobenzidine demonstrate prostaglandin synthetase? A study on polyunsaturated fatty acid-induced DAB oxidation in sheep vesicular glands and rabbit kidney medulla. A method histochemical localization of prostaglandin synthetase using DAB, potassium cyanide and polyunsaturated fatty acid has been revised. The arachidonic acid-induced DAB oxidation observed in the secretory epithelium of sheep vesicular glands and in collecting tubules as well as intersititial cells of rabbit kidney medulla was found to be insensitive to antiinflammatory cyclooxygenase (formerly referred as prostaglandin synthetase) inhibitors, such as indomethacin, aspirin, mefenamic acid and paracetamol, whereas aminotriazole caused complete inhibition of the reaction. Furthermore, DAB was oxidized in the presence of polyunsaturated fatty acids inconvertible to prostaglandins (linoleic and linolenic acid) as well as in the presence of H2O2--in the latter case reaction possessed identical features with that induced by fatty acids. Ultrastructurally, the reaction product was localized on the membranes of nuclear envelope and endoplasmic reticulum. On the ground of the results obtained a hypothesis is presented, that the polyunsaturated fatty acid-induced DAB oxidation is due to a peroxidatic activity of the investigated tissues. Possible relations between such peroxidatic activity and prostaglandin biosynthesis are discussed.


The early development of autistic children. A sample of 74 young autistic children was selected and defined by direct observation of specific behaviors and clinical assessment of the presence or absence of associated pathological conditions. Retrospective developmental data on these children and 38 age-matched normal children were gathered by means of a written inventory completed by the parents when the children were relatively young (mean age less than 4 years). The autistic children were reported to have had significant delays in the development of motor abilities, speech, communication, comprehension, and, to a lesser extent, perception during their 1st and 2nd years.


List of descriptive cataloging elements for use in bibliographic control of nonprint materials. Bibliographic control of audiovisuals or nonprint materials, generated at the point of publication, is almost nonexistent. The authors address this problem and present an outline of bibliographic elements that nonprint producers need to include on each individual production.


Legal and ethical aspects of proprietary rights to federally funded media programs. This article attempts to place in proper perspective the legal and ethical aspects of proprietary rights as they apply to federally funded media programs. Brief reference is made to the recently enacted revision of the Copyright Law, Title 17 of the U.S. Code. Distinction is made between the common law and statutary copyright protection afforded to the creator of an original work. The 'Fair Use' doctrine and the 'Rights in Data' clauses of the general provisions of the Federal Procurement Regulations are discussed in detail; so is the use of Talent Consent and Waivers and Easements of Copyright.


Organization and cellular biology of the perichondrial ossification groove of ranvier: a morphological study in rabbits. The perichondrial ossification groove of Ranvier, a circumferential groove in the periphery of the epiphyseal cartilage, was studied in rabbits whose ages ranged from one week to eight months using light and electron microscopy, autoradiography after labeling with 3H-thymidine, 3H-proline, and 3H-glucosamine, and histochemical staining for proteoglycans and alkaline phosphatase. By these methods, three groups of cells were identified within the groove: 1. A group of densely packed cells deep in the groove, which are the progenitor cells for the osteoblasts that form the bone bark, a cuff of bone surrounding the epiphyseal growth-plate region and the adjacent part of the metaphysis. 2. A group of more widely dispersed, relatively undifferentiated mesenchymal cells and fibroblasts, some of which are chondroblast precursors that probably contribute to appositional chondrogenesis and growth in width of the epiphyseal cartilage. 3. Fibroblasts and fibrocytes among sheets of highly oriented and organized collagen fibers which form a fibrous layer that is continuous with the outer fibrous layer of the periosteum and with the perichondrium. This layer also sends fibers into the epiphyseal cartilage and anchors the periosteum firmly to the epiphyses as bone growth proceeds.


Plasma membrane folds on the mast cell surface and their relationship to secretory activity. Changes in the surface morphology of secreting mast cells have been followed by scanning electron microscopy. Mast cells isolated from the rat peritoneal cavity have folds of plasma membrane that form snake-like ridges on their surfaces. Fold length varies considerably from cell to cell, whereas fold width and depth appear to remain relatively constant. To assess the possible relationship between secretory activity and surface folding, a seimquantitative method was used for measuring fold length in control and secreting populations. A positive correlation is found between secretion of histamine and the extent of membrane folds on the mast cell surface. The source of the membrane required for fold formation is probably secretory granule membrane incorporated into the plasma membranene as a result of exocytosis. Furthermore, a distinct cell type devoid of surface folds, designated as a raspberry-type cell, is found to occur as an integral part of a normal population of mast cells. This cell type is resistant to stimulation by polymyxin.


