Analysis of some histochemical colour reactions for cytospectrophotometry of basic and acidic proteins in nervous tissue cells. Cytospectrophotometric analysis of model systems and of single cells in Carnoy-fixed paraffin sections of the nervous tissue has shown two histochemical colour reactions for basic proteins and two reactions for acidic proteins to obey Bouger-Lambert-Beer's law. Basic proteins can be stoichiometrically stained either with Fast Green FCF, pH 8.2, or with heparin-Alcian Blue; acidic proteins either with Fast Green, FCF, pH 2.6, or with Toluidine Blue o. basing on better reproducibility of cytochemical results and sufficiently good preservation of cell structure, heparine-Alcian Blue is recommended for cytospectrophotometric determinations of basic proteins. To stain acidic proteins both Fast Green, pH 2.6, and Toluidine Blue O are recommended, each of the dyes being characterized by its own advantages and disadvantages.


Improvement of the micromethod for the limulus lysate test. Frauch's micro-slide method was improved to facilitate the endpoint-determination of the Limulus test. Two precise observations, by inverted phase contrast microscopy and with a staining procedure, were newly performed as additions to the slide test. The staining procedure was proposed as an improved method for the Limulus test since it is simple and convenient. In the staining method, bromophenol blue (BPB) solution was used as the staining solution. A negative (-), a strong positive (++) and a weak positive reaction (+) were characterized by a "ring" formation, a "cloud-like" spread of gel and a "spot" in the "cloud" respectively. Since the distinction between (-) and (+) reactions was obvious in the proposed method, determination of the endpoint was easier than in the ordinary tube and Frauch's method. The sensitivity of the present method was equal to or higher than that of other methods. Inverted phase contrast microscopy was utilized to confirm the findings obtained by the staining method. The volume of the lysate used in this method was as little as 1/10 of that used in the tube method.


[Lithium-intoxication in a newborn (author's transl)]. A newborn, whose mother had been treated with lithium carbonate for a manic-depressive illness suffered from hypotonia, poor feeding, weak Moro reflex, hyperexcitability, thyroid enlargement and cyanosis. Symptoms resolved within a few weeks. No malformations could be detected. The psychomotor development of the child was normal up to the age of 18 months.


Theoretical mechanisms for synthesis of carcinogen-induced embryonic proteins. II. Perturbations of post-translational nuclear protein modification. An hypothesis of carcinogenesis is derived which suggests that carcinogens initially induce derepressions of structional genes of post-translational modifying enzymes. Although the resulting cascade of changes in genic activity would be measureable because of the large number of differentiated cells present in a tissue, only stem cells are vulnerable to the neoplastic array of genic expression by virtue of their repressors being more vulnerable to conformational modification. Furthermore it is derived that there are at least three major classes of repressors and the repressors that are synthesized during embryogenesis are particularly susceptible to derepression mechanisms.


Genetic activity of bleomycin: differential effects on mitotic recombination and mutations in yeast. The genetic effects of the antitumor antibiotic, bleomycin were studied in different strains of Saccharomyces cerevisiae. It was observed that the drug induced a high frequency of mitotic recombination and gene conversion. In contrast, it produced only a few mutations from adenine independence to adenine dependence and histidine dependence (a missense mutant) to histidine independence. In the strains carrying ochre-suppressible auxotrophic markers, no prototrophs were induced by this drug. The results indicating the specific activity of bleomycin are discussed and, in this connection, the usefulness of yeast as a test organism in mutagenicity screening is emphasized.


Increased transport of (G-14C) cytidine into rat liver after administration of alpha-hexachlorocyclohexane. The transport of cytidine into liver over a wide dose range (0.003--100mumol per animal) proceeds as an nonsaturabel process. After the administration of alpha-hexachlorocyclohexane (alpha-HCH)1 the transport of (G-14C) cytidine is markedly activated if the concentration of nucleoside administered is low; the differences between the control and experimental group disappear after the administration of higher doses of the nucleoside. The incorporation of (G-14C) cytidine into RNA cytosine is enhanced after the administration of alpha-HCH. The degree of utilization of labeled cytidine for the RNA synthesis decreases in proportion to the logarithm of its dose, both in the control and in the experimental group. Increased transport of (G-14C)cytidine is observable even between the 4th and 6th day after the administration of a single dose of alpha-HCH.


Chlordecone intoxication in man. I. Clinical observations. Industrial overexposure to chlordecone, an organochlorine insecticide, caused tremor in 76 of 148 exposed workers. Chlordecone was absorbed through oral, respiratory, and dermal routes, the last possibly the most significant. Epidemiology of this incident disclosed low-level, widespread environmental exposure of man to chlordecone. In 23 workers with chronic chlordecone intoxication, tremor was associated with opsoclonus, pleuritic pain and arthralgia. No seizures were reported. The site of action of chlordecone on the central nervous system is unknown. It concentrates in human adipose and hepatic tissue but is not biodegradable, either in humans or elsewhere in nature.


Chlordecone intoxication in man. II. Ultrastructure of peripheral nerves and skeletal muscle. Tremors, mental changes, opsoclonus, muscle weakness, gait ataxia, incoordination, and slurred speech developed in several employees in a Virginia chemical plant during the summer of 1974. Epidemiologic and clinical studies suggested that the chlorinated insecticide chlordecone (Kepone) was responsible. Severity of symptoms seemed directly related to dose and duration of exposure. Five sural nerve and six muscle biopsy specimens were examined by light microscopy and electronmicroscopy. The sural nerves were also evaluated by computerized morphometry, which showed considerable decrease in the number of unmyelinated fibers and lesser abnormalities of myelinated fibers. Compared with the nerves of the control subjects, those of patients may have had an increase in Reich and Elzholz bodies, and a modest increase in endoneurial collagen. There were occasional "collagen pockets," stacks of Schwann cell cytoplasmic membranes, redundant Schwann cell cytoplasmic folds, and fewer unmyelinated axons. The skeletal muscles contained increased amounts of lipofuscin and lipidlike droplets in subsarcolemmal areas and within intermyofibrillary spaces; the significance of this is unknown. Fiber size variability, type I predominance, and type grouping were present in three cases. All results strongly suggest that chlordecone is a neurotoxic agent predominantly affecting Schwann cells and unmyelinated fibers of peripheral nerves.


Studies of sarcolemmal integrity in myopathic muscle. Studies of sarcolemmal integrity with extracellular Procion Yellow gave evidence of structural defects in a high percentage of muscle fibers in Duchenne dystrophy and in vitamin E-deficiency myopathy in the guinea pig. The dye entered fibers in these conditions more often than in controls. Myofibrillary clumping occurred in the region of major penetration of the dye. It seems likely that sarcolemmal defects occur in vivo in these conditions, allowing the entry of high concentrations of calcium into the sarcoplasm with consequent fiber damage. The biochemical basis of these sarcolemmal defects remains to be elucidated. Our procedure for biopsy of the vastus medialis muscle may prove useful in preparing a limb muscle for in vitro physiologic studies.


[Reversible suppression of synaptic function by anti-synaptosome-antibodies (author's transl)]. We report about experiments to isolate Synaptosome-Membranes of the rat cerebellum in order to produce Antibodies directed toward these membranes in rabbits. The raised Antisera were tested in different Complement-fixation-assays. They reacted only with the Synaptosome-Membranes, not with heart, kidney, liver- and spleen-membranes. Different fractions of the isolation-procedure for synatosomal membranes were also checked for their ability to fix Complement. Only Synaptosomes and the synaptosomal membranes gave sufficient Complement fixation, while Mitochondria, soluble brain proteins, synaptic vesicles and braincell nuclei did not. These biochemical and immunological findings are further supported by electrophysiological data, where the transsynaptic component of the parallel fibre-evoked field potential was strongly suppressed by the administration of the immune sera containing antibodies against membranes of synaptosome fractions.


Pituitary injection of alcohol for widespread cancer pain. Moricca's technique of the pituitary injection of alcohol has been performed 155 times on 92 patients at the Centre for Pain Relief Walton Hospital, Liverpool. There were 4 post-operative death within the first week. The technique is considered to be an exceedingly useful method of relieving pain in patients suffering from widespread inoperable cancer. 40% of the patients treated had complete relief of their pain, half of these (20%) having complete relief for at  least 4 months. 30% of the patients treated had some relief of pain and 30% had none. During injection, it was noted that Myodil injected before the alcohol spread along the  pituitary stalk and the walls of the third ventricle. It eventually broke through into the third ventricle. It is postulated that the alcohol follows this route and the relief of pain is due to destruction of thalamic and hypothalamic nerve pathways.


Scabies in children.

Scabies in children. We are in the midst of a pandemic of scabies. Classic scabies is seen less frequently in the current cycle. The infestation frequently occurs in special forms, the differential diagnosis of which includes nearly every pruritic dermatosis. The cause of the current pandemic is not clear but there is evidence that immunologic factors, particularly delayed hypersensitivity, play a principal role. The diagnosis should be made with certainty, preferably by identifying the mite prior to instituting therapy. The modern scabicides are highly effective. A recent re-evaluation of the risk-benefit relationships of lindane has led to altered therapeutic routines.


Alpha2-macroglobulin deficiency in a patient with Ehlers-Danlos syndrome. A new genetic defect, alpha2-macroglobulin deficiency, was found in a patient with Ehlers-Danlos syndrome (EDS). Other members of the family of five exhibiting this abnormality were the mother and one sister. All members, including the patient, had normal serum albumin and alpha1-antitrypsin levels. The deficiency, reported here for the first time, appears to be inherited by an autosomal co-dominant mode. Statistical evaluation of the dihybrid crosses for independent assortment between EDS and hypo-alpha2-macroglobulinemia showed a probability of 0.7 to 0.75. However, a possible link between EDS and hypo-alpha2-macroglobulinemia is suggested since the observed ratios of four siblings are exactly as expected, assuming that double gene defects are linked in the mother's genotype.


Reverse transcriptase: a monitor for perturbation effects of spin labels covalently bound to nucleic acids. The sensitive biological assay for reverse transcriptase was used to monitor potential perturbation effects of spin labels covalently bound to various nucleic acids or nucleic acid analogs to the extent of about one label per 100 residues. The inhibitory properties of the spin labeled and unlabeled biopolymers were compared for evaluating possible interference of the reporter group in protein-nucleic acid interaction studies. The amount of inhibitor required for 50% inhibition (ED50) was determined for the competitive inhibitors (U)n, l(U)n, (RUGT,U)n, (Um)n, (A)n, (Am)n, and l(A)n as well as for thenon- or uncompetitive inhibitors (dUfl)n, l(dUfl)n, (dUz)n, and l(dUz)n. The most pronounced inhibition was observed with spin labeled and unlabeled (dUfl)n. The results indicate that the ED50 and the kinetic patterns of inhibition are similar for the spin labeled and unlabeled inhibitors studied. Thus, the presence of a limited number of spin labels in a nucleic acid matrix has little effect, if any, on reverse transcriptase-nucleic acid complexes and most likely on other protein-nucleic acid complexes.


