Hormone-sensitive lipase of adipose tissue. Some physiologic aspects of the mobilization and fate of free fatty acids are reviewed. The molecular mechanism of the activation of hormone-sensitive lipase in adipose tissue is then discussed. Recent evidence established that hormone-sensitive lipase, concerned with fat mobilization, is both functionally and immunochemically distinct from lipoprotein lipase, concerned with uptake of plasma triglycerides. Lipoprotein lipase activity is not altered by cyclic AMP-dependent protein kinase. The latter enzyme enhances not only triglyceride hydrolase but also monoglyceride, diglyceride and cholesterol ester hydrolase activities in chicken adipose tissue. Finally, it is shown that the activation of all four acyl hydrolases is reversible, the deactivation being magnesium-dependent. Protein phosphatase fractions from heart and liver active against phosphorylase a can reversibly deactivate adipose tissue hormone-sensitive lipase, implying a low degree of substrate specificity for lipase phosphatase.


The use of psychotropic drugs in other painful conditions. Many studies have suggested that psychotropic drugs may be valuable in the managment of chronic painful conditions and of the pain due to neoplastic disease. An opiate-sparing effect has been postulated. Most of the studies are, unfortunately, uncontrolled. The question arises as to whether psychotropic drugs, in addition to allaying anxiety and depression, alter pain threshold or the appreciation of pain.


The measurement of pain in terminal carcinoma. 1. A method is described for comparing narcotic analgesics and adjuvants in patients with terminal cancer. 2. A randomized controlled trial of orally administered diamorphine and morphine is reported to illustrate the method. 3. It is concluded that there is no over-all clinical difference between diamorphine and morphine when administered by mouth every four hours at individually optimized doses in association with cocaine and a phenothiazine. 4. The validity, reliability and sensitivity of the method are discussed.


Simultaneous use of bleomycin and radiotherapy in malignant tumours of the head and neck. Twenty cases originally diagnosed as epidermoid carcinoma were observed with a view to determining whether a combination of radiation and intravenous bleomycin promoted regression of the tumours and if so what effect this had on survival. Signs of local toxicity with this combination were also looked for. Five cases yielded what was considered to be an excellent response, nine cases gave a good response and six cases had a poor response. It is recommended that future studies be carried out in epidermoid carcinoma with this combination of radiotherapy and bleomycin.


HLA-A1, B8-phenotype association and HBs antigenemia evolution in 440 hemodialyzed patients. HBs antigen (HBsAg) has been followed up every month in 440 hemodialyzed patients, typed for 26 HLA alleles of the A and B loci. An abnormally high rate of the HL-A-A1, B8 association (18.6%) was found in the group of patients able to eliminate HBsAg, when compared with the normal French population (5.05%, p less than 10(-4), and with the group of patients unable to eliminate HBsAg (7.0%,  less than 0.01). Chronic aggressive hepatitis was only found in the latter. This high frequency of the HLA-A1, B8 association has also been found in patients with seronegative active chronic hepatitis and suggests that this phenotype might be associated with high immune response against HBsAg.


Etiopathogenetic studies in a patient with Whipple's disease. A patient is presented with Whipple's disease. Before treatment, Haemophilus influenzae type e, sensitive to tetracycline was cultured from multiple small intestinal biopsies. This isolated micro-organism was structurally similar to the one observed in the tissue. All further culture experiments during and after treatment proved negative except for one biopsy from which a tetracycline-resistant H. influenzae type-e mutant was isolated. The immunological disturbances, mainly characterized by cutaneous anergy, in absence of major humoral or in vitro lymphocytic impairment, regressed during treatment together with clinical remission of the disease. These findings are considered in favour of the secondary nature of the immunological abnormalities.


A case report of primary hepatic carcinoma with prolonged HB virus infection and mono-clonal gammopathy. A case of alpha-feto protein (AFP) positive hepatic cell carcinoma has been presented in which the appearance of a mono-clonal gammopathy can be explained by persistent infection with HB virus (HBV). The patient, a fifty year old male, developed acute jaundice with hepatitis in July, 1952, and died of an hepatic cell carcinoma in December, 1973. The interest of this case lies in the fact that during the seven months prior to be patient's demise, a mono-clonal gammopathy of IgG (L type), together with positive titeres for HBs Ag (adr) and AFP, was demonstrated.


Clinicopathological studies of minute hepatocellular carcinoma. Analysis of 20 cases, including 4 with hepatic resection. Sixteen necropsies and 4 cases of hepatic resection in which the liver had a solitary hepatocellular carcinoma smaller than 4.5 cm, or a few tumor nodules smaller than 3.5 cm, have been analyzed. Clinically, these patients presented with signs and symptoms compatible with cirrhosis and, of the 16 autopsy cases only 2 had been diagnosed correctly. In all but 4 cases, the noncancerous parenchyma showed advanced cirrhosis of the mixed type, with irregularly sized multilobular nodules and thin strands of stroma, different from typical alcoholic cirrhosis. The primary lesion was grossly encapsulated in the majority, suggesting a slow, expanding growth. Histologically, most primaries were relatively well differentiated. Serum alpha-fetoprotein was generally low, and it served as the major diagnostic clue in only 5 cases. In patients with mildly abnormal alpha-fetoprotein levels, continuous monitoring seems important in order to detect a steady rise, the first warning for tumor growth.


Amidolytic assay of thrombin bound to alpha2-macroglobulin in plasma. A method for the determination in plasma of alpha2-macroglogulin-bound thrombin is described. Alpha2-Macroglobulin-bound thrombin is precipitated from plasma by 13% polyethyleneglycol, and its amidolytic activity is assayed by using the chromogenic substrate benzoyl-Phe-Val-Arg-p-nitroanilide (S 2160). After thrombin addition to plasma, only about 1.7% of the added thrombin activity was recovered in the alpha2-macro-globulin precipitate. It is suggested that the contribution of alpha2-macroglobulin to anti-thrombin activity of normal plasma is of little relevance.


[Pathomorphology of the human prostate gland]. The pathologic processes of the adult human prostatic gland, which consist of regressive and atrophic changes as well as of benign hyperplastic, metaplastic, dysplastic and carcinomatous proliferations, are analysed by light and electron microscopy. The examinations give rise to a new concept of the morphogenesis of each separate lesion. The results show the importance of the prostatic basal cell for the development of the special benign and malignant proliferations of the glandular epithelium. The histologically defined different alterations in the human prostatic gland are the results of the various differentiation possibilities of the basal cell.


The epidemiology of influenza.

The epidemiology of influenza. Although unpredictable, influenza outbreaks are known to occur in three patterns: pandemics every 30 to 40 years, with high excess mortality; epidemics much more frequently, with lower excess mortality; and usually mild sporadic outbreaks. The possibility of a swine-flu pandemic this winter, resembling that of 1918-20, is the result of a unique deviation in the epidemiology of this fascinating disease.


Encephalitogenic regions for the Lewis rat within the myelin basic protein. The sequence, phe lys asn ile val thr pro arg thr pro pro pro ser gln gly lys gly arg gly leu ser ser arg phe ser trp gly ala glu gly gln isolated from the peptic digestion of guinea pig myelin basicprotein is able to produce EAE in Lewis rats. The synthetic peptide phe lys phe gly gly arg asp ser arg, an analog of residues 154-162, is encephalitogenic in Lewis rats when B. pertussis is used as the adjuvant.


Further definition of the encephalitogenic region for guinea pigs. The effect on encephalitogenicity of using a serinyl substitution for the glutaminyl group in the peptide ser arg phe ser trp gly ala glu gly gln arg is reported. We conclude from these results that the function of the glutaminyl residue is assisting this peptide to produce allergic encephalomyelitis in guinea pigs is as a hydrogen donor-receptor.


In vitro tolerance induction to a thymus-dependent antigen with BALB/c and C57BL/6 lymph node cells: effect of tolerogen concentration and duration of exposure. Further studies on tolerance induction in vitro to bovine gamma globulin (BGG) in nonadherent BALB/c lymph node cells showed that it was dependent upon both the dose of tolerogen and the time of exposure. Nearly complete tolerance was achieved at a dose of 1.0 mg/ml and an incubation time of 12-18 hours. When C57BL/6 strain was used for comparison with the BALB/c strain because of its relative ease to become tolerant after in vivo injection of tolerogen, the rate of tolerance induction of its nonadherent lymph node cells was not different from that of BALB/c cells. Thus, in the absence of other host factors, the acquisition of tolerance by lymphocytes is gradual and may reflect the time required for a cell to reach a susceptible phase of the cell cycle and/or the activation of suppressor cells.


Histamine release from human leucocytes. A serum factor necessary for the induction of histamine release and desensitization by protein A. The ability of human leucocytes to release histamine on protein A treatment is lost when the cells are washed repeatedly. It is, however, possible to restore the sensitivity to protein A treatment by incubating the leucocytes in serum. Treatment of the cells with purified IgG does not restore the activity. The material responsible for the resensitization is eluted both in the second and the third protein peak when serum is chromatographed on Sephadex G-200, indicating the possible existence of several active factors. Material with low immunoglobulin content, but with retained capacity to resensitize leucocytes to release histamine on protein A treatment, was obtained by repeated chromatography of peak III material on the Sephadex G-200 column. Furthermore, material from the second and third peaks from Sephadex G-200 deprived of their IgG by passage through a protein A Sepharose or a DEAE-cellulose column had the same capacity to resensitize the leucocytes as unseparated material. When serum was separated by Pevikon block electrophoresis, most of the activity was detected in the alpha and beta regions but only little in the gamma region. The serum fractionations indicate that neither IgG nor the other immunoglobulins are the factor(s) responsible for resensitizing the leucocytes to release histamine on protein A treatment. Beside being necessary for protein A-induced histamine release, the factor (or factors) is also essential for protein A-induced desensitization of human leucocytes.


Immunosuppression in murine malaria. I. Response to type III pneumococcal polysaccharide. Acute Plasmodium yoelii yoelii and chronic Plasmodium berghei malaria infections of CBA mice were accompanied by a reduced capacity to give an antibody response to type III pneumococcal polysaccharide (SIII). The depression of response initiated by acute malaria persisted for several weeks after recovery from clinical infection. During chronic infection, and at the peak of acute parasitaemia, virtually no response to SIII was detected. A substance which crossreacted serologically with SIII was found in blood cells of infected mice. The results suggest that antigen-specific, as well as non-specific, factors may contribute to the depression of the response to this antigen.