The preservation of ultrastructure in saturated phosphatidyl cholines by tannic acid in model systems and type II pneumocytes. The preservation for electron microscopy of saturated phospholipids in general, and phosphatidyl choline (PC)in particular, remains and unsolved problem since OsO(4) and glutaraldehyde are incapable of interacting with PC directly. However, by introducing tannic acid preceding osmication, we were able to demonstrate highly ordered, preserved lamellar structures in model experiments with saturated PC, and in vivo experiments type II pneumocytes of lung tissue. The secretory bodies of the latter are known to contain a high proportion of these saturated phospholipids. In both cases, the repeating periodicity approximated 45 A. It was determined that tannic acid interacts with the choline component of PC to form a "complex," which then could be stabilized by treatment with OsO(4). In the absence of osmication, the PC-tannic acid complex acid did not survive conventional dehydration techniques, but osmication permitted conventional Epon embedment. Sphingomyelin (SPH), which contains choline, behaved similarly in model experiments. But there was no evidence of a comparable reaction with tannic acid using phosphatidyl ethanolamine (PEA), phosphatidyl serine (PS), or phosphstidy inositol (PI). Chemical studies indicted a high pH dependency for the formation of the PC- tannic acid complex. Also, experiments demonstrated its dissociation in various organic solvents. Sharp delineation and great contrast of the polar zones in the ordered lamellar structures was achieved by additional staining with lead citrate thus leading to the conclusion that tannic acid serves as a multivalent agent, capable of simultaneous interaction with saturated PC, OsO(4), and lead citrate stains.


Polymorphism of myosin among skeletal muscle fiber types. An immunocytochemical approach was used to localize myosin with respect to individual fibers in rat skeletal muscle. Transverse cryostat sections of rat diaphragm, a fast-twitch muscle, were exposed to fluorescein-labeled immunoglobulin against purified chicken pectoralis myosin. Fluorescence microscopy revealed a differential response among fiber types, identified on the basis of mitochondrial content. All white and intermediate fiber but only about half of the red fiber reacted with his antimyosin. In addition, an alkali-stable ATPase had the same pattern of distribution among fibers, which is consistent with the existence of two categories of red fibers. The positive response of certain red fibers indicates either that their myosin has antigenic determinants in common with "white" myosin, or that the immunogen contained a "red" myosin. Myosin, extracted from a small region of the pectorlis which consists entirely of white fibers, was used to prepare an immunoadsorbent column to isolate antibodies specific for white myosin. This purified anti-white myosin reacted with the same fibers of the rat diaphragm that had reacted with the white, intermediate, and some red fibers are sufficiently homologous to share antigenic determinants. In a slow-twitch muscle, the soleus, only a minority of the fiber reacted with antipectoralis myosin. The majority failed to respond; hence, they are not equivalent to intermediate fibers of the diaphragm; despite their intermediate mitochondrial content. Immunocytochemical analysis of two different musles of the rat has demonstrated that more than one isoenzyme of myosin can exist in a single muscle, and that individual fiber types can be recognized by immunological differences in their myosin. We conclude that, in the rat diaphragm, there are at least two immunochemically distinct types of myosin and four types of muscle fibers: white, intermediate, and two red. We suggest that these fibers correspond to the four types of motor units described by Burke et al. (Burke, R. E., D. N. Levine, P. Tsairis, and F. E. Zajac, III 1973. J. Physiol. (Lond) 234:723-748.)in the cat gastrocnemius.;


Myosin subfragment binding for the localization of actin-like microfilaments in cultured cells. A light and electron microscope study. Fluorescein-labeled heavy meromyosin subfragment-1 (F-S-1) has been purified by ion exchange chromatography and characterized in terms of its ability to bind specifically to actin. F-S-1 activates the Mg++-adenosine triphosphatase activity of rabbit skeletal muscle actin and decorates actin as shown by negative stains and thin sections of rabbit actin and rat embryo cell microfilament bundles, respectively. Binding of F-S-1 to cellular structures is prevented by pyrophosphate and by competition with excess unlabeled S-1. The F-S-1 is used in light microscope studies to determine the distribution of actin-containing structures in wnterphase and mitotic rat embryo and rat kangaroo cells. Interphase cells display the familiar pattern of fluorescent stress fibers. Chromosome-to-pole fibers are fluorescent in mitotic cells. The glycerol extraction procedures employed provide an opportunity to examine cells prepared in an identical manner by light and electron microscopy. The latter technique reveals that actin-like microfilaments are identifiable in spindles of glycerinated cells before and after addition of S-1 or HMM. In some cases, microfilaments appear to be closely associated with spindle microtubles. Comparison of the light and electron microscope results aids in the evaluation of the fluorescent myosin fragment technique and provides further evidence for possible structural and functional roles of actin in the mitotic apparatus.


Localization of thyroiglobulin antigenicity in rat thyroid sections using antibodies labeled with peroxidase or 125I-radioiodine. In the hope of localizing thyroglobulin within focullar cells of the thyroid gland, antibodies raised against rat thyroglobulin were labeled with the enzyme horseradish peroxidase or with (125)I-radioiodine. Sections of rat thyroids fixed in glutaraldehyde and embedded in glycol methacrylate or Araldite were placed in contact with the labeled antibodies. The sites of antibody binding were detected by diaminobenzidine staining in the case of peroxidase labeling, and radioautography in the case of 125(I) labeling. Peroxidase labeling revealed that the antibodies were bound by the luminal colloid of the thyroid follicles and, within focullar cells, by colloid droplets, condensing vacuoles, and apical vesicles. (125)I labeling confirmed these findings, and revealed some binding of antibodies within Golgi saccules and rough endoplasmic reticulum. This method provides a visually less distinct distribution than peroxidase labeling, but it allowed ready quantitation of the reactions by counts of silver grains in the radioautographs. The counts revealed that the concentration of label was similar in the luminal colloid of different follicles, but that it varied within the compartments of follicular cells. A moderate concentration was detected in rough endoplasmic reticulum and Golgi saccules, whereas a high concentration was found in condensing vacuoles, apical vesicles, and in the luminal colloid. Varying amounts of label were observed over the different types of colloid droplets, and this was attributed to various degrees of lysosomal degradation of thyroglobulin. It is concluded that the concentration of thyroglobulin antigenicity increases during transport from the ribosomal site of synthesis to the follicular colloid, and then decreases during the digestion of colloid droplets which leads to the release of the thyoid hormone.