Somatic cell hybrids producing antibodies specific for the tumor antigen of simian virus 40. We have produced somatic cell hybrids between mouse myeloma cells deficient in hypoxanthine phosphoribosyltransferase IMP: pyrophosphate phosphoribosyltransferase; EC 2.4.2.8) and spleen cells derived from mice primed with either syngeneic or allogeneic cells transformed by simian virus 40. Such hybrids produced antibodies specific for simian virus 40 tumor (T) antigen. Only four of twelve independent hybrid cell cultures produced antibodies against simian virus 40 T antigen that crossreacted with the T antigen induced by BK virus, a human papovavirus isolated from patients who had undergone immunosuppressive therapy.


Role of Na+ in alpha-aminoisobutyric acid uptake by membrane vesicles from mouse fibroblasts transformed by simian virus 40. The uptake of alpha-amino[(3)H]isobutyric acid (AIB) was studied in membrane vesicles from mouse fibroblasts transformed by simian virus 40 to examine the features of the Na(+)-stimulated and Na(+)-dependent AIB transport process. The simultaneous addition of NaCl and AIB to these vesicles produced a transient accumulation, or "overshoot," of amino acid 3-4 times the equilibrium value. Both the initial rate of uptake and the rate of fall of intravesicular AIB after maximal accumulation were sensitive to the temperature of incubation. The overshoot of AIB uptake was enhanced with Na(+) salts of highly permeant lipophilic anions, such as SCN(-) and NO(3) (-), and was decreased by the addition of SO(4) (2-), a relatively impermeant ion. Gramicidin D, which enhances the membrane conductance of Na(+) electrogenically, decreased the overshoot, while a potassium diffusion potential, induced by valinomycin (in K(+)-preloaded membrane vesicles), produced a Na(+)-dependent overshoot of AIB uptake. When vesicles were preincubated with both Na(+) and AIB, followed by the generation of an interior negative membrane potential (by the addition of SCN(-)), an overshoot of AIB uptake resulted. However, this did not occur in the absence of Na(+). It is concluded that, apart from its role in the generation of a transmembrane electrochemical potential, Na(+) is essential for the overshoot of AIB uptake.


Mechanism of antigen-induced antibody biosynthesis from antibody precursors, the heavy and light immunoglobulin chains. The immediate precursors of antibody molecules, the heavy (H) and light (L) peptide chains of the immunoglobulins, combine with each other by means of disulfide bonds formed by dehydrogenation of their cysteine residues. In the absence of an antigen this process yields the heterogeneous mixture of normal immunoglobulins. Antigens or their processed derivatives (Ag) interfere with this stochastic process by noncovalent combination with complementarily fitting H chains. The (Ag.H)(n) complexes thus formed, owing to the loss of rotational and translational freedom, combine preferentially with those L chains whose V(L) regions have some affinity for the determinants of the antigen molecule. Subsequent release of Ag from the (Ag.H.L)(n) complexes yields free antigen and antibody molecules. Each of the released Ag molecules can be used repeatedly for the same reaction cycle and thus can induce the biosynthesis of a large number of antibody molecules. Any macromolecule, natural or synthetic, that has at least a few polar groups and that can penetrate to the nascent H and L chains can thus act as an antigen. Whereas the structure of the H and L chains is genetically determined and transmitted through the germ line, the process induced by the antigen is a phenotypic phenomenon. The antigen acts in this process as a stereospecific cofactor or regulator of the thiol-disulfide transhydrogenation of the combining H and L chains of immunoglobulins.


Proximal versus peripheral hepatic artery embolization experimental study in monkeys. A case is described in which extensive embolization of the hepatic artery to reduce insulin production by a metastatic islet-cell tumor resulted in liver failure, intrahepatic abscesses, and death. The difference between proximal and peripheral hepatic artery obstruction was investigated in monkeys by embolizing hepatic arterial beds with Gelfoam and silicone. Gelfoam obstructed proximally, arterial collaterals developed rapidly, and liver function remained normal. Peripheral hepatic artery embolization with silicone produced liver infarction and severe functional abnormalities. Peripheral hepatic artery occlusion was more effective in preventing the development of collateral circulation but involved a significant risk of hepatic failure or abscess.


Neuropathic injuries to the lower extremities in children. Neuropathic injuries of the lower extremities in children due to meningomyelocele, congenital indifference to pain, and peripheral nerve damage were studied in 10 patients. The injuries fell into four categories: (a) fractures of the metaphysis and diaphysis of long bones, (b) epiphyseal separation, (c) Charcot joints, and (d) soft-tissue ulceration. These injuries are often unrecognized; untreated, they can lead to severe disability. For patients with impaired sensation, radiographs should be obtained at any sign of localized soft-tissue swelling, warmth, or hyperemia, especially near a joint. Following diagnosis, immobilization of the limb will lead to prompt healing of fractures and epiphyseal separation.


Studies on the antigenic structure of Bacteroides nodosus. The antigenic mosaics of three Bacteroides nodosus isolates (198, 199 and 127) were studied to elucidate the nature of the protective immunogen. In vaccinated sheep the three isolates induced high homologous serum agglutinin titres but it was also apparent that 198 and 199 shared a major surface antigen not present on 217. This major cross-reacting antigen was not detected with rabbit antisera. The fimbriae, consisting predominantly of protein, induced high homologous titres in rabbits and represented the type-specific antigen. Lipopolysaccharide (LPS) from each of the isolates induced low agglutinin titres and high 2-mercaptoethanol-sensitive indirect haemagglutinating antibody titres. The heat-stable LPS contained at least two common carbohydrate O antigen determinants but no type-specific O antigens were detected.


Anaphylaxis in isolated rabbit lungs.

Anaphylaxis in isolated rabbit lungs. Lungs from rabbits sensitized to ovalbumin or bovine gamma-globulin were isolated and perfused with autologous blood. The response to antigen challenge via the perfusate was immunologically specific and characterized by a marked increase in perfusion resistance, a moderate increase in airway resistance and a small decrease in lung compliance. The response could also be elicited by specific antigen challenge in lungs from sensitized rabbits perfused with blood from normal rabbits and in lungs from normal rabbits perfused with blood or plasma from sensitized rabbits. The magnitude of the response was greater when blood or plasma from sensitized animals was used as the perfusate. Therefore, blood and/or plasma factors appear to be the major contributors to the response.


[Ontogeny of the immune response in mice. I. Presence of IgG and alpha fetoprotein in the body fluids of mice of various ages]. Alpha protein and IgG were determined by radial immunodiffusion in Swiss mice, in the amniotic fluid, milk and serum of the mice at different ages (fetus, neonates, at the age of 5, 10, 15, 20, 25, 30, 35 and 40 days and adults). The highest alpha-protein concentrations were found in the fetal serum (0.20 mg/ml) ; after birth serum levels rapidly fell and at 20 days could no longer be detected ; it was present in the amniotic fluid and milk collected within the first 24 hours after birth. In contrast, IgG was found in small concentrations in the fetal serum (0.60 mg/ml) but increased with age after birth. This immunoglobulin was present in the amniotic fluid and mothers' milk for 15 days. The significance of the results is discussed.


The repertoire of antibodies to a hapten group in an inbred strain of mouse. We have investigated the diversity of antibodies to the p-azophenylarsonate (Ar) hapten group in the A strain of mouse. The method used consisted of suppressing an idiotype which commonly is observed in the anti-Ar antibodies of all this strain, and then generating anti-idiotypic antisera against the anti-Ar antibodies (lacking the cross-reactive idiotype) which arose after hyperimmunization of the suppressed mice. Anti-idiotypic antibodies prepared against these anti-Ar antibodies were used as a tool to investigate the frequency of recurrence of individual idiotypes in other mice of the same strain. A highly sensitive radioimmunoassay indicated that 3 of the 4 idiotypes studied were virtually undetectable in a panel of 181 A/J mice immunized against the phenylarsonate hapten. A fourth idiotype was found but at very low frequency. The results emphasize the enormous diversity of the immune response to a single hapten group within a strain of mouse. They are interpreted on the basis of hypothesis that the diversity arises through a random series of somatic events. The common cross-reactive idiotype, which is present in the anti-Ar antibodies of all normal A/J mice, is postulated to be the product of a germ-line gene which is closely related to a germ-line gene through a small number of somatic mutations which takes place in every A/J mouse.


Use of mitogens for the functional characteristization of human lymphocyte subpopulations with special reference to activation of human B lymphocytes by antibodies to beta2microglobulin. The paper describes the use of mitogens for the function characterization of human lymphocyte subpopulations with special reference to the activation of human B lymphocytes by antibodies to beta2microglobulin.


Typing for MLC determinants: methods and applications. MLC typing of random individuals can be performed using a panel of inactivated HLA-D homozygous cells. Eight different HLA-D specificities are now internationally accepted. The evaluation of the results must take into account both the general responding capacity of the cell to be typed and the general stimulating capacity of the typing cells. An evaluation based on the 75th percentile is discussed in detail and some pitfalls are mentioned. Furthermore a description is given of primed lymphocyte typing (PLT), where cells primed in ordinary MLC cultures to one HLA-D determinant have the ability to respond in an accelerated way to similar HLA-D antigens when re-exposed to such cells in secondary cultures. In our experiments, an excellent correlation is found between these two ways of MLC typing provided that the cells used for priming are well characterized (i.e. HLA-D homozygous cells). Finally, some clinical applications of MLC typing are described, especially in connection with transplantation and association between HLA and various diseases.


Increased serum alpha-fetoprotein levels in cytomegalovirus infections. Among 54 patients with a clinically and serologically verified cytomegalovirus infection, 8 (15%) had raised alpha-fetoprotein (AFP) levels in sera taken after the onset of infection. No raised serum AFP was seen in acute or convalescent sera of 45 patients with other viral diseases, including infectious mononucleosis and acute herpes simplex virus infection. No definite correlation was found between raised AFP and raised S-ALAT values, indicating hepatic damage. The highest single AFP values were seen in two immunosuppressed patients and in a patient who received a blood transfusion. Two possible explanations for this phenomenon are advanced.