Surface immunoglobulin on rabbit lymphoid cells. IV. Ultrastructural labelling of A and B group allotypic determinants. Equal numbers of rabbit peripheral blood lymphocytes (PBLs) label for either a- or b-group immunoglobulin allotypes using a modification of the mixed antiglobulin reaction of Coombs adapted for electron microscopy. This technique which employs immunoferritin markers is consistently more sensitive than other methods used for labelling surface Ig on lymphocytes. Although approximately 75 per cent of PBLs label for either a- or b-group allotypes, the number of immunoferritin grains on the cell surface after labelling for b group allotypes is nearly twice as great as that observed after labelling for a group allotypes. We conclude that essentially all Ig-bearing blood lymphocytes of the rabbit express both heavy chains (a allotypes) and light chains (b allotypes) on their surface membranes, but that light chain allotypic determinants are more accessible or exposed than the heavy chain (Fd) allotypic determinants.


Evidence for the presence of choleragen receptor on the thymocyte-brain antigen molecule of mice. CBA thymocytes were subjected to cocapping experiments using AThy-1.2 and choleragen. The finding that both structures showed a common ligant-induced redistribution implies that choleragen receptors of the cell membrane are associated with the thymocyte-brain molecule. Furthermore, it was found that the thymocyte-brain antigenic determinants on the thymocyte were not blocked by prior choleragen treatment. This suggests a spatial distance between these structures on the molecule.


Cellular mechanisms of the resistance to the induction of immunological tolerance. Tolerance inducibility was examined in various strains of mice by injecting aggregate-free HGG, followed by challenge with aggregated HGG plus LPS and by assessing spleen PFC numbers. Marked differences were shown between C57BL/6 and DDD mice. The antibody response to HGG was totally suppressed in C57BL/6 mice injected with 0-1 to 1 mg of aggregate-free HGG whereas little suppresssion was observed in DDD mice. Cellular mechanisms of the resistance to tolerance induction in DDD mice were explored. On the injection of 1 mg of aggregate-free HGG, spleen cells were only partially tolerized (at minimum, 21 per cent of control level on 5th day) and thymus cells were suppressed to 40 per cent of the normal responsiveness. No suppression was observed in bone marrow cells. Macrophages do not seem to play a decisive role in the resistance to tolerance induction in DDD mice as the injection of biofiltered HGG or the pretreatment with carrageenan did not ease the resistance. From these data, it was suspected that the resistance to tolerance induction to HGG in DDD mice might be due to the raised threshold of T cells for tolerance.


Comparative study of three methods for detecting avian leukosis viruses. This investigation was designed to compare detection limits for avian leukosis viruses after infection of chicken fibroblasts with decimal dilutions of Rous-associated virus type 1 (RAV-1). At 5, 9, 14, and 19 days postinfection, cells were examined for group-specific (gs) antigens by microtiter complement-fixation (CF) tests for avian leukosis viruses and by radioimmunoassays (RIA) for the major gs antigen having a molecular weight of 27,000 (p27). Culture fluids, collected at the same time periods, were also assayed for reverse transcriptase activities. We found that minimally infected cultures expressed virus proteins within 9 days postinfection regardless of method used. Although p27 RIA was consistently more sensitive than CF or reverse transcriptase assays, sensitivity was only two- to fivefold greater when concentrated suspensions of RAV-0, RAV-1, and RAV-2 were compared. In terms of infectious units, the lowest detectable virus titer was 6 X 10(3) infectious units as determined by RIA end point dilutions. However, our results led us to conclude that when concentrated cell extracts are tested with hamster antiserum, CF is adequate for detecting infection.


Common antigen of Mycobacterium leprae, M. lepraemurium, M. avium, and M. fortuitum in comparative studies using two different types of antisera. No. 21 mycobacterial antigens of Mycobacterium lepraemurium, M. avium, M. fortuitum, and M. leprae were compared in crossed immunoelectrophoresis using two different antibody sources, a serum pool from lepromatous leprosy patients (LSII) and a rabbit anti-M. smegmatis antiserum. M. lepraemurium, like M. avium, was found to contain the 21 A and 21 C determinants. M. fortuitum contained in addition a new type of determinant, 21 D.M. leprae antigen no. 21 carried the A as well as the B dertminants, the latter found so far only in the leprosy bacillus. The separate taxonomic position of M. leprae, suggested by earlier studies of the no. 21 antigen, is further supported by the present results, which also demonstrate the potential use of submolecular heterogeneity for such investigations.


Induction of rat homocytotropic antibody against streptokinase-streptodornase (SK-SD) and specific neutralization of the antibody by low molecular weight component of SK-SD. Homocytotrophic antibodies (HCA) were produced in rats by primary injection of streptokinase-streptodornase (SK-SD) after the antigen was absorbed on alminum hydroxide gel. The properties of rat SK-SD-specific HCA were qualitatively similar to SK-SD skin-sensitizing antibodies found in sera of human beings, namely, homocytotropic activity (48 h homologous PCA reaction), heat lability at 56 degrees C, and inactivation by reduction with 2-mercaptoethanol followed by alkylation with iodoacetamide. In neutralization tests, D-SK-SD, a low molecular hapten-like substance contained in SK-SD preparation, absorbed the antibody against ND-SK-SD antigen-specifically. The neutralizing activity mostly resided in fraction 1 of D-SK-SD separated through DEAE-Sephadex A-25 ion-exchange column, opposing the inhibitory activity of fraction 3 on lymphocyte transformation. The possible implication of these findings in the immunological treatment of streptococcal disorders is discussed.


Modulation of antigen-induced histamine release from bovine granulocytes by several anti-allergic agents. Sensitised bovine granulocytes release histamine when exposed to antigen. Several anti-allergic agents, some previously shown to be active in cattle, were tested to investigate their modulation of this histamine release process. Diethylcarbamazine citrate potentiated release at low concentrations and inhibited at high concentrations. Sodium meclofenamate and PR-D-92-EA were potent inhibitors. Acetylsalicylic acid and ICI 74,917 inhibited at high concentrations. Disodium cromoglycate was relatively ineffective, although it potentiated histamine release at low concentrations.


Release of histamine and formation of prostaglandins in human lung tissue and rat mast cells. Release of histamine and prostaglandins (PGs) from human lung tissue and rat mast cells was investigated. Passively sensitized lung fragments released PGE, PGF2alpha and the 15-ketodihydro metabolites into the media with time alone. Antigen challenge liberated 20% tissue histamine and there was a twofold increase in PGs. Twice as much PGF2alpha as PGE was found. beta-Adrenergic agonists inhibited the anaphylactic release of mediators and this action was blocked by propranolol. Both PGF2alpha and PGE were consistently found in the mast cell media, demonstrating that mast cells can synthesize PGs. Anaphylaxis induced a marked liberation of histamine but not of PGs. The results indicate that the release of histamine may precede the major release of PGs and suggest that the bulk of PGs may derive from cells in the proximity of the mast cells.


Subcellular localization of monoamine oxidase in bovine thalamus tissue using immunoferritin conjugates. A combination of discontinuous sucrose gradient analysis and polyacrylamide electrophoresis was used to isolate one of the multiple forms of monoamine oxidase (MAO) from bovine thalamus. This substance (the principle form and the most anodic of the five MAO forms observed) was used as a basis for an immunoferritin-electron microscope approach to determine the subcellular localization of MAO in thalamus. This form of MAO, as well as antigenically related forms, was found to reside mainly on the outer mitochondrial membrane. In addition, the action of SDS on solubilization and interconversion of MAO forms was studied and found to be dependent on the concentration and time of reaction of SDS with thalamus tissue.


The value of an assessment of erythema and increase in thickness of the skin reaction for a full appreciation of the nature of delayed hypersensitivity in the guinea pig. Delayed type skin reactions in guinea pigs have been assessed by measuring three parameters: increase in skin thickness, diameter of erythema and intensity of erythema. Some groups of animals were immunized with different protein antigens in Freund's complete adjuvant with or without cyclophosphamide (CY) pretreatment; others received a single high dose of antigen intravenously at the time of immunization. The results emphasize the importance of measuring all three parameters for several days after skin testing. The intensity or erythema was found to be an especially useful parameter for assessing CY-induced modification of delayed hypersensitivity (DH) reactions. The increase in skin thickness, which can be measured objectively, was also valuable as both immediate and DH reactions are characterized by induration. The diameter of erythema could only be measured accurately for a short time after skin testing. Furthermore, the effects of intravenous antigen and CY pretreatment were not reflected by that parameter.


Inhibition of antigen-induced histamine release by beta-adrenergic stimulants in vivo. IgE-mediated histamine release was studied using the method of passive peritoneal anaphylaxis (PPA) in the rat. Some beta-adrenergic stimulants markedly inhibited this reaction in vivo, the order of potency (ED50 microng/kg i.v.) of agents tested being fenoterol (6), salbutamol (40) and isoproterenol (94). Higher activity against the simultaneously measured dye extravasation suggested a dual effect of the drugs on both the cellular (inhibition of histamine release) and the vascular level. The order of potency in modifying vascular injury was, however, reversed, isoproterenol and not fenoterol being relatively more active here, as could be shown by further experiments. Inhibition of histamine release is discussed with respect to (a) methodical requirements and (b) the suggestion that beta2-receptor stimulants (fenoterol, salbutamol) are more selective than isoproterenol.


Comparison of murine delayed hypersensitivity reactions elicited with particle-associated versus soluble human gamma-globulin. Footpad reactions elicited in DHS mice using human gamma-globulin (HGG)-coated polystyrene latex particles and soluble HGG (sHGG) were compared. Both the magnitude and the persistence of DHS lesions produced by HGG-latex were considerably greater; at 24, 48 and 72 h after challenge, the level of reactivity induced by HGG-latex was 24, 41 and 71% higher, respectively, than those elicited with sHGG. Early nonspecific swelling following injection of HGG-latex in footpads of normal mice was negligible by 24 h, whereas Arthus-responsive animals did not return to control levels until 48 h. The possible advantages and limitations of using particle-associated protein to elicit DHS reactions are discussed.


Suppressor T cell product, which depresses the passive transfer of contact sensitivity, shares epitope(s) with the major histocompatibility complex. Specific suppressor T cell product appears in the supernatant of cultures of lymph node cells of mice injected with picryl sulphonic acid and then painted with picryl chloride. It is detected by its ability to depress the passive transfer of contact sensitivity by immune cells incubated in it. This communication shows that the product can be absorbed by antisera prepared against the major histocompatibility locus but not by antisera prepared against mouse immunoglobulin.