New human adenovirus (candidate adenovirus type 35) causing fatal disseminated infection in a renal transplant recipient. An antigenically distinct adenovirus is described which was isolated in March 1973 from the lungs and kidney of a 61-year-old woman who died of diffuse interstitial adenovirus pneumonia 55 days after receiving a cadaveric renal allograft. Complement fixation, hemagglutination inhibition, and serum neutralization tests on sequential serum specimens from the patient confirmed that the adenovirus infection occurred in coincidence with her clinical illness and failed to document concomitant infection by any other common respiratory agent. Pathological and virological findings indicated that the pneumonia was only one manifestation of a disseminated adenovirus infection, the source of which may  have been a latent infection pre-existing in the donor kidney. The adenovirus, purified by terminal dilution and plaque procedures, has antigenic, morphological, biological, biophysical, host susceptibility, and hemagglutinating properties characteristic of adenovirus group 1A. Buoyant densities in  CsCl are 1.340 g/ml for the virion, 1,300 g/ml for the group complement-fixing (hexon) antigen, and 1.290 g/ml for the  major soluble complete hemagglutinin (dodecon). The virus was serologically distinct from adenoviruses 1 to 34 in reciprocal serum neutralization tests with antisera to these viruses. We propose this virus as candidate adenovirus type 35 (holden).


Early detection of chlamydial inclusions combining the use of cycloheximide-treated McCoy cells and immunofluorescence staining. Detection of Chlamydia trachomatis inclusions only 21 h after a specimen reaches the laboratory has been achieved by the combined use of cycloheximide-treated McCoy cells and immunofluorescence staining. Moreover, cells exposed to cycloheximide were more sensitive for detecting chlamydial inclusions than those pretreated by irradiation, since larger numbers of inclusions were found in the former cells. The application of this rapid and sensitive method allows a diagnosis of chlamydial infection to be made before antibiotic therapy is started. In this way, it should enable the treatment of nonspecific genital infections to be placed on a more rational basis.


Synthesis and secretion of alpha-2-macroglobulin by cultured adherent lung cells. Comparison with cell strains derived from other tissues. We studied the synthesis and secretion of alpha-2-macroglobulin by cultures of human adherent cells. Much more alpha-2-macroglobulin (measured by radioimmunoassay) accumulated in media of established strains of adherent cells derived from embryonic lung than in media of established strains derived from adult skin or rheumatoid synovium. Alpha-2-macroglobulin accumulated in media of primary cultures of adherent cells from a variety of embryonic tissues. However, the amount of alpha-2-macroglobulin accumulating in media of subsequent passage of these cells declined for all strains except those derived from lung. Immunodiffusion and double-antibody immunoprecipitation studies of cell extracts and media after incubation of cells with l-[(35)S]methionine supported the radioimmunoassay finding that adherent cells from lung synthesized and secreted more alpha-2-macroglobulin than adherent cells from skin. Intracellular alpha-2-macroglobulin could not be detected by radio-immunoassay or visualized by immunofluorescent microscopy, suggesting that synthesized alpha-2-macroglobulin is rapidly secreted. Plasminogen-rich fibrin clots were lysed in culture media of adherent cells from embryonic lung and, to a lesser extent, heart. Adherent cells from other tissues, which produced less alpha-2-macroglobulin, did not lyse fibrin clots. However, all cultures of adherent cells contained pericellular fibronectin, a large, external, transformation-sensitive glycoprotein known to be cleaved by plasmin. We speculate that production of alpha-2-macroglobulin may be a means for protease-secreting normal cells to preserve cell surface integrity and that alpha-2-macroglobulin synthesized locally in lung may protect lung tissues from a variety of proteases.


Separation of fucosyltransferase and endogenous acceptors of fucose from human parotid saliva. Using gel filtration on Sephadex G-100 and column chromatography on CM-cellulose, fractions containing either endogenous acceptor of L-fucose or fucosyltransferase were obtained from human parotid saliva. Hydrolysis with mild acid followed by Smith degradation increased the fucosyltransferase activity of the fractions that contained endogeneous acceptor.


Polistes wasp hypersensitivity: diagnosis by venom-induced release of histamine in vitro. Polistes wasps cause a majority of Hymenoptera-induced anaphylactic reactions in Texas. Using the in vitro release of histamine from basophils of patients allergic to Polistes stings, we have studied the cross-reactivity of venoms from three species of Polistes wasps as well as the cross-reactivity among Polistes, honeybee, and Vespula maculifrons (yellow jacket) venoms. Venom collected by an extrusion technique from Pollistes exclamans, Pollistes apachus, and Pollistes carolina caused release of histamine in seven Polistes-sensitive individuals. The dose-response curves from all three Polistes species were quite similar, suggesting extensive cross-reactivity among these species. None of these patients showed significant release of histamine from leukocytes exposed to yellow jacket or honeybee venom. We conclude that a source of Polistes venom is available for further study and possibly for therapy. It appears that any of three local common species of Polistes wasps could be used. Our studies confirmed earlier reports that Hymenoptera sensitivity if often genus-specific.