Collagenase in synovial fluid.

Collagenase in synovial fluid. Free collagenase activity was demonstrated in all 13 normal, 5 osteoarthrotic and 54 rheumatoid synovial fluids studied. Osteoarthrotic, normal, and rheumatoid fluids showed a free collagenase content increasing in that order. However, the total activities after trypsin activation did not differ significantly. The concentrations of alpha1-at and alpha2-M in synovial fluids reflected those in sera. The complex interactions between serum inhibitors, and free or latent collagenase released from both synovial cells and leukocytes are discussed.


Thyroid hormones homeostasis and TSH in autonomic thyroid nodule. Thirty patients with autonomic thyroid nodules were studied. Thyroid scintigram, iodine uptake at 2 and 24 hrs, PBI, total thyroxine, serum T3(RIA), basal TSH and 30 minutes after TRH administration were performed. A lack of concordance was found between the clinical signs and the laboratory data. According to the data obtained the patients were grouped into 3 categories: having 1) autonomic thyroid nodules with normal PBI, T4t, T3 values and absent or diminished TSH response to TRH; 2) autonomic thyroid nodules with increased T3 secretion and no response to TRH despite a normal T4 and iodine uptake, 3) hyperfunctional autonomic nodules in which PBI, T4t, T3 and iodine uptake are increased. TSH response to TRH is the most sensitive test for hyperthyroidism, more sensitive than the circulating thyroid hormones assay, and best fitted for adequate therapeutic management. Serum triiodothyronine was found to be increased in 95% of the patients. Thus, there is a T3 thyreotoxicosis in most of the autonomic nodules. TRH test and serum T3 assay being essential elements in starting the treatment, they have to be included among the tests used in autonomic thyroid nodules investigation.


Iodide-trapping defect of the thyroid. A case report. We describe a grossly hypothyroid 50-year-old woman, mentally retarded since birth. On the basis of her history of recurrent goitre, absence of 131I neck uptake and a low saliva/plasma 131I ratio, congenital hypothyroidism due to a defect of the iodide-trapping mechanism was diagnosed. Other family members studied did not have the defect.


alpha-Fetoprotein in liver disease.

alpha-Fetoprotein in liver disease. Ther serum concentration of alpha-fetoprotein (AFP) was measured by radio-immunoassay in 98 patients with liver disease including hepatoma, chronic active hepatitis, alcoholic cirrhosis, and acute virus B hepatitis. Raised AFP levels, above 30 ng/ml, were found in 87% of patients with acute viral hepatitis, in 82% of patients with primary liver cell carcinoma, in 58% with chronic active hepatitis and in 14% of patients with alcoholic cirrhosis. However, levels above 1 000 ng/ml were found only in patients with hepatoma and in acute viral hepatitis.


Liver tumours associated with the use of contraceptive pills. The possible association between benign liver tumours and the use of oral contraceptives is reviewed. The possible aetiological role of the pill, the confusion regarding the pathology of these lesions, and the presentation and diagnosis of the 4 clinical categories are discussed. These categories are: an incidental, asymptomatic lesion; a mass in the abdomen; upper abdominal pain; and shock due to rupture, with intraperitoneal haemorrhage. The importance of an awareness of the condition and the use of liver scanning, ultrasound and hepatic angiography in making a diagnosis, are emphasized. Biopsy is essential to establish the diagnosis and to differentiate benign from malignant tumours. A policy of treatment for the patients with different clinical presentations is detailed.


beta-thromboglobulin and deep vein thrombosis. The measurement of plasma beta-thromboglobulin as a potential diagnostic test for venous thrombosis has been investigated in 16 normal volunteers, 24 patients presenting with deep vein thrombosis (DVT) or pulmonary embolism and 46 patients screened by 125I fibrinogen test (IFT) for post-operative DVT. The normal mean was 33 ng/ml (range 15-117 ng/ml). Of the 24 patients with clinical thrombotic disease 22 presented with DVT confirmed by phlebogram or IFT and 2 presented with embolism confirmed by lung scan. At the time of first presentation 12 out of 24 had betaTG values greater than 70 ng/ml. All except 3 of this group of 24 patients had values of greater than 70 ng/ml at some stage during a subsequent week of daily sampling. DVT was detected in 13 out of 46 screened post-operative patients. There was a rise om betaTG observed within 24 hr of the IFT becoming positive but the mean rise did not reach significance at the 5% level. An association between DVT and high betaTG values has been confirmed. However, its clinical value cannot yet be fully elucidated until factors, probably related to blood sampling and clearance, are further investigated.


The measurement of human plasma antiplasmin activity by radial diffusion assay. An assay of human antiplasmins has been developed utilizing radial diffusion of plasma from wells cut in plasmin-enriched, fibrinogen-agarose plates. After diffusion the fibrinogen is clotted. Zones of fibrin protected from background fibrinolysis develop as the result of plasma antiplasmin activity. A pooled plasma standard was taken to contain 100% antiplasmin activity. Antiplasmin activity of 52 normal subjects varied from 64 to 132%. Washed platelets contained 1-5% antiplasmin activity. Using antisera to precipitate individual inhibitors, physical methods of separation, and electrophoresis of plasma in agarose, several different proteins were found to have antiplasmin activity in this assay. Thus, alpha2-macroglobulin contributed 56%, alpha1-antitrypsin 20%, antithrombin III 2%, and other proteins 22% of the total antiplasmin activity. 1 ml of whole plasma neutralized 7.0 CTA units of plasmin.


Nucleoprotein localization by bismuth staining. Experiments on isolated mouse liver muclei involving enzyme digestion, the crosslinking of amino groups and alkaline hydrolysis demonstrate that bismuth binds to nucleoproteins through amino and phosphate groups. Analysis of the nucleoproteins extracted with salt and acid solutions in conjunction with bismuth staining after these treatments suggests that: (1) a bismuth amino group interaction occurs on ribonucleo-protein particles, histones and perhaps some non-histone chromosomal proteins, and (2) bismuth phosphate binding is specific for one, or all, of three distinct species of non-histone proteins. These results suggest that histones not tightly bound to DNA through their amino groups are present on interchromatin granules, the presumed transcriptionally active regions of chromatin. Phosphorylated non-histone proteins are also localized at these sites. Staining with heavy metals such as bismuth may be the best method for high resolution localization of nucleoproteins involved with regulating gene activity and maintaining chromatin structure.


Indirect cholangiojejunostomy using a Latex T tube for palliation of malignant jaundice. Indirect cholangiojejunostomy was carried out using a Latex T tube for palliation of non-curative malignant jaundice and its technique is described. The surgical procedures reported here can separate cholangiojejunostomy which otherwise involves some difficulty into three simple steps: 1) insertion of a Latex T tube into the common bile duct or hepatic duct, 2) Witzel type internal fistulization, and 3) Roux-en-Y jejunostomy. The reported technique can be one of the useful arms for control of obstructive jaundice due to malignancy of the biliary-pancreatic region and may have a special appeal to general practitioners.


Effects of lindane upon reproductive function in a 3-generation study in rats. A 3-generation study, involving the feeding of lindane at dietary concentrations of 25, 50 or 100 ppm to CD strain rats, did not reveal any adverse effects upon reproductive function as compared with that of control animals. There were no major malformations, while the distribution of minor variants was not compound or dose-related. An examination at 21 days of age of 10 males and 10 females F3B animals in each group revealed a dosage related tendency for increased liver weight and enlarged hepatocytes were seen in some control and treated animals. The relevance of these latter findings was considered of doubtful importance compared with the lack of effects on the growth and reproductive performance of the preceding generations.


Histopathological and biochemical changes in guinea pigs after repeated dermal exposure to benzene hexachloride. Dermal application of benzene hexachloride in daily doses of 100, 200 and 500 mg/kg for a total period of 30 days caused significant changes in male guinea pigs. The animals exposed to high doses of benzene hexachloride (1,2,3,4,5,6-hexachlorocyclohexane) (BHC) (BHC) died within 5--12 days. There was no mortality in 100 mg/kg/day but significant pathologic and biochemical changes were observed in the vital organs of the experimental animals. Massive congestion and thickened blood vessels were seen in liver of the BHC treated animals in comparison to the normal picture of the controls. Similarly, testicular changes included mild to severe pathologic lesions. There was no change in the epididymis, kidney, spleen, brain and lungs. The changes in the skin were mild and no signs of dermatitis were observed in the BHC painted areas. The activity of glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transaminase (GPT) and alkaline phosphatase in liver and serum revealed significant changes from that of the controls. The significance of biochemical changes with the tissue damage of the insecticide exposed animals are discussed.


Is the presence of serologically defined target cell antigens sufficient for binding of cytotoxic T lymphocytes? Target cell parameters that coincide with specific binding (conjugation) of target cells to cytotoxic T lymphocytes were examined. It was demonstrated that concentrations of formaldehyde (0 to 1.2%) which do not affect the capacity of target cells to exclude dyes and incorporate sodium chromate, or their susceptibility to cellular and humoral immune lysis did not affect target cell conjugation. Treatments with higher formaldehyde concentrations (1.6 to 3.2%) caused similar decreases in all of the above-mentioned target cell parameters. Formaldehyde-treated (over 0.1%) targets that could not cap alloantigens or incorporate macromolecular precursors could still conjugate with cytotoxic T lymphocytes. Quantitative changes of serologically detected antigens alone cannot account for the decrease in binding of formaldehyde (greater than 1.2%)-treated target cells to cytotoxic T lymphocytes, since absorption of lytic alloantibodies by formaldehyde-fixed targets was not affected over the fixative range used (0 to 3.2%). From this study, it appears that certain target cell properties related to membrane structure and function and not the apparent expression of serologically defined antigens alone are essential for binding of targets by cytotoxic T lymphocytes. Additionally, it was shown that target cell susceptibility to immune lysis is retained, as long as their ability to bind to cytotoxic T lymphocytes is not affected.