Characterization of EBV-genome negative "null" and "T" cell lines derived from children with acute lymphoblastic leukemia and leukemic transformed non-Hodgkin lymphoma. Sixty-two explants from peripheral blood, bone marrow and cerebral fluid of children with acute lymphoblastic leukemia (ALL) and leukemic transformed non-Hodgkin lymphoma (NHL) were cultivated for at least 8 weeks. Although lymphatic cells persisted up to 16 weeks in tissue culture, no proliferation was observed in 54 cultures. From the remaining cultures, eight permanently growing cell lines were obtained. Five of these were EBNA (Epstein-Barr virus-specific nuclear antigen)-positive. Three, however, were ENBA-negative and lacked Epstein-Barr virus genomes. Two cell lines (KM-3 and SH-2) expressed neither B nor T cell characteristics. One line (JM) expressed T cell characteristics and complement receptors. The growing lymphatic cells represented leukemic cells, since the pattern of cytochemical staining and that of membrane receptors of lymphoblasts from the same donor prior to cultivation were identical. All leukemic cell lines were derived from patients in relapse. In contrast, no proliferation of leukemic cells occurred in explains from patients revealing the first manifestation of the disease. These results suggest enhanced growth potential of lymphoblasts resisting antileukemic therapy.


A comparative study of the localization of CEA and NCA2 in cancerous and normal gastrointestinal tissues. The histological localization of CEA in gastrointestinal tissues was re-evaluated after absorption of anti-CEA antiserum with NCA2, another normal antigen cross-reacting with CEA. This absorbed antiserum showed clearly the presence of CEA in colonic tumors and some non-cancerous colonic mucosae, obtained even from non-cancerous patients. In contrast, some of the gastric adenocarcinomas we studied were strained very weakly by absorbed anti-CEA antiserum, although non-absorbed antiserum (or serum absorbed with NCA alone) labelled them strongly. The same difference in reactivity between both antisera was observed in intestinal metaplasia. The localization of NCA2 was studied with a specific antiserum, previously absorbed with CEA and NCA. NCA2 was found to be a noromal cytoplasmic and mucus-associated antigen of gastrointestinal tissues. It was present also in fetal stomach and colon. Sections of gastric and colonic tumors as well as intestinal metaplasia of gastric mucosae always reacted brilliantly with absorbed anti-NCA2 antiserum.


Detection of SV40 T antigen with labelled antibodies: radioimmunoassay and autoradiography. SV40 T antigen, partially purified from SV80, a human cell line, has been used to develop a sensitive solid phase radioimmunoassay. In this assay SV40 T antigen was immobilized on microtiter plates, reacted with hamster anti-SV40 T antibody and subsequently with 14C-labelled anti-hamster IgG. The test detected approximately 20 ng of partially purified SV40 T antigen and is rapid and more convenient than the complement fixing reaction. An immunoautoradiographic method which permits the determination of the cellular localization of SV40 T antigen and which is possibly more sensitive than the indirect immunofluorescence method is also described.


Cellular and humoral anti-tumor immune responsiveness in chickens bearing tumors induced by avian sarcoma virus. A systematic comparison was undertaken of the respective abilities of normal chicken embryo fibroblast (CEF) cells, Rous sarcoma virus (RSV)- transformed CEF cells, avian Rous sarcoma (RS) tumor cells and murine RS cells to serve as targets and antigen donors in various assays for the detection of cellular and humoral anti-tumor immunity in chickens bearing tumors induced by Rous sarcoma virus. As measured by a cytotoxicity procedure, avian and murine RS cells were more susceptible to the killing effects of sensitized lymphocytes than were transformed CEF which in turn were more reactive than normal CEF. In contrast, sera from tumor-bearing animals were able to stain by indirect immunofluorescence only the avian RS and transformed CEF cell types. Extracts of both transformed CEF cells and avian RS cells but not normal CEF were equally effective as inhibitors of migration of peritoneal exudate cells derived from tumor-bearing animals. Transformed CEF were found to produce far higher quantities of transforming virus progeny than avian RS cells, although the latter were apparently albe to synthesize defective viral particles. These data indicate the significant variations which may occur under experimental conditions, depending on the types of assays and target cells employed.


Immunogenicity of solubilized tumor antigen extracted from P1798 murine lymphoma cells or isolated from tumor-bearer ascites fluid and reactivity with anti-thy-1.2 antiserum. Solubilized antigen was prepared from P1798 lymphoma cells by sonication, 3 M KCI extraction, or isolated from the ascites fluid of syngeneic tumor-bearing BALB/c mice. Antigen was detected and quantitated by its ability to block activity of anti-P1798 serum raised in syngeneic mice, as assayed by cytotoxic and indirect immunofluorescence tests. It was established that the reaction was immunologically specific as the P1798 antigen did not inhibit the binding to L1210 lymphoma cells of antisera raised against L1210 in syngeneic DBA/2 or allogeneic BALB/c mice. Vaccination of BALB/c mice with different subcellular fractions of sonicated antigen or with ascites fluid resulted in protection against a live P1798 challenge with results comparable to those obtained using iodoacetamide-modified tumor cells. Solubilized antigen prepared by each of the three methods eluted from a Bio-Gel A5m agarose column exclusively in an early peak that had a molecular weight estimated to be greater than 2 X 10(6). This column-fractionated antigen was shown to cross-react with antiserum raised against Thy-1.2 antigen, which is present on P1798 cells. The purified P1798 antigen sedimented at 200,000 g and was shown to protect syngeneic mice in immunoprophylactic tests.


Immunologic responses to a murine mammary adenocarcinoma. I. Passive transfer of immunity by sera from tumor-bearing mice. DBA/2 mice bearing a syngeneic mammary adenocarcioma T1699 produced high levels of tumor-specific antibody, detected by indirect immunofluorescence and subsequently identified as the IgG2a subclass. Tumor-bearer sera passively administered to normal recipients protected the animals from subsequent challenge with T1699 tumor cells butnot from challenge with a non-cross-reacting syngeneic tumor, SaD2 fibrosarcoma. Administration of sera prior to tumor challenge was found to be more effective than treatment after the challenge. The protective effect of sera appeared to parallel both antibody titers and appearance of concomitant immunity; however, sera absorbed with T1699 cells, with the indirect fluorescent antibody titers reduced more than 100-fold, conferred an almost identical level of protection. Immune suppression of serum recipients before serum transfer abolished the effect, suggesting that protection depended on a cellular immune response by the host in addition to the possible protective effect(s) of humoral antibody.


1-naphthyl acetate esterases in fluids and tissues of jaw cysts. The activity and electrophoretic mobility of 1-naphthyl acetate esterases in cystic fluids and cystic tissues of ameloblastomas, follicular and apical cysts were examined. The cystic fluids showed lower activities than sera but had very similar patterns on the electrophoretogram. The activity levels of the three kinds of cystic fluids were not statistically significantly different. The fluid esterases may have originated from serum but they were not produced by the cystic lining tissue. Ameloblastoma tissues showed the highest activity per wet weight and per mg protein of the three kinds of cyst lesions (P less than 0.05). On the electrophoretogram, the esterase-I activity constituted 41% of the total activity in ameloblastomas, whereas in follicular cysts and apical cysts the esterase-I activity constituted 32% and 24% of the total activity, respectively.


Electron microscopy of cartilage proteoglycans. The proteoglycans of cartilage are complex molecules in which chondroitin sulphate and keratan sulphate chains are covalently linked to a protein core, forming a polydisperse population of proteoglycan monomers. By interaction with hyaluronic acid and link proteins, the monomers form large macromolecular complexes. In vivo the proteoglycans mainly occur in such aggregates. In the electron microsope, the cartilaginous matrix can be seen to be made up of thin collagen fibrils and polygonal granules about 10-50 nm in diameter Addition of the polyvalent cationic dye Ruthenium Red to glutaraldehyde and osmium tetroxide fixatives yields a dense selective staining of the matrix granules. Following a short digestion of cartilage slices with either of the chondroitin sulphate-degrading enzymes hyaluronidase and chondroitinase or with the proteolytic enzyme papain, the matrix granules were few in number or completely absent and the proteoglycan content, measured as hexosamine, decreased by up to 90%. Similarly, extraction of the cartilage with 4 M guanidine-HCl removed all matrix granules and most of the proteoglycans. From these findings, it can be concluded that the matrix granules represent proteoglycans, most probably in aggregate form, and that Ruthenium Red staining may be used to study the distribution of these macromolecules in thin sections. As a complement to chemical studies on proteoglycan structure, it is also possible to observe and measure individual molecules in the electron microscope after spreading them into a monomolecular layer with cytochrome c. This technique has been applied in investigations on proteoglycans isolated from bovine nasal cartilage and other hyaline cartilages. The molecules in the monomer fractions appeared as an extended central core filament to which about 25--30 side-chain filaments were attached at various intervals. The core filament, averaging about 300 nm in length, was interpreted as representing the polysaccharide binding part of the protein core and the side-chain filaments, averaging about 45 nm in length, as representing the clusters of chondroitin sulphate chains. Statistical treatment of the collected data indicated that no distinct subpopulations existed within the monomer fractions. The electron microscopic results correlated well with chemical data for the corresponding fractions and together with recent observations on various aggregate fractions strongly support present concepts of proteoglycan structure.


Polyacrylamide films as a tool for investigating qualitative and quanitative aspects of the staining of glycosaminoglycans with basic dyes. With the introduction of model films of polyacrylamide gel into which purified glycosaminogly cans (GAGs) have been 'incorporated', the direct recording of metachromatic spectra with virtually no interference of the corresponding orthochromatic peaks has become possible. Because this model system yields situations comparable to those of stained sections under the microscope, it is well suited for investigating qualitative and quantitative aspects of histochemical staining procedures. Previous model experiments have shown that under aqueous conditions only minor differences can be observed between the metachromatic peaks of different GAGs complexed with a suitable dye (e.g. Toluidine Blue O, Thionin, Safranin O, Cresyl Violet, Cystal Violet). In non-aqueous media, such as glycerol and ethylene glycol, the complexes with Toluidine Blue O revealed a special pattern for heparin, having a metachromatic peak (517 nm) about 30 nm lower than that of all other GAGs. This observation has formed the basis of a method for the qualitative microspectrophotometric detection of heparin in situ which was worked out by combining model film experiments with microspectrophotometric data obtained from rat mast cells. Since only a limited number of cells in necessary for obtaining reliable data with this method, the presence of heparin in the cytoplasmic granules of normal human mast cells and basophilic granulocytes could thus be proved directly. Alcian Blue 8GX, another basic dye frequently use in GAG histochemistry, has also been investigated with polyacrylamide films. In contrast to the metachromatic dyes, the rate of staining with Alcian Blue depends to a large extent on the rate of penetration of the dye into the model films. The rate of penetration is also a phenomenon of great importance for dye binding in situ, where complex basic protein molecules may form a barrier for the Alcian Blue molecules. The model film studies performed so far have yielded conditions that provide maximal staining (up to an optimal level) and a linear relationship betweeen the concentration of GAG and the AB binding. The presence of basic protein, electrostatically bound to the GAG, was not found to influence either the rate of staining or the maximal amount of dye binding.