On the lack of specificity of the cobalt-bicarbonate method for carbonic anhydrase. Data are presented that support a nonenzymic mechanism for the staining obtained with the cobalt-bicarbonate method. The biochemically inactive apocarbonic anhydrase and Cu+2 apocarbonic anhydrase stain positively and this stain is inhibited by acetazolamide. The staining of the acetazolamide resistant carbonic anhydrase of male rat liver is inhibited by 10-6 M acetazolamide, at which concentration no biochemical inhibition is observed. There is no correlation between the biochemical and histochemical inhibitory potencies of a number of sulfonamides. The nonsulfonamide inhibitor, KCNO, does not inhibit staining. When incubations are performed in media exposed to atmospheres of increasing CO2 content, staining is not abolished until the atmospheric pCO2 approaches that generated by the medium itself. This finding renders the carbonic anhydrase catalyzed dehydration of HCO3- an improbable reaction for the staining. Studies with modified media show differences in staining patterns and in sensitivity to acetazolamide inhibition which question the specificity of the method for carbonic anhydrase.


A moving spot high resolution scanning microspectrometer using focused laser illumination: illustrations with acriflavin stained sephadex beads and human fibroblast nuclei. We describe a high resolution moving spot scanning microspectrometer, capable of absorption or fluorescence detection, using focused laser illumination which is moved over the sample by rotating the laser beam direction prior to focusing. This rotation is achieved by reflecting the beam from mirrors mounted on bending mode piezoelectric transducers which, when bent by an applied voltage, cause the mirrors to rotate. The images of optically thin samples are analyzed by considering the convolution of the focused spot intensity distribution with the absorbance of a uniformly stained spherical particle. This analysis is verified experimentally with data from acriflavin stained Sephadex beads. Data from acriflavin-Feulgen stained human fibroblasts indicate that the efficiency of this type of nuclear staining is about 2 to 3 dye molecules incorporated per 100 nucleotide pairs. Quantitative data on fading of acriflavin fluorescence in stained fibroblasts indicate that fading is negligible in the time required to record the microscope images.


Does heparin occur in mucosal mast cells of the rat small intestine? Microspectrophotometric analyses combined with model experiments using polyacrylamide films containing different types of purified glycosaminoglycan or glycosaminoglycan and basic protein, have been applied to investigate the chemical characteristics of the glycosaminoglycan moiety present in rat mucosal mast cell granules. The results obtained with these histochemical techniques present evidence of the absence of significant amounts of heparin and the presence of lower sulfated glycosaminoglycan(s) in the granules of these cells.


[Alpha fetoprotein and neonatal jaundice. Contribution to the study of a physiopathologic mechanism]. The increase in the maternal plasma A.F.P. level is due to an hypoxia of the foetus. The prospective study of 851 single pregnancies shows that there is a significant rise in the A.F.P. levels during the last days of the pregnancy if the babies are going to exhibit a so-called physiological jaundice at birth: the decrease of the A.F.P. levels in these cases is four times slower than in normal cases. The prospective study of another group of 404 pregnancies gave the same results for the A.F.P. level of the blood of the umbilical cord. Statistical analysis showed that the pathological conditions capable of increasing the A.F.P. levels are related to neo-natal jaundice. The neo-natal jaundice may be due to a factor of foetal hypoxia capable of inducing an over stimulation of the foetal erythropoiesis. The results of this mechanism would be a quantitative disequilibrium between an increased hemolysis and a reduced bilirubine fixation capacity during the neo-natal period.


[Orange colored cells in the amniotic fluid. Statistical evaluation of 504 samples]. The authors have made a statistical evaluation of 504 estimations of the percentage of orange coloured cells in the liquor amnii. They have been able to draw up a curve of the means which confirms the value of this method. Orange coloured cells of fetal origin first appear after about 28 to 30 weeks. The rise in the percentage which is gradually progressive between the 30th and 37th weeks becomes sharp at the 38th week. The gradients of the curve of the means between the 37th and 38th week is comparable to that of the ratio lecithin/sphingomyelin. It is different from that of creatinaemia which is steeper. False-positive results are rare: at 2 p. 100 they seem to correspond to an advance in fetal maturation which may be spontaneous or may be brought about by treatment. False-negative results are commomer: 13 p. 100. The percentage of orange coloured cells should be viewed in conjunction with other tests for maturity. The technique is simple but the results may be vitiated by the minutae of preservation, of taking or of staining the cells which are described. Orange coloured cells maintain their value in pathological pregnancies and in particular in those where the fetuses are small for dates, for the percentage of orange coloured cells agrees more with the age of the fetus than with its weight.


Variable opacity of glycogen in routine electron micrographs. Glycogen in nerve terminals from the reticular zone of frog muscle was identified by transmission electron microscopy of both periodic acid-thiosemicarbazide-silverproteinate treated and UAc-PbCi-stained serial sections. A variable appearance of glycogen in the uranylacetate-lead citrate-stained nerve terminals was seen and is related to the preparative procedure. The study indicates the necessity of cytochemical identification for the assessment of glycogen organization in cells.