Xenoantibody directed against molecular components of the HLA system. Quantitation of antibody to serologically defined HLA complexes on the cell surface. A quantitative absorption-blocking assay was used to measure the levels of antibody in immune rabbit sera that react with the serologically defined HLA molecular complex on the cell surface. Quantitation was determined by comparing the number of xenoantibody-treated cells with the number of untreated control cells needed to absorb a defined amount of cytolytic alloantibody. Complement-dependent cytolytic titers were used to standardize the total antibody content reactive with cell surface antigens of xenoantisera used to treat absorbing cells. The ability of xenoantibody to "block" or inhibit the reaction of alloantibody with the absorbing cell was observed to be the result of concomitant removal of both xeno- and alloantigenic determinants from the cell surface. Immunofluorescence data with xenoantibody suggested that removal was accomplished by temperature-dependent selective redistribution and endocytosis of the serologically defined HLA molecule complex. By using these procedures, xenoantisera have been characterized that contain different amounts of antibody to the two-component HLA complex relative to their total cytolytic antibody content. Variation ranges from a xenoantiserum with little or no blocking capacity to several highly effective serologically defined HLA blocking xenoantisera.


Immunobiology of the graft-versus-host reaction. I. Symptoms of graft-versus-host disease in mice are preceded by delayed-type hypersensitivity to host histocompatibility antigens. During initiation of an acute graft-versus-host reaction by injection of C57BL/Rij spleen cells into lethally irradiated (C57BL/Rij X CBA/Rij)F1 hybrid mice, a state of delayed-type hypersensitivity (DTH) against host histocompatibility (H) antigens occurs. This was demonstrated by means of transfer of host spleen and lymph node cells into C57BL/Rij recipients, which received a challenge with CBA/Rij spleen cells. Initiation and transfer of the graft-versus-host-related DTH reactivity was highly dependent on Thy-1.2+ cells. The development of DTH reactivity started between 8 and 24 hr after semiallogeneic spleen cell transplantation and increased during the days thereafter. In the spleen maximal DTH reactivity was found on day 4, whereas in the lymph nodes maximal reactivity occurred on day 5 after irradiation and reconstitution. Thereafter, the reactivity decreased until there was no further DTH reactivity demonstrable on day 13. The specificity of the DTH reactivity for host H antigens was proved by no reactivity to a challenge of DBA/2 and Swiss spleen cells, which are H-2-incompatible with CBA cells.


Serum alpha 2 globulin concentrations in South African Negro adults free from coronary heart disease. In Western populations serum alpha 2 globulin concentration has been reported to be significantly higher in patients with coronary heart disease (CHD) compared with those unaffected. In South Africa, since CHD is extremely rare in Negroes in rural areas, it was judged of value to determine serum alpha 2 globulin concentrations in I. groups of rural elderly Negroes (men and women), and, for comparison, II. two groups of middle-aged to elderly White men, one free from overt CHD, and the other, patients with CHD. Mean values on the Negroes were significantly higher than those on both the White groups. In the latter, the group with CHD had a significantly higher mean value than the non-CHD group. Possible reasons for high values in the Negro groups are an ethnic difference, early malnutrition, greater exposure to infections, and pattern of habitual diet. It is concluded that interpretation of a raised serum alpha 2 globulin concentration in relation to proneness to CHD is influenced by the population concerned.


Simultaneous determination of human sperm morphology and viability: simple office technique. Determinations of sperm morphology and viability are useful techniques for assessment of semen quality. In this report eosin-nigrosin is compared with hematoxylin-eosin for effectiveness in characterizing spermatozoa. The results indicate that eosin-nigrosin and hematoxylineosin give comparable results in differentiating spermatozoa. In addition, eosin-nigrosin allows the determination of the per cent of viable cells in each morphologic category. The eosin-nigrosin staining technique is a simple, rapid method which may give additional information in the assessment of male infertility.


Red cell ABH antigens in leukaemias and lymphomas. The qualitative and quantitative determinations of A, B, and H antigens were studied in blood samples of 509 normal subjects and 114 patients. 11 of 53 (20.75%), 3 of 27 (11.11%), and 4 of 34 (11.76%) of adults with leukaemia, children with leukaemia and adults with lymphomas, respectively, show significantly lower antigenic scores than normal controls. Since all A, B and H antigens were found to be independently affected, the destruction of well-developed ABH antigens by a certain substance produced during the course of the disease was suspected.


[Cardiac arrhythmia during pneumatic dilatation of achalasia of the esophagus (author's transl)]. ECG was recorded in 38 patients with achalasia of the esophagus before, during and after pneumatic dilatation. During the dilatation both marked increases as well as decreases of heart frequency were observed. The mean values of all patients remained virtually unchanged. The most frequent type of arrhythmia were ventricular extrasystolies (16 patients), whereas supraventricular extrasystolies (4 patients), second degree-av-block (2 patients) and av-dissociation (1 patient) occured infrequently. In none of our patients arrhythmia was critical, even in a patient with preexisting cardiac insufficiency. Thus, patients with achalasia of the esophagus are not endangered to a major degree by cardiac arrhythmia during pneumatic dilatation.


Investigations on the efficacy of two schemes of standard gammaglobulin administration in the prevention of viral hepatitis. Prophylactic administration of standard gammaglobulin (G) to over 5,000 children led to a more than 6-fold decrease in the incidence of icteric viral hepatitis (V.H.) as compared with the control group and to the disappearance of the autumn--winter epidemic peak. A 3-fold decrease in the incidence of subclinical forms was also achieved. In another group of 5,000 children G was only given to contacts in epidemic foci; the incidence of icteric V. H. was similar to that recorded in controls, while subclinical forms were 2 times more frequent in children having received G after the infectant contact.


[Lantern slides (author's transl)].

[Lantern slides (author's transl)]. Bad projection of lantern slides impairs every paper. Causes are incorrect conception of the slide, insufficient projection. The conditions of optimal projection are discussed in detail (projector, screen, room-image factor, optics, lay out of slides).


[Effect of oral contraceptives on the psyche and on sexuality]. The problem of whether or not oral contraceptives affect the psychic function of the female is still controversal. The purpose of the present investigation was to study the occurrence of psychic and sexual effects in women taking either Ovosiston (mestranol 0,08 mg + chlormadinone acetate 2, 0 mg) or Non-Ovlon (ethinylestradiol 0,05 mg + norethisterone acetate 1,0 mg). 186 somatically healthy women were subject to two interviews, immediately before the start of therapy and six months after it. Most of the patients recorded distinctly alterations of their mood, their initiative, their elemental desires such as appetite, sleep, need of warmth, and their sexual behaviour. 73 of them noted a change in their emotional pattern, which was positive in 34 and negative in 39 cases. 99 women reported changes in their elemental desires. 136 patients exhibited alterations of their sexual behaviour, which were assessed as an improvement or impairment in 108 and 28 cases, respectively. There is a statistically significant correlation between the impairment of sexual function and the incidence of other untoward side effects. On the other hand, negative suggestion by poor experience of other women on by the influence of mass media seems to play a minor role in producing changes of sexual behaviour. There is, however, a clearcut relationship between negative suggestion and the incidence of unfavourable emotional alterations. The findings are discussed with reference to their clinical significance.


[Detection of antigen of rickettsia of the tick group in bird droppings by antibody neutralization test]. The author presents methodical materials pointing to a possibility of using a micromethod of the antibody neutralization test for detection in bird pellets of the antigen of rickettsia referred to the Rocky Mountains spotted fever tick group. Specificity of the test, rickettsia antigen and sera was studied. The method is recommended for detection at the territory under study of circulation of the causative agent of tick-borne rickettsiosis by revealing the specific antigen in bird pellets.


[Immunoelectrophoretic analysis of the antigenic structure of Sh. sonnei]. The authors studied the antigenic composition of 105 Sh. sonnei strains freshly isolated from patients suffering from acute dysentery and carriers. Immunophoregrams of pure S-and R-forms species were obtained. Up to 13 antigens differing by electrophoretic and diffusion mobility and immunological specificity were revealed among soluble Sh. sonnei antigens The position of common and specific antigens was determined on the immunophoregram. Along with the thermostable somatic O-antigen detected at the I phase of the S-forms, and two thermolabile O-antigen components at the II phase, and the R-forms, there was revealed a surface, relatively thermolabile, K-antigen of A-type capable of agglutinating live bacteria in the O-antiserum; position of the latter on the immunophoregram was also determined.


[Experience in studying protein A in staphylococcal cultures]. Methods of quantitative and qualitative determination of protein A in staphylococcus cultures were studied comparatively. The maximal number of strains positive by protein A were revealed by means of indirect hemagglutination test with cell extracts. Quantitative and qualitative characteristics by protein A can be used in studying the problems of strain and clone heterogeneity of S. aureus. A common antigen was revealed in the reactions with human gamma-globulin in 21 of 38 S. aureus strains, and in 1 of 11 S. epidermidis strains; the presence of this antigen failed to correlate with the quantitative protein A content in the same strains.


Histochemical observations on the giant neurosecretory cells of the thoracic ganglion of the adult and juvenile crabs, Potamon magnum magnum (Pretzman). The giant neurosecretory cells in the thoracic ganglion of the adult and juvenile crab, Potamon magnum magnum (Pretzman) were histochemically investigated. The secretion is mainly proteinaceous in nature, containing considerable amounts of acid mucosubstances, sulphate esters, lipids and a little carbohydrate but no glycogen. The detailed nature of proteinaceous neurosecretory material in the adult crab was further tested. It appears that the neurosecretory material of these cells contains moderate amounts of sulfhydryl groups and few of disulphide bonds. No trace of tyrosine could be observed. The neurosecretory granules were associated with considerable amounts of cytoplasmic RNA. In general, stronger reactions were obtained in summer and winter than in other seasons.


Effects of verapamil in ventricular premature beats of acute myocardial infarction. In 8 patients affected with acute myocardial infarction verapamil was highly effective in controlling ventricular extrasystoles. Theoretically the drug ought not to be effective if the extrasystoles were related to fast cardiac fibers characterized by a rapid inward Na+ current. The result suggests that in acute myocardial ischemia ventricular extrasystoles are related to slow inward Ca++ currents.


Pancreatic duct glands. I. Staining reactions of acid glycoprotein secret. The staining properties of secretory granules present in pancreatic duct glands and in secretory cells of pancreatic duct epithelia are studied in cat, dog, and man. The secretory material contains histochemically detectable disulfide groups and tryptophan. The mucosubstance is characterized as GC-mucin B (2.5) A (0.8 MgCl2), in part N (i.e. a sialoglycoprotein with vic-glycols, sialic acid carboxyls, being basophil at pH = 2.5, alcianophil in the presence of 0.8 M MgCl2, and in part sensitive to digestion with alpha-neuraminidase).