The Haematoxylin-basic Fuchsin-picric acid staining reaction as a test of myocardial viability in resuscitated and preserved hearts. In the field of the transplantation of organs, there is a great need for an in vitro test of viability which would confirm that the organ was capable of performing its normal in vivo functions. Such a test should ideally be simple, rapid and reproducible. Preliminary studies using the Haematoxylin-Basic Fuchsin-picric acid (HBFP) staining reaction to assess myocardial ischaemia in resuscitated and preserved hearts would suggest that this test meets many of the requirements of a viablility assay. The test has been employed in hearts which have been in a state of anoxic arrest for 30 min and then resuscitated and preserved as an autoperfusing heart-lung preparation. The 'positive' response after 30 min anoxic arrest reverts to a 'negative' response after 2 h myocardial perfusion. In hearts which have been preserved as an autoperfusing heart-lung preparation with no interim period of anoxic arrest the HBFP stain response remains 'negative' throughout, confirming satisfactory myocardial perfusion.


Histochemical localization of 3 beta-hydroxysteroid dehydrogenase in the testes of chicken-pheasant hybrids. Histochemical studies on the activity of 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD) in the testes of adult chicken-pheasant hybrids and domestic fowl of similar age were carried out using dehydroepiandrosterone as the substrate. The reaction for 3beta-HSD was positive in the interstitial tissue and negative within seminiferous tubules of domestic fowl. In chicken-pheasant hybrids, a strong positive reaction for 3 beta-HSD was observed in the interstital tissue and within the seminferous tubules suggesting that, in hybrid testes, both Leydig cells and cells of seminiferous tubules may be capable of steroid biosynthesis. Since the plasma testosterone levels in these birds were found to be extremely low, it is hypothesized that either the chicken-pheasant hybrids do not release testosterone in sufficient amounts in the circulation or the type of steroid produced by the 3 beta HSD-positive cells may be different from testosterone that is required for the maintenance of normal fertility and the development of secondary sexual characteristics.


The effect of sodium chloride on the extraction of DNA fragments during Feulgen acid hydrolysis. Feulgen acid hydrolysis was performed on ascites tumour cells labelled with radioactive DNA-precursors. The development of fragments of apurinic acid and the extraction of purines were studied by monitoring the variations in the extraction rate during the hydrolysis when sodium chloride was either present or absent from the hydrolysis solution. The changes in the rate of extraction of purines and the alterations in the initial retardation of the apurinic acid extracting process followed approximately the same pattern. The extractability of apurinic acid fragments during hydrolysis in 0.3 M HCl was found to be a maximum when the sodium chloride concentration was about 1 M. Sudden exchange experiments, in which acid was substituted for sodium chloride after various times of hydrolysis, revealed a successive shortening of the extractable fragments during the low acid concentration hydrolysis. The results strengthen the view that, during hydrolysis, apurinic acid is lost from the cells through a reaction whose form is determined, first, by an initial retardation of the depolymerization, second, by the maximum length at which fragments developed through the depolymerization become soluble and are lost by diffusion, and last, at low acid concentrations, by a mechanism whose influence is equivalent to the presence of bonds between the fragments and an unextractable stable structure.


Permanganate fixation demonstrates the monoamine-containing granular vesicles in the SIF cells but not in the adrenal medulla or mast cells. Three types of monoamine-storing cells were fixed with permanganates and analyzed ultrastructurally. The SIF cells contained the typical granulated vesicles characteristic of monoamines. On the other hand, both the catecholamine-storing cells in the adrenal medulla and the mast cells were devoid of a dense core in their monoamine vesicles. This is surprising, since permanganate is considered a reliable means of demonstrating monoamines at the fine structural level. The significance of this finding is discussed against the close relationship between SIF cells and cells of the adrenal medulla. Because permanganate as a fixative is widely used in monoamine fine structural cytochemistry, the present findings undoubtedly need further clarification.


Age-related changes in chromatin of liver cell nuclei of different ploidity. A comparison of the Feulgen hydrolysis curves and the chromatin compactness of the liver cell nuclei of young and old rats was made. It was found that the rate of DNA depurination and chromatin compactness are higher in the liver cell nuclei of old rats, both in di-and tetraploidal cells. The effect of fixation upon the course of the hydrolysis curves is discussed.


Oxygen-dependent inactivation of gramicidin S synthetase in Bacillus brevis. Incorporation of L-[14C]ornithine into gramicidin S by crude, unfractionated lysozyme extracts of Bacillus brevis ATCC 9999 was shown to represent the activity of the gramicidin synthetase complex. Frozen-thawed cells were the source of active extracts, but when cells were shaken in air at 37 degrees C, they rapidly lost activity in a first-order reaction with a half-life of 13 min. Protease inhibitors and inhibitors of energy metabolism had no effect on the inactivation process in frozen-thawed cells. Stabilization was achieved when the cells were shaken in nitrogen or helium instead of air. The addition of dithiothreitol produced a moderate degree of stabilization. The L-ornithine- and D-phenylalanine-activating activities of the gramicidin S synthetase complex were also lost during aeration of the cells. Crude cell-free extracts also lost activity when they were shaken in oxygen, but, in this case, inactivation was slower (half-life of 80 min). Nitrogen also stabilized these cell-free extracts.


Blood group activity of human sucrase from intestinal metaplasia. Sucrases were purified from human small intestine and from areas of intestinal metaplasia of the stomach mucosa surrounding stomach cancers. The kinetic constants and pH activity profiles of enzyme preparations from the two sources were similar. No blood group activity of sucrase was detectable in preparations from three cases of intestinal metaplasia, but preparations from two other cases showed activity like that of the small intestine. These results indicate that sucrase from areas of intestinal metaplasia has similar enzymatic properties to those of enzyme from the small intestine, but that the antigenic sugar moiety of the enzyme associated with blood group activity varies.


Rabbit globin mRNA: analysis of T1 RNAse digestion fragments. Rabbit globin complementary DNA made with RNA-dependent DNA polymerase (reverse transcriptase) was used as a template for in vitro synthesis of 32P-labeled RNA and deoxysubstituted RNA. The sequences of the nucleotides in most of the fragments resulting from combined ribonuclease T1 and alkaline phosphatase digestion have been determined. In addition, the 3' nearest neighbor was determined for several fragments resulting from digestion with T1 ribonuclease. The utility of the deoxysubstitution technique was demonstrated by the ease with which the sequences of pyrimidine-rich fragments could be determined. Many sequences thus determined were long enough to fit uniquely with the alpha- or beta-globin amino acid sequences. The positions of these fits were found to be clustered, leading us to believe that only certain regions of the complementary DNA are transcribed by Escherichia coli RNA polymerase. Other unique characteristics of RNA synthesis from a complementary DNA template include a high yield of free poly(A) and the fact that one must use low rather than high salt buffers to obtain transcripts of high molecular weight.


Methods for detection of hepatitis B surface antigen in paraffin sections of liver: a guideline for their use. Methods for the localisation of hepatitis B surface antigen (HBsAg) in paraffin sections of the liver include the detection of ground-glass hepatocytes and the use of Shikata's orcein stain, and of immunoperoxidase and immunofluorescent techniques. A comparative study of the different methods on 20 livers shows the orcein stain to be the method of choice for routine use. The Shikata stain is not only specific but is relatively inexpensive, easily performed, and stains out distinct cytoplasmic inclusions even in stored formalin-fixed livers, old paraffin blocks, and autolysed livers. Since HBsAg is irregularly distributed in the liver, adequate sampling is necessary to prevent false negative; when sufficient tissue is available at least five blocks should be examined before a case is labelled as HBsAg-negative.


Retinal projections in the native cat, Dasyurus viverrinus. Retinal projections were examined in the native cat, Dasyurus viverrinus using Fink-Heimer material and autoradiography. We found six regions in the brain which receive retinal projections. These are (1) the dorsal lateral geniculate nucleus (2) the ventral lateral geniculate nucleus (3) the lateral posterior nucleus (4) the pretectum (5) the superior colliculus, and (6) the accessory optic system. We did not examine the hypothalamus. The accessory optic system and the lateral posterior nucleus receive a contralateral retinal projection only and the other four regions receive a bilateral retinal projection. There is extensive binocular overlap in the dorsal lateral geniculate nucleus. On the side contralateral to an eye injection of 3H leucine our autoradiographs show four contralateral layers which fill most of the nucleus. Three of these layers, 3, 4 and 5, also receive input from the opsilateral eye. Layer 1 which lies adjacent to the optic tract receives only contralateral retinal input. Layer 2 receives a direct retinal input only from the ipsilateral eye. The ipsilateral projection to the dorsal lateral geniculate nucleus forms a fairly continuous patch which is not divided into separate layers. The ipsilateral retinal input is located in the dorsal part of the lateral geniculate nucleus. The ventral quarter of the nucleus only receives a contralateral retinal input and therefore represents the monocular part of the visual field.


Effect of potassium isobutyrate on absorption of immunoglobulins from colostrum by calves. Calves received either colostrum with 2.83 meq potassium isobutyrate per gram of gamma-globulin or colostrum with distilled water within 1 h after birth. Blood samples were taken at intervals during the first 72 h for determination of concentrations of gamma-globulin, immunoglobulin IgG and IgM. Calves fed colostrum with distilled water attained higher concentrations in serum of total gamma-globulin, immunolgobulin IgG and IgM. Holstein calves were more efficient than Ayrshire calves in absorbing total gamma-globulin, but differences between breeds were not significant for specific immunoglobulins. Efficiency of gamma-globulin absorption within 24 h was 35.7% for control calves and 24.7% for calves fed colostrum plus potassium isobutyrate. Potassium isobutyrate had a depressing effect on absorption of immunoglobulins by calves.