Rapid purification of peanut agglutinin by sialic acid-less fetuin-sepharose column. A simple purification of peanut agglutinin (PNL) with a sialic acid-less fetuin-Sepharose column is described. Immunofluorescence studies show PNL to be a suitable surface marker of potential use in the study of subpopulations of thymocytes and T cells.


A simple spot technique for thin layer immunoassays (TIA) on plastic surfaces. A simple and sensitive technique for visualization of antigen--antibody reactions is described. The property of many antigens to become adsorbed firmly on to a hydrophobic polystyrene surface while retaining their serological reactivity is taken advantage of. On a surface with adsorbed antigen the corresponding immune serum is applied spot-wise. The antigen--antibody reaction areas on the surface are characterized by a distinct hydrophilic condensation pattern when exposed to water vapour. The results obtained by the immunoassay technique described can be reproduced with great accuracy. The method is well suited for quantitative determination of a wide range of antigens as well as their corresponding antibodies. Antigen concentrations of 0.2--0.8 mg/l and antibody concentrations about 1 mg/l can be detected. By employing an antiimmunoglobulin serum subsequent to the primary antigen--antibody reaction, an increase in sensitivity can be obtained.


Leukocyte adherence inhibition: an automated microassay demonstrating specific antigen recognition and blocking activity in two murine tumor systems. This communication describes an automated micro-adherence inhibition assay. Tumor-specific immunity was demonstrated with B16 melanoma and MCA-38 colon adenocarcinoma, both of which are syngeneic to the same strain of mouse (C57B16/J). Abrogation of the leukocyte adherence inhibition (LAI) response of sensitized leukocytes has been demonstrated in the MCA-38 tumor system by the addition of serum from mice bearing MCA-38 progressively growing tumors, a property not present in normal serum. The sensitivity of the system has also been demonstrated by showing that LAI will change prior to a tumor becoming palpable. This microassay has the advantage of being simple, rapid and reproducible, and involves the use of minimal quantities of antigenic preparations and leukocytes, and in addition is amenable to rigorous statistical analysis.


Interaction between soluble immune complexes and glass-fiber filters. The two components of soluble antigen--antibody complexes, at the antigen excess, exhibit an increase in their binding ability to glass-fiber filters. This is demonstrated in two systems. In the 125I-BSA--anti-BSA system the proportion of 125I-BSA bound to the filter is markedly increased in the presence of anti-BSA antibodies. More than 80% of the antibody bound BSA can be removed by passage through the filter. In the other system, mouse gamma globulin (MGG)--125I-anti-MGG, the proportion of antibody bound to the filter increases with the increase in antigen concentration, whilst the presence of another, non-related, gamma globulin has little effect on the binding. The possible mechanisms for the binding of soluble complexes to the glass fibers are suggested and discussed.


A scanning electron microscopic and x-ray microanalytic study of cell surface material during amphibian neurulation. Treatment with lanthanum (La3+) after fixation in phosphate (PO4-3)-buffered glutaraldehyde results in the deposition of a cell surface material (CSM) primarily on the developing urodele amphibian neural axis. X-ray probe microanalysis indicates that calcium (CA2+) levels are considerably higher in the neural fold region. La3+ displaces Ca2+ from negatively-charged moieties on biological membranes. Once bound, La3+ likely interacts with residual phosphate(s) resulting in deposition of CSM. Elemental X-ray microanalysis shows CSM contains mostly lanthanum and phosphorus. The high level of regional La3+ binding is correlated with inherently greater Ca2+ levels in the developing neural axis.


Computer-assisted analysis of particulate axoplasmic flow in organized CNS tissue cultures. Using organized nervous system tissue cultures of embryonic mouse spinal cord, time-lapse motion pictures of intra-axonal particle movements were made using Nomarski optics. The paths of 272 particles were followed and analyzed using digital computer methods. Particles had a bidirectional saltatory motility with net anterograde and retrograde velocities which were shown to be the same (about 1 micron per second). When anterograde moving and retrograde moving particles were analyzed separately for anterograde and retrograde saltations, the velocities of saltation in the direction of net movement were found to be 1.5 times greater than saltations against the direction of net movement. These differences were statistically significant. In addition, it was shown that there were regions within the axon where variations in particle motion were similar from particle to particle as they passed, and that more disturbed motion was found to occur near intra-axonal objects which were judged to be mitochondria. No decrease of particle velocity was noted in regions away from mitochondria nor was in increase in velocity noted near them. Computer-drawn particle plots illustrate the various facets of particle motion.


Studies of cellular sensitization to myelin antigens in multiple sclerosis. Dissociation of MIF and LBT production in response to a peptide encephalitogenic in rhesus monkeys. The macrophage migration inhibitory factor (MIF) assay and the lymphoblastic transformation (LBT) technique were utilized simultaneously to measure immune responses to peptide Y, the 17 amino acid C-terminal fragment of basic myelin protein, in patients with multiple sclerosis (MS). Ten normals and 67 MS patients from the Montreal Neurological Hospital and affiliated institutions were examined. A prospective attempt was made to correlate the measured responses with phasic clinical activity of the disease. The LBT results indicate some degree of cellular sensitization to peptide Y which parallels the clinical course of the illness, and resembles earlier positive findings obtained with the whole basic myelin protein molecule. These findings, however, are in contrast to a negative MIF response to the Y peptide used in the present study and further contrast the positive MIF results obtained earlier using the whole protein. It is not evident from the results of the present study whether sensitization may be of any pathogenetic significance, but the findings show that differing portions of the basic myelin protein molecule may selectively stimulate specific lymphokine elaboration by sensitized lymphocytes.