Toluidine blue staining of cartilage proteoglycan subunits. Selective binding of toluidine blue to basophilic proteoglycans is the first stage of a staining method which proceeds to the formation of a heavy metal salt of the dyestuff. By means of this procedure matrix complexes of hyalin cartilage were stained such as to display granular subunits about 25 to 35 A in diameter. The findings are interpreted as the demonstration of proteoglycan constituents of chondromucin aggregates.


Multiple sclerosis specific antigens in MS brains. Antigens in MS and non-MS brains were investigated by means of crossed immunoelectrophoresis. Two MS specific antigens were found: a measles antigen which was present in microsomes and cytosole, and an unidentified antigen present only in cytosole.


Luxol-fast-blue-staining of nerve cell nucleoli as indicator for acidotic lesion in cases of intracranial hypertension? The allochromatic behaviour of nerve cell nucleoli in the Luxol-fast-blue-method (blue nucleoli) in cases of raised brain pressure is probably due to a lactacidotic pH-shift basing on cerebral hypoxia. In 109 out of 134 cases with blue nucleoli signs of cerebral hypertension could be observed. The IEP of nerve cell nucleoli was determined in the nuclei pontis. Clinical and pathophysiological data were evaluated.


Inhibitory effects of imidazolines on histamine liberation from human leukocytes and on tracheal smooth muscle tone. Antigen-induced IgE-mediated release of histamine from human leukocytes, an in vitro model of allergic reactions, was blocked by imidazole and imidazole-compounds such as oxymetazoline and clonidine. The H-2-antihistamines antagonized this effect of imidazolines. Alpha- and H-1-receptor blocking agents did not antagonize the effect. The contractile effects of the imidazolines were tested on tracheal preparations from the cow and guinea-pig. Imidazole was found to be a rather potent contracting agent, while oxymetazoline only caused weak contractions. Clonidine relaxed the tracheal muscles, when used in the concentration range which were inhibitory in the leukocyte experiments. The contractions caused by imidazolines were non-competitively inhibited by clemastine, while the relaxing effects were blocked by a combination of propranolol, phentolamine and cimethidine. The results suggest that imidazolines which inhibit histamine release and relax bronchial smooth muscles may be of therapeutic importance in the treatment of human allergic disorders.


Absence of C-type virus production in human leukemic B cell, T cell and null cell lines. Electron microscope observation of cultured human leukemic B cell, T cell and null cell lines and reverse transcriptase assay of the culture supernatants were all negative for the presence of C-type virus. Bat cell line, which propagates primate C-type viruses well, was cocultivated with the human leukemic cell lines, in the hope of amplification of virus if present. Three weeks after mixed culture, the culture supernatants were again examined for reverse transcriptase activity and the cells were tested for syncytia formation by cocultivation with rat XC, human KC and RSb cell lines. All these tests, except for the positive control using a simian sarcoma virus, were negative, suggesting that no C-type was produced from these human leukemic cell lines.


Descending course of primary afferent collaterals in the cat's spinal cord. After injecting horseradish-peroxidase into the lower thoracic, lumbar and sacral spinal cord segments of cats, labelled perikarya were found in several spinal ganglia localized cranially to the site of injection. The segmental distance between the injection site and the rostralmost localized ganglion with labelled cells varied depending on the medio-lateral localization of the injection. The longest distance (10 segments) was achieved by medial injections. Primary sensory neurones with long descending collaterals belong to large ganglion cells.


[Anterior and posterior medullary analgesic stimulation, using a percutaneous implantation technic]. The technique of percutaneous implantation of a pain-relieving stimulator reduces the surgical procedure considerably. This advantage is, however, lessened by the absence of precision in placing epidural electrodes, and by the risk of their later displacement leading to inadequate stimulation. To reduce this disadvantage the authors suggest a technique that is a modification of the classical open procedure: it is particularly suitable for implantation at the cervico-dorsal junction. In 12 patients treated by percutaneous implantation, 5 received, from epidural electrodes, an anterior medullary electrical stimulation which caused analgesia without paraesthesiae in the painful ares. The results, however, do not justify a conclusion that anterior medullary stimulation is better than posterior column stimulation.


Thyroid function in primary depression.

Thyroid function in primary depression. The thyroid function of 20 female inpatients with primary depression was investigated by the following tests: serum thyroxine, in vitro radioactive triiodothyronine uptake, free-thyroxine index, thyroidal 131I uptake and 48 hours protein-bound 131I. The mean values of serum thyroxine, free-thyroxine index and thyroidal 131I uptake of the patients were found decreased, compared to those of the euthyroid controls (to a statistically significant level for serum thyroxine and free-thyroxine index P less than .001). These findings point to a decreased thyroid activity in primary depression. The possibility of implication of a common neurochemical mechanism in both primary depression and thyroid hypoactivity was discussed.


[Development of a standard model of nutritional liver injuries which could be used in experimental pharmacology]. This work was undertaken to elaborate a standard model of nutritional liver injury similar to that occurring in the deficiency states in human pathology with regard to functional troubles and morphological lesions. Rats of the Wistar strain were fed two low protein diets (diet A and diet B) containing however the full composition of vitamins and minerals. The diet A showed the most advantages as it provoked mild liver changes; but it proved to induce continuous impairment of the liver function in parallel direction with the parenchymal lesions. The full correlation of the morphological lesions and the functional disturbances observed in experimental animals as well as their intimate ressemblance to the clinical picture and hepatic lesions encountered in the deficiency states in human pathology suggest that present model of nutritional liver injury could be useful in experimental pharmacology.


A colloidal iron silver impregnation method for acid mucopolysaccharide. A simple and rapid method for the demonstration of acid mucopolysaccharide using colloidal iron, sulphosalicylic acid ammoniacal silver is described. Satisfactory staining of human as well as animal tissue was obtained following 10% neutral formol saline fixation and paraffin sections. There were no oxidation or hydrolysis sequences. The staining specificity was controlled using methylation and saponification. The distribution of the acid mucopolysaccharide demonstrated by this method was identical with that obtained by colloidal iron Perls' reaction.


[Detection of cell-mediated immunity against protein antigen by the macrophage adherence inhibition test (author's transl)]. The macrophage adherence inhibition test is a suitable assay for detection of cell-mediated immunity against bovine gamma globulin (BGG) in guinea pigs. The test was performed with peritoneal cells (PC) in hemocytometer chambers. 10 mg/ml BGG were added as antigen. The washing step was performed with a burette. An average standard deviation of 5% was obtained with this washing method. Day-to -day-investigations in individual animals show differences with respect to the degree of cell-mediated immunity against BGG. Supernatants of sensitized spleen cells or lymph node cells produce adherence inhibition in PC of nonimmunized guinea pigs too.


Localization of human pancreatic polypeptide in an argyrophilic fourth cell type in islets of the rat pancreas. An argyrophilic fourth cell type in the rat pancreatic islet can be differentiated from other silver-staining cells by using a modification of the Grimelius aqueous silver nitrate technique. Restaining of the tissues using fluorescent techniques with anti-HPP (Human Pancreatic Polypeptide) serum results in bright fluorescence in the fourth cell type.


Structural differences of pale and strongly stained motor end-plates of the rat diaphragm. According to the staining intensities for AChE, the motor end-plates of the rat diaphragm can be classified into strong (S) and pale (P) types. About 34% of the total end-plates of the rat diaphragm are of S type and 50% of P type. The P end-plates differ from S end-plates in two aspects. First, the secondary subneural clefts of the S end-plates are well developed. They are numerous, long, closely packed and often branched. On the other hand, the secondary subneural clefts of the P end-plates are short, sparse and usually unbranched. Secondly, there seems to be a variation in AChE activity in the P end-plates. Focal negative AChE areas are found in the subneural apparatus of some P end-plates. It is concluded that the less well developed secondary subneural clefts and focal areas of negative AChE activity contribute to the paler staining of the P end-plates.


Fetal-maternal distribution of ovine alpha-fetoprotein. The concentration of ovine alpha-fetoprotein (AFP) was determined in sheep fetal, maternal, and neonatal sera and amniotic fluid by radioimmunoassay. The fetal serum AFP concentration was highest during the first third of pregnancy and continued to decline with fetal and neonatal development. Total fetal synthesis of AFP was highest during the late middle and early latter third of pregnancy. Amniotic fluid AFP concentration was highest during the early middle third of pregnancy. Maternal serum AFP was not elevated above nonpregnant control levels during the first two-thirds of pregnancy, but instead showed a tendency to be elevated only during the last third of pregnancy. The fetal physiological distributions of ovine and human AFP in fetal serum and amniotic fluid appear to be similar, whereas in maternal serum the pattern of AFP levels differs in the two species as a function of gestational age.


Histochemical demonstration of disulfide-groups in the lamina propria of human seminiferous tubules. The distribution of disulfide-groups was investigated in the tunica propria of human seminifersou tubules by means of a thiosulfation/Alcian Blue + 0.8 Mol MgCl2-staining reaction. Controls had shown the absence of significant amounts of sulfhydryl- or sulfate-groups in the lamina propria, which groups would also be demonstrated by the method employed. The lamina propria of human seminiferous tubules is rich in disulfide groups. The staining reaction decreases in the region of the tubulus rectus, is only faint in the connective tissue which underlies the epithelium of the rete testis, and is absent in the lamina propria of efferent ducts. It is suggested that microfibrils and type IV collagen (both rich in cystine) are the materials responsible for the histochemical reaction described. The occurrence of multiple layers of basal lamina material (type IV collagen) and bundles of microfibrils is shown in comparative electron microscopic studies.


Golgi study on cat hippocampal formation. The Golgi architecture of the Fascia dentata and hippocampus is described in the cat. The main cell types are like those found in other species. The initial collaterals of granule cell axons were demonstrated and are commented on. The interneurons proved to be of several types and subtypes not observed in lower species. The intrinsic fibers and few of the afferents -- both in the dentate fascia and in the hippocampus -- are discussed.


Some observations of mycobacterial acid-fastness. Mycobacterial acid-fastness seems most readily explainable on the basis of a "lipids barrier principle" that in the first instance hinders penetration of the water-soluble dyestuff, and in the second, hinders penetration of the bleaching acid. Mycolic acids, and more especially, mycolate salts, in xylene solution complex and solubilize crystal violet, but the complex is demonstrably not stable even to weak acids. Stained mycobacterial preparations are readily bleached by an acidified lipid solvent; the destained preparations are nevertheless contribution to the barrier on which acid-fastness depends appears to reside in the mycoloyl-arabinogalactan of the cell wall.