The role of ragweed pollen in autumnal asthma. Thirty-nine ragweed-allergic seasonal asthmatics were studied from 1972 to 1974. After quantitative skin tests, antigen E-induced leukocyte histamine release, quantitative inhalation bronchial challenge with ragweed extract to determine PD35 (provocation dose of allergen causing 35% decrease in specific airways conductance), and radioallergosorbent test (RAST) determinations were done, patients were paired based on PD35 values and randomly assigned to treatment or placebo groups, receiving either aqueous ragweed extract or placebo prior to the 1973 ragweed season. Treated patients received a mean cumulative dose of extract equivalent to 11.7 microng antigen E (4,180 protein nitrogen units [PNU]). Twenty-nine patients were followed through the ragweed season with daily symptom diaries and biweekly physician examinations. Severity of disease was not predictable by PD35 data, skin tests, leukocyte histamine release, or radioallergosorbent test (RAST) values. Although all patients were ragweed-allergic by objective tests, only 13/29 had asthma symptoms correlating with ragweed counts. Mold spore counts were related significantly to symptoms in some patients. Asthma and hay fever symptoms correlated significantly in 24/29 patients. This dose of immunotherapy caused no significant difference to be found in asthma or hay fever symptoms in treated versus placebo patients for the 1973 reporting period as determined by physician evaluations or daily symptom diaries. No patients showed significant improvement in PD35 values after treatment in 1973. Similar findings were obtained for a smaller group of patients followed through the 1974 ragweed season who received a mean dose of 31.2 microng antigen E (11,140 PNU). The failure of these patients to show a response to immunotherapy could be due to a combination of the relatively low dose of ragweed extract and their sensitivity to other allergens.


Pitting stain with soft contact lenses--Hydrocurve thin series. A survey of 85 of our patients (170 eyes) wearing relatively steep based curved Hydrocurve thin series hydrophilic lenses, shows that 32 percent of the eyes fit demonstrate a particular type of corneal staining (21 percent light, 6 percent moderate, and 5 percent heavy). We have designated this corneal epithelial staining as Pitting Stain. It occurs as concentrated, superficial, punctate staining that may coalesce and deepen into branching patterns with a whitish pitting appearance.


Incidence of central edema and peripheral staining as a function of contact lens fitting philosophy. A study of three hundred and fifty-two patients (700 eyes) was undertaken to determine the relationship (if any) between abnormal corneal physiology and methods of fitting hard contact lenses. This was divided into two categories of slit-lamp findings, i.e., central edema or central corneal clouding (ccc), and peripheral (3/9) staining. The categories of lens fitting methods were broken down into (a) alignment, (b) steep and (c) flat. The results and relationships are reported.


Commercial horseradish peroxidase degrades myelin encephalitogenic protein during coupling for immunohistochemical studies. Conjugates of myelin encephalitogenic basic protein (EP) and commercial horseradish peroxidase (HRP) have been used for immunohistochemical demonstrations of anti-EP antibody in animals with experimental allergic encephalomyelitis. We performed gel electrophoresis studies on EP-HRP conjugates prepared with glutaraldehyde and on mixtures of EP and HRP incubated without glutaraldehyde. The results show that under conditions of one-and two-step coupling HRP causes rapid loss of the native EP band, apparently due to EP degradation. The EP-HRP mixtures are not encephalitogenic in rabbits, or encephalitogenic activity is lost during processing. The immunohistochemical reactivity of conjugates, however, signals some preservation of antibody-combining sites. The mechanism of the HRP effect on EP is unknown. The possibilities of a contaminating proteinase or direct peroxidatic attack are suggested. Until this action of HRP can be overcome, the effect of coupling procedures on the biological activities of EP will be difficult to assess, and EP-HRP conjugates cannot be expected to reveal sites that may bind encephalitogenic portions of the EP molecule.


An improved acriflavine-Feulgen method.

An improved acriflavine-Feulgen method. The acriflavine-Feulgen method for the histochemical demonstration of deoxyribonucleic acid was modified by staining hydrolyzed cells with 0.01% acriflavine dissolved in 90% ethanol. This method offered the following advantages: (a) it simplified the preparation of the acriflavine-Feulgen reagent; (b) it left the cytoplasm essentially unstained while staining the nuclei bright green in hydrolyzed cells and left the cytoplasm and nuclei essentially unstained in unhydrolyzed cells; (c) it eliminated poorly defined reagents from the staining solutions. Because of these staining properties, this technique may be especially useful in the quantitative determination of deoxyribonucleic acid by cytofluorometry.


Further evidence in support of cell-surface-associated deoxyribonucleic acid in tumor cells: an autoradiographic study. Ascites sarcoma-180 cells, when stained with platinum-pyrimidine complexes as the sole electron dense stain, show distinct dense patches to granular appearance on the surface of the plasma membrane which has been suggested to be attributable to deoxyribonucleic acid. Swiss Webster mice, 4-5 weeks of age, weighing 24-26 g with 4 X 10(6) ascites sarcoma-180 cells when injected with 3 X 7.0 micronC of tritiated thymidine on day 5 of the tumor implant, show specific labeling on the plasma membrane surface. The photopositive silver grain distribution in both the light and electron microscope autoradiograms when followed from the nucleus outwards show a distinct peak over the nucleus and the plasma membrane. The quantity and origin and role of this surface-associated deoxyribonucleic acid is not clear.


Ultrastructural localization of acetylcholinesterase in cultured cells. II. Cycloheximide-treated embryo muscle. The acetylcholinesterase activity (AChE) of cultured chick embryo muscle fibers that remains after the cells have been treated with the protein synthesis inhibitor cycloheximide was examined with cytochemical stains and the electron microscope. AChE activity that decreased rapidly after addition of the inhibitor was associated with enzyme within the cells, and AChE activity that was relatively insensitive to the inhibitor was associated with AChE outside of the cells. The results support the view that there are at least two fractions of AChE in developing muscle fibers, one intracellular and labile, the other extracelullar and stable.


Neutralization kinetics studies with type SAT 2 foot-and-mouth disease virus strains. A comparison of homologous and heterologous rates of neutralization demonstrated that antigenic relationships of foot-and-mouth disease virus strains could be differentiated quantitatively by the kinetics of neutralization method described previously (Rwysed this way gave R values which were similar to those obtained with other neutralization test methods but which were generally smaller than those obtained with complement fixation test results. It was demonstrated that there were wide differences between the vaccine strains tested as demonstrated by R value relationships. An examination of r values, however, demonstrated that antisera to the Moz 1/70 strain were highly reactive with most of the virus strains from Central and Southern Africa. The selection of FMD virus strains with a wide serological range for vaccine production is discussed.


Specific restoration of delayed hypersensitivity by lymphoid tissue extracts. Mice lose demonstrable delayed hypersensitivity (DH) to DNFB, picryl chloride, or sheep red blood cells. Reconstitution of immune responsiveness can be accomplished by administration of cell-free lysates of spleens from mice with active DH to structurally related, but not to unrelated antigens. Peritoneal exudate cell lysates from mice with active DNFB-DH also restore DH to this antigen. Sera from sensitized mice, and sera and lymphoid tissue extracts from unsensitized mice are without activity. The restorative property of splenic lysates from DNFB-sensitized mice is unstable at 56 degrees C, not sedimented at 90,000 X G and inactivated by trypsin or magnesium ions. The presence of unexpressed, restorable DH may provide a biologic basis for the so called "transfer factor" phenomenon.


Specificity and structural analysis of a guinea pig transfer factor-like activity. A transfer factor-like activity was prepared by Sephadex G-25 chromatography of immune guinea pig leukocyte lysates. This isolated material leads to antigen-dependent migration inhibition and thymidine uptake by nonimmune lymphoid cells. Tests of the "transfer factor" from guinea pigs immunized to either ovalbumin or bovine gamma-globulin demonstrated the donor specificity of the in vitro activity. The activity is susceptible to heat (56 degrees C), alkali (0.5 M sodium hydroxide), pronase, and phosphodiesterase. The pronase susceptibility is blocked by traysylol, a protease inhibitor; the phosphodiesterase susceptibility is not bocked by traysylol. The guinea pig factor was purified further by alkaline phosphatase treatment. Sephadex G-25 chromatography, and DEAE-cellulose chromatography. The final product, active in vitro, represents about 0.03% of the cellular material absorbing 260 nm light, and contains polymerized amines and phosphate. Gel electrophoresis of the fluram-reactive components suggests a limited heterogeneity of the DEAE-cellulose-purified material. These data are consistent with the active "transfer factor" molecule including both peptide and phosphate-containing components.


A spontaneously occurring complex of beta2-microglobulin and a fragment of gamma-chain of IgG: isolation from the urine of a patient with plasma cell leukemia and characterization. A complex of a fragment of gamma-chain of IgG and beta2-microglobulin (beta2M) was isolated from the urine of a patient with plasma cell leukemia. The approximate m.w. of the gamma-fragment was 19,000 and this gamma-fragment was found to be associated with beta2M noncovalently. The complex could be dissociated in 7.5% (v/v) n-propanol which suggests an important role of hydrophobic bonds in the association of the gamma-fragment and beta2M. The beta2M did not bind monoclonal IgG from this patient. Several anti-beta2M antisera tested which had been prepared with free beta2M did not react with the beta2M associated with the gamma-fragment, but reacted with the free beta2M obtained by the dissociation of the complex.


Induction of guinea pig antibody responses in vitro. Guinea pig spleen cells cultured together with peritoneal exudate lymphocytes (PEL) were found to generate large numbers of antibody-forming cells (AFC) in vitro in response to hapten-protein antigens. Neither cell type cultured alone yielded appreciable responses. Strain 13 or F1 (Strain 2 X Strain 13) lymphocytes, but not those from strain 2 animals, are able to respond to the genetically controlled antigen, DNP-guinea pig albumin (DNP-GPA). Antisera directed against responder (strain 13) parent Ia antigens selectively blocked the generation of AFC by F1 (strain 2 X strain 13) spleen-PEL mixtures in response to DNP-GPA. Both allogeneic (strain 2) and syngeneic macrophages functioned equally well in presentation of DNP-GPA to strain 13 lymphocytes.