HLA-typing and lymphocyte population studies in patients with multiple sclerosis. Compared to a control population, the frequency of HLA-B7 was increased in a group of 46 patients with a chronic progressive form of multiple sclerosis (MS) (RR = 2.9, P = 0.01). Significance of the association was lost after correction for first order error. Results on serotyping for "B-cell DW2" antigen are suggestive of an increased frequency of this antigen in chronic progressive MS patients (RR = 2.9, P = 0.01). The percentage of T-cells (E-rosette forming lymphocytes) in MS patients was lower (mean 46 +/- 17%, n = 34) than in a control group (mean 62 +/- 10%, n = 90). Only 5 of the 40 patients had high B-lymphocyte percentages, whereas no difference in B-cell counts was observed between the total group of patients and the control group. The T- and B-lymphocyte ratio in the MS patients was lower than in the control group. The 7 patients, with the lowest T/B-cell ratio were negative for "B-cell DW2" antigen. In 3 patients, the sum of T plus B-lymphocytes was far lower than 90%. At least 4 patients had in their serum antibodies against autologous lymphocytes as found by immunofluorescence tests. No correlation could be found between these serological and immunological data and clinical data on progression of the disease, the IgG in CSF and improvement after immunosuppressive treatment.


Enzyme and protein studies of demyelination in diabetes. Neuropathological evidence of demyelination was found in the brain and sciatic nerve of diabetic patients at autopsy. The activity of acid proteinase was somewhat increased in the white matter but decreased in the gray matter of diabetic patients. No increase was observed in the activity of neutral proteinase in diabetic white and gray matter. The activities of beta-glucuronidase and 2',3'-cyclic nucleotide-3'-phosphohydrolase (CNP) were of the same level as those of the controls. The activities of all 4 enzymes appeared to be increased in the diabetic nerve, with the possible exception of CNP which was measured from only 1 nerve. Furthermore, the amount of total protein was markedly decreased in diabetic peripheral myelin. The encephalitogenic basic protein of diabetic brain myelin was normal in the disc gel electrophoretic patterns of brain myelin proteins. However, the basic proteins of peripheral myelin were reduced in a number of diabetic patients. The present biochemical findings for diabetic white and gray matter were largely normal. Instead, the increased activities of at least the proteinases and beta-glucuronidase in diabetic peripheral nerve, together with the loss of basic proteins, indicate extensive biochemical damage of the peripheral nervous system in diabetes. They suggest that demyelination and other phenomena observed in diabetic peripheral nerve are not caused only by angiopathy and impaired circulation.


The beta 2-microglobulin content of the cerebrospinal fluid in neurological disease. Beta 2-Microglobulin levels in the CSF and serum of 125 neurological patients were determined. The concentration of beta 2-microglobulin in CSF and serum is approximately the same, ranging from 0.6--2.3 mg/l. No clear relationship could be found between the CSF beta 2-microglobulin level on the one hand and the total protein content and white cell count on the other. There was no clear relationship between the serum beta 2-microglobulin and CSF microglobulin content. An increase of beta 2-microglobulin in CSF was found most often in cases of serious infection of the meninges or of the central nervous system. An elevated beta 2-microglobulin content can occur as the only abnormal feature of the CSF.


A new procedure for examining Golgi impregnated neurons by light and electron microscopy. A procedure is described by which neurons impregnated by the rapid Golgi method can be de-impregnated and their fine structure studied in the electron microscope, Brains are fixed by perfusion with buffered solutions containing formaldehyde and glutaraldehyde, and pieces are then impregnated by a rapid Golgi method. Sections, 150-200 micron thick, are cut from the impregnated blocks and immersed in glycerol so that sections containing suitably impregnated neurons can be selected under the light microscope. Such sections are immersed in gold chloride followed by oxalic acid. The original impregnation deposit of silver chromate is then removed with sodium thiosulphate. This process of de-impregnation leaves the originally impregnated neurons still visible in the light microscope for they now contain a deposit of gold. In the electron microscope this deposit is apparent as fine particles that mark the profiles of the de-impregnated neurons, but the deposit is such that it does not interfere with the fine structure which is remarkably well preserved. Thus, the cytological details of the de-impregnated neurons can be discerned and their synaptic relationships can be determined.


2-(2-Aminoethylamino)-1,2-diphenylethanol derivatives, a new class of topical antiinflammatory agents. A number of analogues and derivatives of the title compound were synthesized and evaluated in a new test procedure used to detect topical antiinflammatory activity. Some general comments regarding observation on the structure-activity relationship of these compounds are made.