Controlled clinical trial of four short-course regimens of chemotherapy for two durations in the treatment of pulmonary tuberculosis: first report: Third East African/British Medical Research Councils study. A comparison was made between 4 regimens of chemotherapy given for 6 or 8 months, with a 6-month period of follow-up thus far. One regimen, which had a 2-month initial intensive phase of streptomycin, isoniazid, rifampin, and pyrazinamide followed by daily administration of thiacetazone and isoniazid, was very effective. Shortening the initial intensive 4-drug phase to 1 month produced a less successful regimen unless the continuation phase was twice-weekly administration of streptomycin, isoniazid, and pyrazinamide. A regimen in which pyrazinamide was omitted from the initial intensive 2-month phase was also less effective. All patients were inpatients for 6 months to ensure that every dose of drug was fully supervised during this period. Increasing the total duration of chemotherapy from 6 months to 8 months was associated with a significant decrease in the early relapse rate in all 4 regimens, although the above differences between the regimen remained. The applicability of the findings is discussed.


[Cellular energy metabolism in rats receiving diets contaminated with lindane]. Young male rats, Wistar CF strain, about 70 g body weight, were fed a well-balanced diet containing 0 (control), 60 or 240 ppm lindane. The day before the experiment, all the animals were fasted, and some of them placed in a restraint wheel forcing them to walk on during 18 hrs; another group was given an i.p. injection of 2.6 g/kg glucose 30 minutes before their sacrifice. The redox and energy potentials of liver and muscle tissues were estimated after the determination of the following compounds: lactate, pyruvate, beta-hydroxybutyrate, acetoacetate, ATP, ADP, AMP, inorganic phosphate, NADP and NADPH. No effect of lindane was observed on muscle metabolism and the 60 ppm dose was without significant effect on liver metabolism. At the 240 ppm dosage: a. Lindane ingestion increased the liver betaHOB/AcAc ratio and decreased the Lac/pyr ratio. The ATP/ADP ratio was not significantly lowered, although the ATP concentration was diminished and, conversely, the AMP and inorganic P ones were elevated; b. Whereas lindane lowered the glucose effect on the mitochondrial redox potential, it had no influence on the increasing of the ATP/ADP ratio by glucose, or on the antiketogenic effect of this sugar; c. In the animals fed the lindane-contaminated diet, muscular exercise increased the liver betaHOB/AcAc and NADPH/NADP ratios, while the lac/pyr and ATP/ADP ratios were unaltered. But blood pyruvate was increased. The following interpretation has been given. Lindane ingestion inhibits liver mitochondrial activity and increases ketogenesis. The glucose treatment results in a poor glucose utilization for energy needs in the contaminated animals and the forced muscular exercise shows that gluconeogenesis proceeds at a slower rate than in the controls. It is suggested that an increased demand in NADPH, as resulting from the induction of the microsomal enzymes by lindane, is one of the mechanisms by which the pesticide inhibits the activity of the tricarboxylic acid cycle.


Drusen of the optic nerve head.

Drusen of the optic nerve head. Fifty-two eyes with drusen of the optic nerve head from the autopsy and surgical files of the Eye Pathology Laboratory of the Wilmer Institute from January 1967 through December 1975 were reviewed, and histochemical and electron microscopic studies performed. Histopathologic associations with the drusen included 19 eyes with associated optic nerve atrophy, 16 eyes with peripapillary crowding, one case involving prominent dilated vessels pushed aside by the drusen, and one eye with central retinal vein occlusion. Drusen appear to be composed predominantly of a mucoprotein matrix with significant quantities of acid mucopolysaccharides along with small quantities of ribonucleic acid and, occasionally, iron. The literature is reviewed and various correlations and pathogenetic theories are discussed.


Soft lens therapy for recurrent erosion syndrome. Thirty-three eyes of 25 patients with previously unrelieved recurrent erosion syndrome were treated with soft contact lenses. Marked improvement was obtained in 16 cases, moderate in 6, and 11 patients showed no improvement. Patients with visible anterior membrane dystrophies and spontaneous recurrent erosion syndrome responded best. Patients with posttraumatic recurrent erosion syndrome and with recurrent erosion syndrome and with recurrent erosion syndrome associated with aphakic corneal edema did poorly. In addition to having less frequent episodes of stabbing pain, those patients who were improved also had less minor irritation and some had improved vision.


[Study of a fraction antigenically unrelated to the immunoglobulins obtained by immunoadsorption from anti-hapten or anti-protein rabbit sera (author's transl)]. A fraction not antigenically related to the immunoglobulins was obtained together with the antibodies by specific immunoadsorption from either anti-dinitrophenyl or rabbit anti-human gamma globulin antisera. This fraction was called in both cases FIII. It was isolated from specific antibodies by filtration on Sephadex G200. According to the elution volume, its molecular weight is lower than 13,700 daltons. By using specific FIII antisera, it was shown that the FIII derived from anti-DNP antisera is antigenically identical to the FIII derived from anti-IgG antisera. FIII derived from anti-DNP antisera does not react with the free DNP-hapten. FIII was labelled with 125I and it was allowed to react with specific anti-DNP IgG using the radio-immunoelectrophoresis technique technique in which IgG antibodies were revealed by anti-IgG antiserum. It was shown that FIII is combined to the specific precipitate IgG anti-IgG. This result shows that IgG combines either tto the IgG antibodies or to IgG-anti-IgG complexes and suggests that FIII is adsorbed on the immunoadsorbant and not directly through the antigen but through the antibodies or the antigen-antibody complexes.


Lack of inhibition by mouse alpha-foetoprotein (AFP) of in vitro induced lymphocyte blastogenesis. Mouse amniotic fluids (MAF) were tested for their inhibitory effect on the transformation of lymphocytes by concanavalin A, phytohemagglutinin or lipopolysaccharide of Escherichia coli, and on mixed lymphocyte cultures. Eight of the thirteen samples tested contained inhibitory activity equal or superior to 50 % of the controls. Pure preparations of mousealpha-foetoprotein (AFP) were obtained from these or other MAF-pools by immunoadsorption and/or by affinity chromatography on oestradiol-Sepharose. None of the isolated samples of AFP, whether originating from immunosuppressive MAF-pools or from pools lacking activity, exhibited a significant inhibitory effect on the in vitro transformation of lymphocytes. The suppressive properties of the MAF samples and the non-suppressive activity of samples of pure AFP were not affected by the addition of oestradiol-17 beta or after removal of endogenous oestrogens by exhaustive dialysis against 15 % dioxan solutions.


Comparative immunochemical studies of different antigens of deep mycosis pathogens. The disintegration of cells of Coccidioides immitis, Histoplasma capsulatum, Blastomyces dermatitidis and Candida albicans by ultrasonical treatment, repeated freezing-thawing or mechanical methods permitted to obtain antigens with a high protein content. The treatment of these fungal cells with beta-naphthol, ethanol, glycerine or alkali makes it possible to isolate antigens containing mainly polysaccharides. The studied antigens turned out to be complex compounds. Proteins, glycoproteins, nucleoproteins and small quantities of phosphorus and nucleic acids were revealed in their composition. The protein-rich antigens were most active as inductors of a cellular and humoral immune response. These preparations turned out to be the most active and specific for DH testing. The polysaccharide-rich antigens were highly active in in vitro reactions but possessed a low immunogenecity. Immunoelectrophoretical analysis showed a mosaic character of antigen assembly in the preparations isolated from fungal cells and culture filtrates. In different antigens of C. immitis there have been revealed 4-7, in H. capsulatum 3-8 and in B. dermatitidis 3-6 components. The culture filtrates and "cell-saps" had a more complex antigenic spectrum: coccidioidin 7, histoplasmin 8, blastomycin 6; in C. albicans cell-sap, up to 20 antigens have been revealed. In the antigenic spectrum of C. immitis, H. capsulatum and B. dermatitidis common components prevailed, but in C. albicans no cross-reacting antigens have been revealed. Species- and phase-specific antigens have been found in preparations of H. capsulatum.


The induction of immunologic tolerance with type III pneumococcal polysaccharide cross-linked or coupled to protein. Type III pneumococcal polysaccharide molecules were linked to one another, i. e. cross-linked. The product was essentially insoluble in saline. Large pieces weighing 1-2 mg were inserted into mice. They produced immunologic paralysis or immune tolerance. When placed intraperitoneally in millipore chambers the cross-linked polysaccharide did not have any apparent effect, for the mice were not tolerized. When pneumococcal polysaccharide was linked to a protein, the amount of polysaccharide required to cause tolerance or immunity was essentially the same as the amount of free pure soluble polysaccharide. In addition, Xenopus laevis eggs were injected with a different antigen, bovine gamma globulin, and then with antibody to that antigen. Twelve of the twelve eggs so injected changed shape. Equal numbers of other eggs injected twice with the antigen or twice with the antibody did not change shape.


[Effect of actinomycin D, carminomycin and bleomycin and their joint use with serotonin and zymosan on the functional state of the peritoneal macrophages]. The study of the effect of actinomycin D, karminomycin and bleomycin used alone or in combination with zimozan or serotonine on the functional state of the peritoneal macrophages showed that the use of the antibiotics in mice without tumors induced a decrease mainly in the absorptive capacity of the macrophages not changing significantly their digestive activity. The same antibiotics did not significantly change the initial low values of the functional activity of the cells in mice with transplantable tumors. The use of zimozan after completeness of the treatment course with actinomycin D, karminomycin and bleomycin in mice without tumors had a stimulating effect on the functional activity of the macrophages. Zimozan had a stimulating effect on the functional activity of the cells in mice with transplantable tumors only on its use after completeness of the treatment course with actinomycin D or bleomycin. Serotonine in combination with the above antibiotics had no stimulating effect on the functional activity of macrophages in mice both with and without tumors.


[Immunochemical investigations on the gene expression of horse serum carboxylesterase (author's transl)]. Immunochemical and enzymatic analyses of horse serum carboxylesterase were carried out with respect to the existence of a silent gene. Sera with positive phenotypic expression of esterase, both heterozygotes and presumed homozygotes, were compared with:--sera with positive phenotypic expression but genotypically +/O;--sera with a negative phenotypic expression, i. e. genotypically O/O;--sera of natural +/O "hemi-zygotes": mules (donkey lacking the esterase);--positive sera heated at 60 degrees C;--positive sera after specific inhibition of enzymatic activity. Titration by immunocompetition has shown that sera phenotypically negative do not contain an immunologically active "dummy" protein; however, using a sensitive immunotitration (rocket immunoelectrophoresis), traces of active protein could be demonstrated. Sera of +/O genotypes, both horses and mules, contain approximatively half of esterase present in +/+ sera. Heating at 60 degrees destroys the antigenic properties of the esterase almost completely, while, on the contrary, a specific inhibition of enzymatic activity leaves the antigenic properties unaffected.