Immunosuppressive factor(s) specific for L-glutamic acid50-L-tyrosine50 (GT). I. Production, characterization, and lack of H-2 restriction for activity in recipient strain. The random synthetic copolymer of L-glutamic acid50-L-tyrosine50 (GT) fails to elicit a GT-specific antibody response in all inbred strains of mice tested. Preimmunization with GT specifically inhibits a GT-MBSA response in certain H2d,k,s, but not other, H-2a,b,q, nonresponder mice. This unresponsiveness is mediated by GT-specific suppressor T cells. Extracts prepared from lymphoid cells of GT-primed suppressor haplotype mice inhibit the development of primary GT-specific antibody responses to GT-MBSA in normal syngeneic mice. Nonsuppressor haplotype mice do not produce GT-specific suppressor factor. The GT-suppressive extract has affinity for antigen and a m.w. of less than 50,000 daltons, thus, resembling antigen-specific immunosuppressive factors already described. However, the GT-suppressive extract does not appear to have H-2 restrictions since it works across allogeneic barriers. Evidence is presented that two genes are required for factor-mediated suppression.


Suppressor cells in tolerance to HGG: kinetics and cross-suppression in high dose tolerance--absence in low dose tolerance. Spleen cells from mice made tolerant with high doses of human gamma-globulin (HGG) specifically suppress the immune response of normal, syngeneic, spleen cells. These suppressor cells were found to be cross-reactive in that they would suppress the immune response of normal spleen cells to bovine gamma-globulin (BGG) as well as to HGG. In contrast, suppressor cells could not be demonstrated in spleens of mice made tolerant with low doses of HGG (i.e., T-cell tolerance), nor could they be found in high dose tolerant mice following a second injection of DHGG at a time when the initial suppressor activity had waned. The role of suppressor cells in the induction, maintenance, and loss of tolerance is discussed.


Multiple polysaccharide antigens of group B streptococcus, type Ia: emphasis on a sialic acid type-specific polysaccharide. Group B streptococci, type Ia (strain 090/14/4), were subjected to sequential extraction procedures and the various extracts were fractionated by a combination of DEAE-cellulose chromatography and gel-filtration. Three major antigens were isolated: the conventional group-specific and type-specific carbohydrates and an acidic polysaccharide consisting of galactose, glucose, glucosamine, and sialic acid. Immunochemical data suggest that the acidic antigen, with the exception of the immunodominant sialic acid, is structurally similar to the conventional pH 2.0 extracted type-specific antigen. Removal of the terminal sialic acid residues from the acidic antigen by mild acid hydrolysis resulted in a residual polymer which was chemically and immunologically analogous to the type-specific carbohydrate. Precipitin analysis focused attention to the cross-reactivity between thia acidic antigen and anti-types Ib, Ic, and III sera.


Antigenic studies of a VH fragment: demonstration of three sets of antigens, idiotypic, VH subgroup, and VH framework-specific antigens. The antigenic properties of the VH region of immunoglobulin heavy chains were studied by means of a fragment corresponding to the variable part of the heavy chain of an IgG3 myeloma protein (KUP) and an antiserum made against this fragment. By hemagglutination, hemagglutination inhibition, and immunofluorescence techniques, it was shown that the anti-VH antiserum detected three sets of antigens in the VH region, namely idiotypic antigens, VH subgroup-specific antigens, and VH domain-(framework) specific antigens. The VH fragment inhibited in a VHII subgroup-specific hemagglutination inhibition test system. The VH fragment was thus antigenically similar to the tvh region found in the intact molecules and the light chains were not needed to express the VH subgroup antigens or the VH framework antigens.


Similar cell surface antigens on hamster cells transformed by different papovaviruses. Transformed baby hamster kidney (BHK) cells were tested for surface antigens by an immunocytoadhesion method. The cells were sensitized with rabbit antisera to cell clones transformed by polyoma or by BK virus and then rosetted with erythrocytes coated with antibody to rabbit immunoglobulin. These antisera detected common antigens on BHK cells transformed by either of three papovaviruses, polyoma, BK, or SV40, but apparently not on normal BHK cells.


In vitro induction of cell-mediated immunity to murine leukemia cells. II. cytotoxic activity in vitro and tumor-neutralizing capacity in vivo of anti-leukemia cytotoxic lymphocytes generated in macrocultures. The main findings of the present study are: (a) highly reactive cytotoxic lymphocytes (CTL) against syngeneic and allogeneic murine leukemia cells were generated in vitro in macro 'one-way' mixed leukocyte-tumor cultures (MLTC). Cultures set up in large tissue culture flasks contained up to 400 X 10(6) normal spleen cells (responder cells) and 20--40 X (10(6) mitomycin C-treated leukemia cells (stimulator cells). Successful sensitization in macrocultures was greatly dependent upon the responder cell density and the responder/stimulator cell ratio. Cytotoxic activity, as measured by the 51Cr-release assay, peaked on day 5--7. (b) Sensitized 'memory' lymphocytes produced in primary MLTC could be restimulated with the original tumor cells to give a more rapid and stronger secondary cytotoxic response. (c) lymphocytes sensitized to allogeneic leukemia cells reacted equally well with sensitizing leukemia cells and with the corresponding normal lymphoid target cells, whereas lymphocytes sensitized to syngeneic leukemia cells did not react with the homologous normal lymphocytes. (d) Cryopreserved normal splenocytes and leukemia cells were as efficient as fresh cells in generating allogeneic and syngeneic CTL. (e) Using a Winn-type tumor neutralization assay, it was shown that both allogeneic and syngeneic splenocytes sensitized in vitro to EL4 leukemia (of C57BL/6 mice) and to YAC leukemia (of A mice) were capable of preventing tumor growth in the syngeneic host, whereas cultured normal splenocytes frequently showed a tumor-enhancing effect. Long-term survivors, remaining after inoculation of leukemia cells and sensitized lymphocytes, also became resistant to a tumor challenge that was up to 10,000 greater than the minimum lethal dose.


Detection by immunofluorescence of common antigenic determinants in unrelated gram-negative bacteria and their lipopolysaccharides. Various gram-negative bacteria were subjected to mild acid hydrolysis. The acid-treated bacteria exhibited strong cross-reactivity with fluorescein isothiocyanate-conjugated antiserum to the Re mutant of Salmonella minnesota. Hydrolyzed bacteria showed considerably stronger fluorescence than heat-treated bacteria. It is assumed that acid hydrolysis uncovers shared glycolipid determinants that are responsible for cross-reactivity. Isolated homologous and heterologous lipopolysaccharides were allowed to react with antibody to S. minnesota Re insolublized by covalent binding to aminohexyl Sepharose 4B. The resulting antigen-antibody complexes were visualized by exposure to the fluorescent antiserum. This treatment allows the demonstration of glycolipid structures of intact lipopolysaccharides.


Correlation of e antigen, DNA polymerase activity, and Dane particles in chronic benign and chronic active type B hepatitis infections. The e determinant of hepatitis B surface antigen (HBS Ag) was found in 23 of 42 patients with chronic hepatitis B virus (HBV) infection. Presence of e antigen was associated with increases in DNA polymerase activity and in the number of circulating Dane particles. In the group with detectable e antigen, the average DNA polymerase activity was 367+/-78 counts per minute (cpm; mean+/-standard error [SE]), and the average number of Dane particles counted in electron micrographs was 4.4% of the total HBS Ag. In contrast, e antigen-negative patients had an average DNA polymerase activity of 40+/-6.9 cpm (P less than 0.1) and an average Dane particle count equal to 0.6% of the HBS Ag. The e antigen was detected in 68% of patients who were HBS Ag carriers or had persistent viral hepatitis and 40% of those with chronic active type B hepatitis. Thus, the presence of e antigen correlated with both the chronicity and presence of infectious HBV. However, it did not correlate with the type or severity of liver disease after HBV infection, since e antigen was present in both chronic benign and chronic aggressive hepatitis B infections.


Immunologic studies in pneumococcal disease. Many patients die from pneumococcal disease despite the availability of effective antimicrobial agents. Immunologic studies including detection, typing, and quantitation of serum pneumococcal capsular polysaccharide (PCP) antigen by counterimmunoelectrophoresis (CIE), quantitation of PCP antibody by radioimmunoassay (RIA), and quantitation of serum complement components C3, C4, and C3PA and serum immunoglobulins IgG, IgM, and IgA by the radial immunodiffusion technique of Mancini were performed with the sera of 18 patients. Five patients died (group I), and 13 survived (group II) pneumococcal infection. Both groups were comparable in age, underlying disease, and leukopenia on admission. All patients of group I and 10 of 13 (77%) of group II patients were bacteremic. Two patients in each group had an extrapulmonary focus infection. PCP antigen was detected in the sera of all group I and nine of 13 group II patients. PCP antigen levels were larger than or equal to 15 microng/ml in four of five group I and two of 13 group II patients (p = 0.022). Levels of antibody to PCP exceeded 100 ng/ml of antibody nitrogen (AbN) in 10 of 12 group II and one of five group I patients (p = 0.027) during the course of illness. All group I patients and three of 12 group II patients had decreased levels of one or more complement components on admission (p less than 0.01). One or more complement components remained decreased until death in four group I patients but returned to normal or elevated levels in all group II patients. No difference in serum immunoglobulin concentrations were found.


Catabolism of rat beta2-microglobulin in the rat. Highly purified rat beta2-microglobulin (beta2m) as well as cytochrome c and lysozyme were radiolabeled and their catabolism studied in the rat. More than 90 percent of these low molecular weight proteins were removed from the serum within an hour and excreted into the urine by 24 hours. Except for the kidney in which the concentration of these protein is ten- to twentyfold greater than in the serum, there is little evidence that rat tissues are concentrating these proteins. The stomach was found to concentrate radioiodine. The catabolism of rat beta2m differed from that of cytochrome c and lysozyme in that the kidney contained twice as much labeled rat beta2m. In addition, the rat excretes 10 to 15 percent of the injected rat beta2m but only 1 to 5 percent of the cytochrome c or lysozyme. These studies established a basis for turnover studies of beta2m complexed with other cell membrane proteins, for example, HL-A or H-2 peptides.


Two-site assay of bovine parathyroid hormone. A two-site assay has been developed for bovine PTH. This technique involves reaction of the antigen with two antibody molecules with the purpose of increasing specificity. In paratice the hormone was extracted form plasma samples on to plastic tubes coated with antibody specific for PTH (1-34). Uptake was then measured using 125I-labelled antibodies specific for PTH (53-84). In this way a sensitive assay was obtained for PTH (1-84) which did not recognize molecular fragments. This technique was used in conjunction with immunoradiometric assays specific for either NH2-terminal (1-34) or CO2H-terminal (53-84) molecular fragments to study the clearance of infused PTH in the cow. Preliminary results support the hypothesis that the intact molecule is rapidly degraded in the peripheral circulation with the preferential disappearance of an NH2-terminal fragment. Studies on endogenous secretion during calcium and EDTA infusions indicated that there was little intact hormone present at the times when CO2H-terminal immunoreactivity was readily measured.