Ultrastructural effects of perfomic acid oxidation of epoxy-embedded tissue with and without early uranyl staining. Epoxy-embedded blocks of glutaradehyde and OSO4 fixed adenohypophyseal tissue were immersed in performic acid, rinsed and sectioned for electron microscopy. Sections exhibited a general image intensity equivalent to control sections but their stainability with both uranyl acetate and lead citrate stains was increased. It was concluded that osmium was not removed by performic acid oxidation. Membranes of endoplasmic reticulum and the Golgi apparatus were not visible, and other components (acidophil granules, in particular) were markedly distored in shape after oxidation. Such changes, however, were not evident in specimens previously exposed to uranyl acetate during ethanol dehydration.


The coelomocytes of the earthworm Lumbricus terrestris: morphology and phagocytic properties. The coelomocytes of Lumbricus terrestris have been classified and described, based on Wright's stained preparations and on living cells. The five major categories consist of basophils, acidophils, neutrophils, granulocytes and chloragogen cells. Both the acidophil and chloragogen cell groups contain two subgroups. Granulocytes also exhibit heterogeneity with respect to staining properties of granules. Some possess acidophilic granules, some basophilic granules, and others contain both types. Granules of acidophils have been observed to be occasionally excreted from the cells. All cell types, with the exception of chloragogen cells, produce pseudopodia and are capable of phagocytosis, a vital component of the earthworm's immune response.


Identification of an immunologically distinct papillomavirus from lesions of epidermodysplasia verruciformis. Virions isolated from warts of 2 siblings with epidermodysplasia verruciformis (EV), a rare disease characterized by the lifelong growth of warty skin tumors containing papova-like virions, were compared to isolates of human papillomavirus (HPV) from 3 pools of plantar and common hand warts. The length of relaxed, circular (form II) molecules of EV virion DNA approximated the length of HPV DNA but was 3.3% longer. Antisera prepared in rabbits against the 3 HPV pools coated and aggregated HPV in  immune electron microscopy (IEM) tests but did not react with EV virions. These antisera reacted at high titers in complement fixation (CF) tests with HPV and reacted only weakly in CF tests with EV virions. Rabbit antisera to EV virions coated and aggregated EV virions but reacted only weakly or not at all with HPV virions in IEM tests. These sera reacted in CF with EV virions only. The data indicated that virions from the EV patients represent an immunologically distinct papillomavirus.


Bleomycin: mammalian cell lethality and cellular basis of optimal schedule. Experiments on the dose- and time-dependent changes in the survival of Ehrlich ascites tumor cells exposed to bleomycin were done to determine a useful regimen for the effective inactivation of the tumor cells. The experimental results on time-dependent changes in survival of bleomycin-treated cells indicated that two phenomena were involved in the survival increase observed after single bleomycin treatment: 1) The bleomycin-treated cells were resistant to the second injection when the interval between the two successive doses was within 2 hours (induced resistance), and 2) the survival was increased as a function of time when the interval between treatments was prolonged (repair of potentially lethal damage). The effect of the fractionated treatments was then investigated for the regimen that minimized the induced resistance and repair of potentially lethal damage. The results indicated that administration of a lower dose of bleomycin at shorter intervals was more effective than other fractionation schedules, and, on the basis of the same dose rate, the continuous infusion regimen was more effective than fractionation regimens for the sterilization of Ehrlich ascites tumor cells.


Effect of dibutyryl cyclic adenosine 5'-monophosphate on protein synthesis in L5178Y cells after hyperthermia. Protein synthesis in the L5178Y mouse lymphoma cell line was stimulated after short-duration heat shock by the addition of dibutyryl cyclic AMP. The stimulation was concentration-dependent (range, 0.01-1.0 mM) and was enhanced by preincubation of the cells in the presence of the cyclic nucleotide before the heat shock. Actinomycin, which blocks the recovery of protein synthesis activity after heat shock, did not block the cyclic AMP stimulation.


Differential synthesis of mammalian type C viral gene products in infected cells. Radioimmunological techniques were applied to the quantitation of the translational products of the gag, pol, and env genes of mammalian type C viruses. Analysis of the viral proteins associated with simian sarcoma-associated virus (SSA V) and SSA V-infected cells revealed in each that the level of reverse transcriptase was less than 1% of that of the major viral structural protein, p30. The rate of intracellular degradation of reverse transcriptase in SSA V-infected cells was found to be no greater than that of several viral structural proteins, indicating that the lower levels of viral enzyme resulted from its decreased synthesis. By screening individual cells infected at limiting SSA V dilution, it was possible to isolate a clone (clone 16), which demonstrated levels of viral p12, p30, and gp70 similar to those found in wild-type SSA V-infected cells, and which released noninfectious virions in large quantity. The noninfectious virions and clone 16 cells were shown to lack immunologically or enzymologically detectable reverse transcriptase. With serial passage of clone 16 cells, reverse transcriptase activity became spontaneously detectable in tissue culture fluids, concomitant with the appearance of infectious virus. The reverse transcriptase associated with this virus was indistinguishable from SSA V polymerase, indicating that the genetic alteration restricting SSA V pol gene expression in clone 16 cells was reversible. These results further demonstrate the strict requirement of reverse transcriptase for establishment of type C virus infection. Possible mechanisms to account for the patterns of type C viral gene expression detected in SSA V-infected cells are discussed.