Killing of intraphagocytic Staphylococcus aureus by dihydrostreptomycin entrapped within liposomes. Liposomes (phospholipid vesicles) may serve as vehicles of antibiotic transport to intracellular sites. Liposomes containing entrapped dihydrostreptomycin enhanced killing of Staphylococcus aureus contained within phagocytic vacuoles.


Relative susceptibility of stream macroinvertebrates to temephos and chlorpyrifos, determined in laboratory continuous-flow systems. Laboratory techniques are described for evaluating the lethal and behavioral impact of pesticides on a range of stream macroinvertebrates under continuous through-flow and simulated stream conditions. The same basic test unit has been used, with slight modifications, to study the reactions of both Simulium larvae and non-target stream invertebrates. On the basis of a standard 1-hr exposure period to different concentrations followed by a 24-hr holding period in a continuous flow of clean water, different test organisms showed wide and consistent differences in tolerance to each of the two insecticides tested. The widest difference between two organisms occurred in the case of the Amphipod, Gammarus pulex (LC90-95, greater than 1 ppm) which was found to be about 5000 x more tolerant to temephos than are nymphs of the mayfly, Baetis rhodani. (LC 90-95, 0.001-0.002 ppm) The widest difference in the reactions of any one species is shown on the part of Gammarus which is about 100 times more susceptible to chlorpyrifos (LC 90-95, 0.05-0.1 ppm) than to temephos. The susceptibility levels of other indicator species such as Agrion, Hydropsyche, Brachycentrus, Ephemera, etc. are discussed in relation to susceptibility levels of Simulium larvae under the same test conditions, and also in relation to current field dosages of the two insecticides in practical and experimental Simulium control.


On ultrastructure of the trabeculae in the interlamellar membranes of nacre conchiolin of the Nautilus shell. The ultrastructure of the interlamellar and intercrystalline membranes of nacre conchiolin of Nautilus has been studied on shadow-cast preparations and on ultrathin sections after various fixation and/or decalcification procedures. After immersion in osmium tetroxyde, glutaraldehyde, EDTA, ascorbic acid and CPC solutions, the nautiloid pattern of conchiolin structure (sturdy trabeculae, elongated fenestration) was observed in all samples. The representative ultrastructure of the sectioned trabeculae was composed of micro-meshworks. Microfibrils were not found. In contrast with the other substance, chromium sulphate produces a shrinkage of the nautiloid pattern into continuous rug-like membranes. In the sections, a distinct condensation of the micro-meshworks occurs at the periphery of both interlamellar and intercrystalline membranes. These observations are discussed namely in relation to the problem of structural identification of the EDTA soluble fraction of the nacre conchiolin.


Effect of vasotocin on plasma free fatty acid level in the migrating anadromous sea lamprey. The effect of injections of arginine vasotocin (AVT) on plasma free fatty acid (FFA) levels was studied in anadromous sea lampreys collected in the St. John River, New Brunswick, during their upstream spawning migration. Plasma FFA was significantly higher in lampreys injected with a single dose of 1 000 mU vasotocin/kg body weight than in those receiving only the vehicle solution, the difference being the greater at 90 than at 30 min post-injection. The significance of AVT in migration is discussed.


Seasonal changes in serum free fatty acid level in the migratory Canada goose. The changes in the serum level of free fatty acids (FFA) in the migratory Canada goose (Branta canadensis interior) breeding in Ft. Churchill (Manitoba, Canada) and wintering in Swan Lake (Missouri, U.S.A.), were studied during the different periods of its yearly life cycle. The lowest serum FFA level was noted during the spring premigratory phase (early March) at Swan Lake, and the highest during moult (early August) at Ft. Churchill. Serum FFA level during the spring postmigratory period (early May at Ft. Churchill) was significantly higher than that during the spring premigratory period and the breeding period (early at Ft. Churchill). No signigicant difference in FFA levels was noted between the fall premigratory (early September at Ft. Churchill) and the fall postmigratory (mid-October at Swan Lake) periods. The significance of the seasonal variations in serum FFA level is discussed in relation to the cyclic physiological events taking place in the bird.


[Cardiac and vascular reactions provoked by 5-HT in rats]. In normal rat, anaesthetized with pentobarbital, 5-HT, injected or perfused by intravenous route, provokes different types of cardiovascular responses. These variations are due to the activation of three classes of receptors. The first ones explain the fall of the arterial blood pressure: they are located on arterioles which are dilatated. The second one are para- or perivascular nervous receptors: when stimulated, they induce vago-vagal reflexes (Bezold-Jarisch reflexes) suppressed by vagotomy. The third ones are vasoconstrictor and located on arterioles.


Determination of useful barley selections in an improvement program for increased lysine content by larvae of Tenebrio molitor L. Larvae of the yellow mealworm, Tenebrio molitor L., Gembloux strain, race F, were reared for 4 weeks at 27 +/- 0.25 degrees C and 65 +/- 5% relative humidity. They were fed on each of 22 cultivars of barley, at the protein level occurring in harvested seed and at a protein level of 10% of dietary protein. Growth and body composition of the larvae were correlated positively and significantly with the concentrations of basic amino acids in the barleys and negatively and significantly with the concentrations of leucine. The percentage of crude protein in larval tissues can be used as a measure of available lysine in barley cultivars, and gains in fresh weight of larvae as indices of arginine concentrations. Differences were evident between the biological and chemical estimations of these amino acids. Several of the Saskatoon barley selections, derived from crosses with Hiproly, were equal to Hiproly or Risø varieties in the amounts of lysine available to the larvae.


Human thyrotropin and its alpha and beta subunits. A new procedure is described for the isolation of human thyrotropin using ion exchange chromatography and gel filtration only. Thyroid stimulating activity of the final preparation of our human thyrotropin amounted to 0.5 IU/mg by bioassay. The alpha and beta subunit of the hormone were also obtained by a new procedure. In this method the native hormone was incubated in an acidified 8 M urea solution and the chains were then separated by ion exchange chromatography and gel filtration. The amino-terminal residues of the alpha and beta chains were valine and phenylalanine respectively. The beta chain appears shorter at its carboxy-terminal end by one methionine residue than its bovine counterpart. Cross-contamination of the subunit preparations were measured by radioimmunoassay. The beta chain exhibited a contamination of about 3 percent of the alpha subunit by weight. The alpha subunit is contaminated by about one percent of the beta chain by weight.


Isolation and characterization of the three light chains from carp white muscle myosin. Myosin from carp white muscle contains two mol of "DTNB" light chain (mol. wt 17 500 daltons) and two mol of "alkali" light chains (mol. wt 25 000 and 16 400 daltons). The three light chains have been isolated in pure state and characterized by electrofocusing, ultraviolet absorption, amino acid analysis and tryptic peptide mapping. Our results show a great homology between the two carp alkali light chains whereas LC2 seems chemically more different. But the homology of LC1 and LC3 is not so extensive as in the case of the higher vertebrate myosin.


[Comparison of the effect of acetone and isopropanol on lipid metabolism in rats]. Isopropanol and acetone administered to rats in conditions leading to a similar blood acetone level differ markedly in their effects on lipid metabolism. Isopropanol administration determines a fatty liver, which is mainly related to a defect in hepatic lipoprotein synthesis. Acetone administration gives only raise to a slight increase in the liver triacylglycerol level. It does not alter the [1-14C] palmitate, [1-14C] glycerol or [U-14C] leucine incorporation into blood lipoproteins. Acetone does thus not appear to play a preminent role in the isopropanol induced fatty liver which seems to be related mainly to a direct action of the alcohol itself.


Interferon inhibits C-type virus at a posttranscriptional, prerelease step. The mode of action of interferon in JLSV 5-cells, chronically infected with Rauscher murine leukemia virus (MLV), was studied by examining the fate of preexisting labelled viral RNA in interferon-treated cells and by determining the infectivity/physical particle ratio of cell-associated and extracellular virus. Interferon added together with 3H-uridine inhibited the production of labelled virus particles even when it was only allowed to act after all viral RNA synthesis had been stopped by actinomycin D. This indicated that the interferon-induced antiviral state primarily functions at a posttranscriptional step. When interferon was given after a 3H-uridine pulse label and arrest of label incorporation by glucosamine and unlabelled uridine, it prevented a portion of the preexisting radioactive RNA from occurring in extracellular particles. However, part of the labelled viral RNA had reached a stage beyond which interferon could not prevent it from occurring in extracellular virus particles. The notion that interferon primarily affects release of fully assembled and enveloped MLV particles may be eliminated: interferon-treatment did not affect the release of particle-bound reverse transcriptase in cells treated with cycloheximide after the antiviral state had been established. It was confirmed that interferon-treated JLSV 5-cells contained an increased number of virus particles associated with the cell membrane. However, these particles were found to have a reduced infectivity compared to those associated with control cells, thus confirming the view that virions produced by interferon-treated cells are defective; perhaps lacking in certain components.


[Ultrastructural organization of the basic substance of human dermis]. The data on ultrastructural organization of the ground substance in the human dermis obtained electron histochemically are represented. Five types of ruthenium positive structures of polysaccharide origin are detected: retinal structure (I), amorfous substance (II), membranes of collagen fibrils (III) and elastic fibres (V), fine ruthenium positive streakness of collagen fibrils (IV). These structures, except fine streakness, form a united polysaccharide system of the dermis participating in maintenance of structural-functional integrity of the connective tissue (collagen-elastic) carcass of the dermis. Two mechanisms, interconnected and oppositely directed, perform this function: the buffer mechanism preventing the connective tissue fibers and collagen fibrils to approach each other, and the binding mechanism preventing the fibrils and fibers to dissociate. The reticular structure performs mainly this function at the level of fibers, and the amorphous substance does it at the level of fibrils.


Ocular involvement in Whipple's disease: light and electron microscopic observations. A 52-year-old man had a prolonged history of nondeforming migratory polyarthritis and a short episode of pericarditis preceding the onset of bilateral vitreitis and retinitis. The clinical course was characterized by progressive deterioration of vision, increasing lethargy, and dementia, leading to coma and death from pneumonia (21 months later). No intestinal manifestations were recorded. Both eyes, which were removed postmortem, disclosed numerous PAS-positive macrophages throughout the inner retina and vitreous. Electron microscopic studies of the macrophages displayed intracytoplasmic, degenerating, rod-shaped bacteria and membranous structures identical to those seen in the intestine, brain, heart, and other tissues of patients with Whipple's disease. Clinicians should include Whipple's disease, and reticulum cell sarcoma, in the differential diagnosis of patients with bilateral retinitis and vitreitis, especially if these disorders are associated with CNS manifestations.