Tissue culture studies in Hodgkin's disease: Morphologic, cytogenetic, cell surface, and enzymatic properties of cultures derived from splenic tumors. Monolayer and suspension cell cultures prepared from Hodgkin's disease tumors in the spleen were examined microscopically and by cytogenetics, tested for lymphocyte and monocyte cell surface properties, and assayed for enzymes by histochemical and spectrophotometric techniques. Hodgkin's disease monolayer cultures were composed of rapidly proliferating round and polygonal cells that were capable of propagation in vitro for an indefinite period of time. Abnormal aneuploid chromosomes were found in short-term Hodgkin's disease monolayers that had been passaged 16-20 times, and in established cell lines carried in culture longer than 3 yr and passaged more than 200 times. Cells fromHodgkin's disease monolayers contained lysozyme (muramidase), fluoride-resistant alpha naphthol acetate esterase, acid and alkaline phosphatase, and chymotrypsin-like activity. The monolayers did not exhibit specific cell surface markers or phagocytosis. Suspension cultures derived from Hodgkin's disease monolayers were composed of cells with aneuploid karyotypes and similar enzymes. The Hodgkin's disease suspension culture cells had surface receptors for complement and IgGFc, lacked surface or cytoplasmic immunoglobulin, and did not form Erosettes, react with an antithymocyte serum, nor exhibit phagocytosis. Normal monolayer culture cells, derived from adult spleen and human fetal spleen and thymus, were composed of spindle cells with a diploid number of chromosomes that could be carried for only a finite period of time in vitro. Normal cultured cells contained similar esterases and phosphatases, but were devoid of lysozyme and chymotrypsin-like activity. The morphologic, cytogenetic, cell surface, and enzymatic findings indicate that our Hodgkin's disease monolayer and suspension cultures are composed of cells with many properties suggesting an origin from monocytes (macrophages) rather than lymphocytes or fibroblasts. The presence of aneuploid karyotypes is consistent with a neoplastic origin and derivation from a malignant cell of Hodgkin's disease.


Mutation in a new H-2-associated histocompatibility gene closely linked to H-2D. Sequential precipitations of soluble BALB/c antigen with antisera detecting private and public H-2 specificities indicated three distinct classes of molecules of 45,000 mol wt. However, only two of these classes of molecules were detectable in antigen from the loss mutant, BALB/c-H-2db. The class of molecules, detectable in the wild-type strain but missing in the mutant, does not bear private specificities but does react with an antiserum detecting H-2 public specificities. Absorption in mutant mice of the antiserum to public specificities, left antibodies specific for the antigen detectable in BALB/c but not BALB/c-H-2db. Genetic mapping studies using this specific antiserum indicated that the antigenic loss of this mutant is in a gene which maps in or close to the H-2D region, separable from the H-2K, S, G, Qa-2, and Tla regions.


Cultured human monocytes synthesize and secrete alpha2-macroglobulin. alpha2-Macroglobulin levels in the supernates of cultures of different subpopulations of human peripheral blood mononuclear leukocytes were assayed by a radioimmunoassay. Unfractionated mononuclear leukocytes produced greater amounts of the macroglobulin (4.0 vs. 0.8 ng/10(6) cells) than did subpopulations enriched in T or B+T lymphocytes, by passage through nylon wool or cotton wool columns, respectively. Still higher concentrations of alpha2-macroglobulin (40 ng/10(6) cells) were measured in the supernates of glass-adherent mononuclear leukocyte cultures. These results suggest that cells of monocyte-macrophage lineage are mainly, if not exclusively, responsible for the appearance of alpha2- macroglobulin in the supernate of human peripheral blood leukocyte cultures. The de novo synthesis and release of alpha2-macroglobulin by cultured monocytes was demonstrated by immunoprecipitation of radioactivity from supernates of 32S-methionine-labeled glass-adherent cells. Antiserum against purified alpha2-macroglobulin was used in both Ouchterlony double diffusion and double antibody precipitation tests. SDS-polyacrylamide gel electrophoresis of immunoprecipitates showed that most of the radioactivity comigrated with authentic alpha2-macroglobulin subunit at about 160,000 daltons.


Evaluation of audiovisual teaching materials in family practice: a report of review activities 1974-1976. Recent years have seen increasing interest in the use of audiovisual teaching materials throughout medical education. There is good evidence that different individuals learn best in different ways according to their particular learning styles, personalities, and learning needs. Audiovisual materials can provide flexible approaches to meet varied learning needs and facilitate self-instruction. In the past, there has been no systematic method of identification and evaluation of audiovisual teaching materials relevant to family practice. This paper describes the experience of a review process which was initiated three years ago. A complete list is presented of audiovisual teaching materials which have been reviewed for family practice and future directions of the review process are discussed.


Polyneuropathy in hypothyroidism. Clinical and nerve biopsy study of 4 cases. Clinical and sural nerve biopsy findings are described in four hypothyroid patients with manifestations of a diffuse peripheral neuropathy. Clinical examination revealed distal sensory inpairment, complaints of burning  and lancinating extremity pains, ataxia and a decrease of deep tendon reflexes with total ankle jerk loss. In one patient, signs of the peripheral neuropathy comprised the leading symptomatology of the underlying metabolic disorder. Light and electron microscopic investigations of the sural nerve biopsies revealed a marked reduction of myelinated fibers, affecting mainly the large myelinated axons. The histograms showed a pathological unimodal spectrum of myelinated fiber diameters. Dystrophic and degenerative changes in axons of all fiber classes and the presence of clusters of thinly myelinated small fibers suggest that neuroaxonal degeneration and incomplete regeneration may represent the major pathogenetic mechanism of the neuropathy in these hypothyroid patients.


[The frequency of rapid eye movements and blinks as an activation indicator (author's transl)]. Electrooculographic (EOG) records were studied in 10 neurological patients with disturbances of activation according to clinical assessment and in 12 normal subjects. Rapid horizontal eye movements (saccades) and movements of the eyelids (blinks) were measured. It was concluded that the frequency distribution of blinks and saccades indicates activation deficits in a sensitive and reliable way. The continuous decrease of activation from orthological to pathological states could be shown by counting the frequencies of blinks and saccades. The relationship between the two variables is interpreted with regard to its functional significance.


A psychometric evaluation of the acute tremulous state. The acute alcohol withdrawal state (tremulous state), with mainly vegetative symptoms and without evident loss of conciousness or confusion, was evaluated as to functional psychopathological disturbances aiming to present a complete and objective record of the clinical findings and to establish a control of the course and drug treatment. We tried to meet the inherent inability to cooperate by using proven and also new test devices (flicker fusion, simple reaction time on light and tone, reaction on multiple serial stimuli, visual motor coordination, tachistoscopic perception and memory test, test of concentration and sustained performance with simple arithmetical calculation by analogy with the Pauli test) to circumvent the difficulties arising when patients have to answer long questionnaires. The tests enabled a measurement of the disturbances as objective as possible and proved to have a discriminating sensitivity for different functions. The correlations between the results were found to be similar for alcoholics and controls. tthe degree of the established functional cerebral and cerebellar defects which was revealed was more severe than expected in this mild stage of withdrawal.


Electrolyte changes and acid base balance after alcohol withdrawal, with special reference to rum fits and magnesium depletion. 70 chronic alcoholics in the withdrawal state, 45 with convulsions and 25 controls without convulsive seizures, were tested with respect to electrolyte changes and acid base balance in serum or blood and cerebrospinal fluid (CSF). It was of special interest to note that there was a partial independence between magnesium levels in serum and CSF. Thus the serum level has only a limited liability as to magnesium depletion suggested to be responsible for seizure precipitation. In the seizure group a slightly but significantly lower magnesium, potassium and calcium in CSF and a significant decrease of potassium and calcium in serum were revealed. In the nonzeizure controls a similar decrease of magnesium in serum and potassium in CSF was observed while serum potassium and calcium in CSF and serum remained in low normal range. In both groups there was a prominent respiratory alkalosis. The role of magnesium depression for seizure precipitation is discussed with respect to the concomitant changes of other electrolytes and acid base disturbances.


[The pathogenetic importance of the affection and of the therapy of MS-patients' teeth in comparison to those of epileptics (author's transl)]. 51 MS-patients between 18 and 50 years and an equally old control group of 51 epileptics were dentally and roentgenologically investigated. Especially the fillings of amalgam were observed. This quantitative registration showed a prevailing of 18.5% among the epileptics without statistical significance. On the other hand MS patients suffered more frequently from carious teeth. Both groups are extremely unsufficiently provided with prosthesis. 85% of all MS-patients and 71.4% of all epileptics showed progressive paradontopathics. Only 4 of 40 epileptics treated with hydantoins produced a hyperplasia of their gingivas. In view of odontogenic foci the clinical and the roentgenological results could not reveal any differences between the both groups of patients. Only granulomas of the teeth were more frequent among the MS-patients. Baasch's theory of the amalgam-etiology of MS is discussed and rejected because of lack of evidence. The problem of the focus therory is generally and specially discussed, too, as a possibility causing MS. But a connection between focus and MS is not sure in spite of striking investigations. As a general result conclusions should be drawn from the bad conditions of the MS patients' teeth with regard to a better dental providing.


Regeneration of cauda equina fibres after transsection and end-to-end suture. Light and electron microscopic study in the pig. The regeneration of nerve fibres in pigs was studied three months after transection and end-to-end suture of the dorsal and ventral spinal nerve roots L5 and S1. Although no detailed quantitative analysis was performed, it was clearly evident that regeneration in motor roots is significantly more efficacious than in sensory roots. Probably as a reaction to Wallerian  degeneration, an increase of histiocytes, fibroblasts and collagen fibrils, as well as a thickening of the sheaths was observed in the transected and sutured roots. Heterotopic glial bundles were found in the regenerated dorsal roots.


Ultrastructural study of a muscle biopsy from a patient with subacute myelo-optic neuropathy. A man, aged 46, who had been taking Clioquinol in high doses for a long period, developed a characteristic neurological syndrome of subacute myelo-optic neuropathy rather abruptly. Electron microscopical examination of the muscle biopsy, obtained five months after the onset of the disease, revealed severe degenerative changes of the presynaptic nerve endings and some unique paracrystalline inclusions in the sole plate region. The latter may represent the morphological expression of the toxic agent which is held responsible for the subacute myelo-optic neuropathy.