Urethral pressure profilometry: assessment of urethral function by combined intraurethral pressure and electromyographic recording. The technique of combined electromyographic and carbon dioxide urethral pressure profilometry is described and analyzed. The low viscous drag of carbon dioxide permits the use of catheter infusion rates above the critical threshold, where the compressibility of the gas does not influence the accuracy of the pressure recording. Clinical urethral function studies with this technique in benign prostatic hypertrophy, stress incontinence and traumatic spinal cord injury have documented the diagnostic value of the test. Furthermore, the impact of surgical or pharmacological treatment upon urethral function has been delineated.


How to set up a laboratory for investigation of neuromuscular dysfunction of the lower urinary tract. Neurourodynamics laboratories require the organization of sophisticated equipment for the individual evaluation of detrusor and urethral function. This equipment and these procedures provide greatest benefit to the patient when associated with direct physician participation. These procedures result in detection of a profile of test findings common to individual diseases of the lower urinary tract.


Dramatic palliation for painful, fixed bladder squamous cell carcinoma with 5-fluorouracil infusion. A case of an exquisitely painful squamous cell carcinoma of the bladder is reported. The pain progressed and rectal obstruction developed during palliative radiotherapy. With less than 7 days of infusion chemotherapy the patient became totally free of pain, with subjective and objective relief of the rectal obstruction (avoiding colostomy).


Rectal invasion by benign prostatic hyperplasia. Rectal invasion by benign prostatic hyperplasia occurred subsequent to an open transrectal biopsy of the prostate gland. Frequent transrectal resections of the prostate controlled this unusual lesion initially. However, massive intrarectal regrowth of this clinically malignant but histologically proved benign prostatic enlargement required eventual pelvic exenteration.


Telemedicine in critical care: an experiment in health care delivery. We hypothesized that telemedicine -- medicine practiced from a distance using telecommunications -- can solve some problems related to the scarcity and maldistribution of specialists in critical care medicine. Using a two-way audiovisual link between a small private hospital and a large university medical center, we have provided daily consultations by an intensivist to patients in the small institution. During the first 175 days of the project we found: 1) regular consultations in critical care can be provided using the audiovisual link; 2) current technology is adequate but expensive; 3) telemedicine consultations can be made acceptable to users and providers; 4) telemedicine can be a valuable educational resource; 5) telemedicine can influence the process and probably the outcome of patient care; 6) the audiovisual link is superior to the telephone for these consultations; and 7) telemedicine can serve as an important link between a small hospital and a large medical center favorably influencing the quality of care in the critical care unit of the small hospital.


[Relations between cell-bound lipids and disodium cromoglycate (author's transl)]. Disodium cromoglycate binds in vitro and in vivo to lipids in white cells. Smears of cells from lymphocyte cultures and from bone marrow aspirates treated with DNCG and subsequently stained with pseudoisocyanine show a characteristic green fluorescence (515 nm) of membrane- and intracellular-lipids. It is suggested that the mode of action of DNCG in the prophylaxis of bronchial asthma could be the binding of DNCG to membrane lipids. This binding might block the IgE-mediated reaction on the surface of mast cells which otherwise would lead to degranulation and release of vasoactive substances.


[Effect of chronic exposure to carbon monoxide on the biochemical composition of human blood]. Protein, lipid and carbohydrate metabolism of men was investigated during their prolonged exposure to a closed atmosphere containing carbon monoxide at a concentration of 20-10 mg/m3. A 30-day exposure of test subjects to carbon monoxide at a concentration of 20-15 mg/m3 caused an increase of albumin, beta-globulins, total lipids, cholesterol, beta-lipoproteins in the serum and a decrease of the sugar concentration in the blood. A 90-day exposure of test subjects to carbon monoxide at a concentration of 10 mg/m3 did not bring about significant changes in the above parameters. The provocative test for glucose tolerance showed no changes in the glycaemic regulation.


Clinical trial of live measles vaccine given alone and live vaccine preceded by killed vaccine. Fourth report to the medical research council by the measles sub-committee of the committee on development of vaccines and immunisation procedures. Follow-up of 5000 children given a single dose of live attenuated measles vaccine (Schwarz strain) when aged 10 months to 2 years shows a high level of protection in comparison with an unvaccinated group. This protection has been maintained for 12 years. Measles in vaccinated children was less severe as well as less frequent throughout the period. There is no evidence from the follow-up so far that a further injection of vaccine is needed; this has been confirmed by measles haemagglutination-inhibiting antibody estimations in a sample of the children.


Propranolol as an adjunct to the treatment of schizophrenia. Propranolol contributed usefully to the practical management of patients with chronic schizophrenia whose florid symptoms had not remitted with major tranquillisers. 14 patients who had received an average equivalent of 954 mg per day of chlorpromazine for 10 years were given, in addition, either propranolol or a placebo for 12 weeks. Both groups had improved by the twelfth week, but the propranolol group had improved significantly more.


Artificial sweeteners and human bladder cancer. A positive association between the use of artificial sweetners, particularly saccharin, and risk of bladder cancer in males has been observed in a case-control study of 480 men and 152 women in three Provinces in Canada. The risk ratio for ever versus never used is 1-6 for males (P=0-009, one-tailed test), and a significant dose-response relationship was obtained for both duration and frequency of use. The population attributable risk for males is estimated at 7%, though for diabetics, who have a similar risk ratio for artificial sweetner use as non-diabetics, the attributable risk is 33%.