The significance of raised maternal plasma alpha-fetoprotein in twin pregnancy. A hundred twin pregnancies were examined before 27 weeks gestation. The level of maternal plasma alpha-fetoprotein (AFP) was correlated with outcome of the pregnancy. Forty per cent of all women had elevated plasma AFP levels. At birth the combined infant weights were greater in the women with elevated AFP; the gestational age at delivery was unrelated to the level of AFP in early pregnancy.


Neotetrazolium staining of three avian Mycoplasma serotypes for microagglutination test. Mycoplasma meleagridis, M. gallisepticum, and M. synoviae antigens were stained with neotetrazolium chloride for use in the microagglutination test. Only the stained antigens of M. gallisepticum were acceptable in color intensity and test readability.


Amniotic fluid alpha-fetoprotein levels and the prenatal diagnosis of neural tube defects: a collaborative study of 2180 pregnancies in the Netherlands. Amniotic fluid alpha-fetoprotein (AFP) concentrations were measured in 2180 patients at risk of fetal abnormality because of previous history, family history, advanced maternal age, suspected fetal growth retardation and hydramnios. In 12 patients investigated before 20 weeks gestation, pregnancy was terminated because of a raised amniotic fluid AFP-level: 11 fetuses had neural tube defects (NTDs) and one had a congenital nephrosis. There were no false negative results in the 1927 patients tested before 20 weeks and with a pregnancy of known outcome. In patients tested after 20 weeks, the amniotic fluid AFP concentration was raised in 20 cases of anencephaly, in 9 fetuses with severe congenital malformations without NTD and in one apparently normal fetus. Of 428 patients with a previous offspring who had a NTD, only 8(1.9 per cent) again had a fetus with a NTD.


Discrimination of DNA polymerase and RNase H activities in reverse transcriptase of avian myeloblastosis virus. The active sites in reverse transcriptase of avian myeloblastosis virus have been selectively modified by various chemical reagents. The DNA polymerase activity is very sensitive to hydrophobic sulfhydryl reagents such as 5,5'-dithiobis(2-nitrobenzoic acid) and p-hydroxymercuribenzoate but resistant to sulfhydryl reagents with hydrophilic properties. The RNase H activity, on the other hand, is resistant to both hydrophobic and hydrophilic sulfhydryl reagents, indicating the absence of cysteinyl residues essential for RNase H activity. N-Ethylmaleimide (NEM), an amino and sulfhydryl group specific reagent, inactivates both DNA polymerase and RNase H, the later activity being fourfold more stable. Polynucleotides, but not nucleotide triphosphates, protect the two enzymatic activites of reverse transcriptase against NEM. Since pretreatment of the enzyme with 5,5' -dithiobis(2-nitrobenzoic acid) does not prevent N-ethylmaleimide from reacting with a residue necessary for DNA polymerase activity, two different reactive groups are probably involved with this enzymatic activity. The pH profile of reverse transcriptase inhibition by N-ethylmaleimide also suggests the involvement of two reactive groups essential for the DNA polymerase activity with apparent pKas of 5.5 and 6.5. Only one reactive group with a pKa of 7.5 is found associated with the RNase H activity.


Hydrogen-deuterium substitution and solvent effects on the nitrogen-15 nuclear magnetic resonance of gramicidin S: evaluation of secondary structure. Complete assignments of nitrogen-15 resonances of gramicidin S have been made in dimethyl sulfoxide, trifluoroethanol, and in a solvent mixture of dimethyl sulfoxide (50%) and methanol (50%). The assignments are achieved by utilizing the secondary structure of gramicidin S, by comparing the nitrogen-15 spectrum of gramicidin S with that of di-N-methylphenylalanine-gramicidin S and by taking into account the distinguishable value of nitrogen-15 chemical shift for valine in model compounds. Deuterium substitution for labile peptide protons was performed to delineate solvent shielded and deshielded peptide nitrogens and to substantiate further the signal assignments. The solvent titration on going from dimethyl sulfoxide to trifluoroethanol was also performed and shown to have a large deshielding effect on the peptide nitrogen whose corresponding peptide carbonyl, within the peptide moiety, was accessible to the trifluroethanol solvent.


Comparison of the inhibition of thrombin by three plasma protease inhibitors. Human alpha-thrombin is inhibited by the circulating protease inhibitors alpha1-antitrypsin, antithrombin III, and alpha2-macroglobulin. Kinetic analyses of the inhibitor thrombin interactions were carried out utilizing either fibrinogen or the synthetic substrate Bz-Phe-Val-Arg-p-nitroanilide as substrates to determine residual thrombin activity. These studies demonstrated that the inhibition of thrombin by alpha1-antitrypsin, antithrombin III, and alpha2-macroglobulin followed second-order kinetics. The rate constants for the inhibition of thrombin by alpha1-antitrypsin, antithrombin III, and alpha2-macroglobulin are 6.51 +/- 0.38 x 10(3), 3.36 +/- 0.34 x 10(5), and 2.93 +/- 0.02 x 10(4) M-1 min-1, respectively. Comparison of the second-order rate constants and the normal plasma levels of the three inhibitors demonstrates that, under the in vitro conditions utilized, antithrombin III is five times and alpha2-macroglobulin is one-third as effective as alpha1-antitrypsin in the inhibition of thrombin.


[Mediator liberation at the myoneural junctions of the frog sartorius muscle when axoplasmic transport has been disrupted with colchicine]. Colchicine application to the nerve innervating the frog sartorius muscle leads to reduction of the differences in functional condition of the myoneural synapses of this muscle by the signs of quantum content of the end plate potential (EPP) and by the changes of the EPP amplitudes in high frequency stimulation. Along with this there occurs a rarefaction of the frequency of miniature potential of the end plate, and reduction of its amplitude. Apparently, the latter is caused by reduction of effective resistance of the postsynaptic membrane. Changes of the induced and spontaneous secretion of the mediator are connected with disturbances of the axoplasmic transport just in the somatic nerve fibers.


[Alpha-fetoprotein synthesis during liver regeneration in adult mice of different strains]. Alpha-fetoprotein (AFP) concentration was estimated by radial immunodiffusion in agar technique in the sera of adult mice of 12 inbred strains and F1 hybrids (SWR X B10D2 and B10D2 X SWR) during liver regeneration after CCl4 poisoning. Statistically significant difference in the AFP concentration was found in the male and female sera in 6 of 10 mouse strains studied. The following inter-strain differences were also revealed: the AFP mean levels in the sera of C57BL/6 and B10D2 strains were found to be significantly lower than the corresponding levels in the sera of the majority of the strains tested. F1 hybrids showed to occupy an intermediate position by the AFP content between the parental strains. Small, but statistically significant, differences were revealed between the groups of male F1 hybrids from the direct reciprocal crossings. It is suggested that the control of induction of the AFP synthesis during the liver regeneration in adult mice was realized on the polygene basis.


[DNA synthesis in rat liver following administration of antihepatocytotoxic serum]. Experiments were conducted on adult female rats. The autoradiographic method was applied to the study of thymidine-3H incorporation into the parenchymatous and reticulo-endothelial cells of the liver under conditions of using low doses (0.06 microgram of protein per 100 of body weight) of antihepatocytotoxic serum (AHCS), gamma-globulin isolated from it (gammaAHCS) and gamma-globulin fraction of normal rabbit serum (gammaNRS) to intact animals and rats with carbon tetrachloride affection of the liver. The labelled nuclei index of both the parenchyma and the reticuloendothelial cells increased in case of gammaAHCS administration, and, to a lesser extent, of AHCS to intact animals. gammaAHCS used against the background of CCl4 administration intensified the reparative regeneration. The action of gammaAHCS has phasic character--the period of the labeled nuclei elevation was followed by their reduction, replaced by new intensification of the proliferative processes.


Identification and characterization of a brain-specific antigen enriched in neonatal brain. I. Developmental, regional distribution and molecular weight studies. An antiserum raised to the 20--40% ammonium sulfate cut of the soluble proteins extracted from whole neonatal rat brain has been absorbed with liver affinity columns. The absorbed antiserum recognizes a high molecular weight antigen(s) that is (i) restricted to nervous tissue, (ii) enriched in neonatal rat brain, and (iii) present from the 17th day of gestation throughout adult life. The name given to this brain-specific antigen(s) is NABSA, short for 'neonatal-associated brain-specific antigen'. NABSA is immunologically unrelated to S-100 or 14-3-2. NABSA levels in whole rat brain increased soon after birth. The specific activity of NABSA at 2--3 days postpartum reached 3--5-fold higher levels than are found in adult brain. NABSA declined to adult levels by the 10th postnatal day, well before CNS maturation was complete. Regional variations in NABSA levels were found in the rat neonate; highest activities occurred in neonatal cerebellum. Similar variations were observed in adult brain.


Electron microscope autoradiographic evidence for specific transneuronal transport in the mouse accessory olfactory bulb. The distribution of radioactive material was examined autoradiographically 8 h after application of [3H] proline to the vomeronasal organ in mice. Labelled material was transported along the axons of the vomeronasal nerves to their terminals in the glomerular layer of the accessory olfactory bulb (AOB). A lesser but consistent amount of radioactivity was found in the external plexiform layer (EPL) of the AOB. Electron microscopic autoradiography was used to determine which of the components of the EPL contained this labelled material. The method of proportional grain counts showed that the highest concentration of silver grains lay over the mitral cell dendrites, which are the elements immediately postsynaptic to the vomeronasal nerve axons. However, a fairly high proportion of grains also lay over the peripheral processes of granule cells. By application of a method of 'crossfire analysis' (which is explained in detail) it was possible to show that the observed grain distribution is best explained by the assumption that the radioactive material is confined to mitral cells, and the labelling over granule cell processes is due to crossfire from these sources. Im one animal at 5 days after [3H]proline administration label was found to have extended from mitral cells to granule cells, suggesting that the transsynaptically transported radioactive material, which was confined to the mitral cells at 8 h, may have become further redistributed at longer survivals. In a control experiment, [3H]proline was applied directly to the surface of the AOB. This gave rise to a completely different distribution of radioactivity in the EPL: radioactive material was present in all tissue components.