Anterior commissurotomy for intractable pain. Anterior commissurotomy for the relief of intractable lower half and midline pain has been found to be a very satisfactory procedure in a small group of severely incapacitated patients. The presumed anatomic rationale for undertaking the procedure has not been confirmed by postoperative sensory examinations. Alternative explanations for its effectiveness remain conjectural. The operative procedure is described.


Autoimmune disease antigens.

Autoimmune disease antigens. The first step towards understanding the cellular interaction which results in autoimmune disease is to determine what triggers the recognition between a specific autoimmune antigen determinant and the cellular receptor. In this review, we have focused on the antigen inducing experimental allergic encephalitis (EAE) because the antigen has been characterized and a relatively large body of information on its biological activities has been accumulated. Clearly, a specific allergic encephalitis-producing determinant is present and is represented on a relatively small portion of the molecule. The determinant induces a wide variety of biological reactivities, some of which are classed as cellular mediated. An attempt is made to dissect activities such as blast transformation (BT), migration inhibitory factor (MIF), in vivo delayed type hypersensitivity reaction (DTH) and EAE and to relate them to the structural requirements which the determinants possess. The complexities which arise indicate that subpopulations of cells with different receptor activities may respond selectively and that recognition of the receptor is produced by an EAE determinant consisting of three amino acids in a specific linear sequence. Furthermore, under experimental circumstances the EAE activity can be dissociated from the other activities (BT, MIF, DTH), indicating that while these tests are used generally to follow various human autoimmune disease activities, they may represent the reaction of a broad spectrum of cells.


Changes in serum and plasma proteins and in IgG and IgM antibodies in calves experimentally infected with sarcocystis from dogs. Fifteen calves were used in three experiments to determine changes in serum and plasma proteins and IgG and IgM levels after oral inoculation with Sarcocystis sporocysts from dogs. Total serum or plasma protein levels in inoculated calves decreased during the acute phase of infection (4 to 5 weeks after inoculation) and then increased and became greater than the levels of control calves 7 to 8 weeks after inoculation. The initial decrease in total protein reflected reduced serum albumin and the subsequent increase reflected increased immunoglobulin levels. Immunoglobulin levels increased in both IgM and IgG fractions. Specific antibody activity against Sarcocystis antigen was 6 to 9 times greater in the IgM than in the IgC fraction 5 weeks after inoculation, but IgG activity became approximately 17 to 27 times greater than IgM activity 10 to 13 weeks after inoculation.


Further observations on the specificity of antigen 5 of Echinococcus granulosus. The presence of IgE antibodies to antigen 5 of Echinococcus granulosus was detected by means of radioimmunoelectrophoresis in the sera of two of six patients infected with E. multilocularis. Sera from three of these patients gave a precipitin band in gel diffusion tests identical to that produced by a monospecific rabbit anti-E. granulosus antigen 5 serum, when tested against whole hydatid fluid. Sera from 19 individuals infected with Fasciola hepatica, 20 with Schistosoma mansoni, and 5 with with Taenia saginata showed no detectable antibodies against antigen 5 of E. granulosus, The monospecific rabbit anti-E. granulosus antigen 5 serum did not react in immunodiffusion with homologous antigen when absorbed with either 4 mg/ml of whole hydatid fluid or with 200 mg/ml of a soluble E. multilocularis extract. Absorption of the monospecific antiserum with crude antigens of either F. hepatica, Onchocerca volvulus, S. mansoni, or T. saginata did not abolish the reaction with antigen 5. It appears, therefore, that antigen 5 can no longer be considered specific for E. granulosus, but is also present in E. multilocularis. In the light of this observation, some reevaluation of immunodiagnostic tests in hydatid disease will be necessary.


Keys to the genera of chiggers of the western hemisphere (acarina: trombiculidae). Synotpic keys to the 87 genera of chiggers in the Western Hemisphere (Nearctic and Neotropical regions) as well as illustrations to the terminology employed, are presented.


Surface characterization of temporary restorations: guidelines for quality ceramics. A technique was presented that enables a dentist to characterize a temporary restoration and its duplicate with external surface stains. This adjusted duplicate and an accompanying zone-diagrammed laboratory work authorization order will provide the dental laboratory technician with accurate information for making a porcelain restoration with esthetic acceptability.


Tumor-associated antigen levels (carcinoembryonic antigen, human chorionic gonadotropin, and alpha-fetoprotein) antedating the diagnosis of cancer in the Framingham study. Determinations of carcinoembryonic antigen (CEA), human chorionic gonadotropin (HCG), and alpha-fetoprotein (AFP) were done by use of frozen serum samples antedating the diagnosis of cancer for 9 pancreatic and 8 gastric carcinoma patients from the Framingham Heart Study. The longest intervals for elevated antigens before cancer diagnosis were 10 months for CEA and 26 months for HCG. (The single elevated AFP was found in a sample 10 days before clinical diagnosis.) Samples from 31 controls matched with the cancer subjects by age, sex, vital capacity, and smoking status showed over 20% "false" positive CEA elevations (all smokers with low vital capacities) and over 20% borderline false positive HCG elevations in postmenopausal females. Although 10-26 months' lead time could infer some potential for use of these tumor-associated antigens to help detect malignant neoplasms at an earlier stage, a serious problem of frequent false positives prevents CEA and HCG levels from being useful as cancer-screening tests at this time.


Forecasting the development of primary hepatocellular carcinoma by the use of risk factors: studies in West Africa. An association between hepatitis B virus (HBV) and primary hepatocellular carcinoma (PHC) has been found in several studies in Africa, Asia, and elsewhere. In this paper we considered the interrelations between several events related to HBV infection, which include the presence of: 1) hepatitis B surface antigen (HBsAg), 2) antibody to hepatitis B core antigen (anti-HBc), 3) antibody to the surface antigen (anti-HBs), 4) chronic liver disease, 5) elevated alpha-fetoprotein, and 6) PHC. With the use of preliminary epidemiologic data, risk factors related to these events were calculated. We suggested that the interactions between these events and HBV infection in parents be used to estimate the risk of PHC for an individual in this environment.


Cell-mediated cytotoxic response to cells bearing Marek's disease tumor-associated surface antigen in chickens infected with Marek's disease virus. In a microcytotoxicity test in which 51Cr-labeled cells of a Marek's disease (MD) lymphoblastoid line (MSB-1 line) were used, cell-mediated cytotoxicity of spleen cell suspensions prepared from chickens inoculated with MD virus was demonstrated. This cytotoxic response, presumably directed against MD tumor-associated surface antigen, was detectable briefly after virus infection and paralleled the appearance of early lymphoproliferative lesions characteristic of MD.


Testing of pesticide toxicity in tissue culture. The effect of pesticides on cell growth and on the cell layer was examined. The pesticides under study had a similar toxicity for the cell suspension and for the cell layer. Vital staining of the cells allowed a more sensitive determination of the cytotoxic concentrations than did the studies on cell morphology without staining. No correlation was found between the cytotoxic concentrations and LD50 values of mammals. The results were compared with those of earlier experiments on rats in which methods of general toxicology and neurotoxicology were applied. Of the eight compounds tested, in six the tissue culture method proved even more sensitive than did the functional neurotoxicological tests. It is suggested that the in vitro method with cultured cells should be used to complement in vivo functional toxicological procedures.


Characterization of an RNA-directed DNA polymerase found in association with murine intracytoplasmic A-particles. An RNA-directed DNA polymerase was found to be associated with intracytoplasmic A-particles from DBA/2 mouse leukemia cells. The enzyme activity was detected after disrupting the purified particles with 2 M NaCl-20 mM dithiothreitol. The presence of a divalent cation and all four deoxyribonucleoside triphosphates was essential for this enzyme activity. The enzyme had a clear preference for Mg2+ over Mn2+. Cesium sulfate isopycnic gradient centrifugation of the DNA product synthesized in the actinomycin D-containing reaction revealed the presence of DNA-RNA hybrid. Furthermore, the purified DNA product was found to hybridize with RNA isolated from A-particles. These observations strongly indicate that the endogenous A-particle RNA serves as the template for the DNA polymerase.


Assay of noninfectious fragments of DNA of avian leukosis virus-infected cells by marker rescue. A marker rescue assay of noninfectious fragments of avian leukosis virus DNAs is describe. DNA fragments were prepared either by sonication of EcoRI-digestion of DNAs of chicken cells infected with wild-type Rous sarcoma virus, with a nontransforming avian leukosis virus, and with a mutant of Rous sarcoma virus temperature sensitive for transformation. Recipient cultures of chicken embryo fibroblasts were treated with noninfectious DNA fragments and infected with temperature-sensitive mutants of Rous sarcoma virus defective in DNA polymerase or in an internal virion structural protein. Wild-type progeny viruses which replicated at the nonpermissive temperature were isolated. Some of the wild-type progeny acquired both the wild-type DNA polymerase and the subgroup specificity of the Rous sarcona virus strain used for preparation of sonicated or EcoRI-digested DNA fragments. Therefore the genetic markers for DNA polymerase and envelope were linked and appeared to be located on the same EcoRi fragment of the DNA of Rous sarcoma virus-infected cells.


Radioimmunoassay for mammalian type C viral reverse transcriptase. Radioimmunological techniques were applied to the analysis of reverse transcriptase of mammalian type C RNA viruses. The polymerase of Rauscher mouse leukemia virus was purified by ion exchange and sequential affinity chromatography. Radioimmunoassays that utilized the viral enzyme as a probe detected as little as 1 ng of purified polymerase. No cross-reactivity could be demonstrated between the reverse transcriptase and other known virus-coded proteins. By comparing the immunological reactivity of the purified enzyme with the reactivity of detergent-disrupted virions, Rauscher mouse leukemia virus was shown to contain the antigenic equivalent of 40 molecules of reverse transcriptase. In a homologous competition immunoassay, the Rauscher viral enzyme demonstrated type-specific antigenic determinants, which distinguish it from other mouse type C viral polymerases. In a broadly reactive interspecies immunoassay, the reverse transcriptases of a number of mammalian type C viruses were cross-reactive, indicating their shared antigenic determinants. Various treatments that inhibit or inactivated DNA polymerase activity had little or no effect on the immunological properties of the enzyme. Thus, radioimmunoassays should be useful in the search for type C viral reverse transcriptase as a marker of subviral expression.