[Changes in antigenic determinant H3 in type A influenza virus]. The analysis of immune and hyperimmune rabbit sera to influenza A/USSR/0249/75. A/Victoria/3/75, and A/Tokyo/1/75 viruses showed the H3 antigenic determinant common for hemagglutinins of influenza viruses of the Hong Kong variety to be poorly manifest in viruses isolated in 1975 in contrast to type A viruses isolated in previous years. Antibody to the H3 antigen is usually absent in immune sera and appears only in hyperimmune sera after reimmunization of the animals. The quantitative decrease of the H3 antigen in viruses isolated in 1975 was also clearly detected in tests of adsorption of anti-H3 monoreceptor sera.


[Electrophoretic mobility of polypeptides in different strains on human influenza  A and B viruses]. The method of electrophoresis in a single polyacrylamide gel plate was used for comparative study on virion polypeptides mobility in human influenza A and B virus strains. Molecular masses of individual polypeptides and their portion in the virion were determined. No variations in the migration speed of nucleoprotein (NP) and membrane protein (MP) were found in strains belonging to the same genus, but there were differences in the migration speed of these proteins in the genus A and genus B. Significant differences in migration and the content of glycoproteins, particularly of the heavy chain of hemagglutinin (HAI) in the genus A strain having different subtypes of hemagglutinin, including strains A/WS/33 (HONI), A/FM/1/47 (HINI), A/Sing/1/57 (H2N2), A/Port Chalmers/73 (H3N2), as well as in the genus B in strains B/Lee/40, B/Yamagata/1/73, and B/Johanesbourg/56. No differences in the mobility of glycoproteins in strains similar in the antigenic specificity of hemagglutinin and neuraminidase were found. On the basis of the comparative analysis of virion polypeptide electrophoregrams, three new strains on influenza A virus isolated in December, 1977, were identified as belonging to the species A (HINI).


Pesticide (hexachlorocyclohexane) inhibition of growth and nitrogen fixation in blue-green algae Anabaenopsis raciborskii and Anabaena aphanizomenoides. The effects of the pesticide hexachlorocyclohexane (HCH) on the nitrogen fixing blue-green algae Anabaenopsis raciborskii and Anabaena aphanizomenoides commonly found as blooms in fish ponds were studied. These algae were very sensitive to HCH, and a distinct decrease in growth rate was observed on prolonged incubation. Lower concentrations (10 microgram/ml) were algistatic and higher concentrations (60 microgram/ml) were algicidal. The inhibition of nitrogen fixation indicated that the presence of HCH might affect overall nitrogen economy of inland waters.


[Report of a study tour in India (author's transl)]. The informant gives an account of the situation of tuberculosis in India (Epidemiology, Tuberculosis Control). Incidence, prevalence and mortality rates of tuberculosis are still high. The results of antituberculosis therapy are in general rather poor. There is existing still a serious problem of chronic tuberculosis. Drug resistant mycobacteria are often found. Nonspecific pulmonary diseases are frequent in spite of a low frequency of smoking.


[Detection of common antigenic determinants in the envelopes of mammalian type D (Mason--Pfizer) and type C virions by using the virion precipitation reaction]. Immune sera to Mason-Pfizer virus (M-PMV) are capable of precipitating type C viruses of mice (NIH-MuLV, G-MuLV, and to a lower degree R-MuLV), cats  (FeLV, RD-114), and monkeys (SSV-1, GALV, BEV). The immune sera to RD-114, BEV, SSV-1 and GALV viruses can precipitate M-PMV virions. The adsorption analysis suggests that cross-reactions between M-PMV, RD-114, NIH-MuLV, and G-MuLV viruses depend on common virus-specific antigenic determinants in the envelopes of the virions. The relationship between M-PMV and type C viruses decreases in the series RD-114--NIG-MuLV--G-MuLV. Type C viruses of mammals have common envelope antigenic determinants lacking in M-PMV. No common antigenic determinants with M-PMV have been found in the envelope of virions of bovine leukemia virus, type B virus of mice (MMTV), or type C avian virus (Pr-RSV).


[Internistic cancer therapy--introduction to the theme]. The importance of internal medicine for the care of patients with carcinoma increases. This is particularly connected with the rapid development of the antineoplastic drug therapy. However, these are not the only tasks of the internal specialists. He is responsible for the ascertainment of the diagnosis, the judgment of the anatomical extension of the tumour with clinical means (classification of stages) and the diagnostics and treatment of concomitant diseases as prerequisites of a rational therapeutic planning. In the oncological team the internal specialist not only represents the specifically internal possibilities of treatment, but is in charge of the continuous general medical and psychic care of the patient. The internal specialist plays an important role in the entire coordination of the specialised care. This refers to the documentation of findings in the carcinoma register of the hospital, to planning and evaluation of controlled clinical experiments, special oncologic consulting hours and consulting functions as a competent specialist also outside oncologic centres, which should be established in district hospitals and teaching hospitals as a common task of all participating disciplines. For this a profound specialisation within the field of internal medicine is necessary. A subspecialisation acknowledged by the state would be a stimulus, but it is not compulsorily necessary. This introduction into the topic shall the internal  specialist make clear his role and responsibility in the oncologic care and facilitate the communication with other specialities.


Study on the nerve endings in the pancreas of Rattus rattus refescens and Gallus domsticus as revealed by the cholinesterase technique. An investigation was undertaken to study the nerve endings in the pancreas of Rattus rattus rufescens and Gallus domesticus by cholinesterase technique. Bulb-like nerve ending in the connective tissue (negatively stained) and dot-like nerve ending in the lobular cells were recorded in Rattus. Dot-like nerve ending on the periphery of the lobular cells and bulb like nerve endings connected with nerve bundle, were recorded in Gallus. Nerve endings were AChE-positive in nature in both Rattus and Gallus.


[Late results following palliative vascular embolization in inoperable kidney tumors. (Preliminary report)]. During 18 months in 15 patients with inoperable renal tumours palliative therapeutic vascular embolisations were carried out with the help of an own catheter technique. In contrast to preoperative embolisation (16 patients) in the inoperable cases a non-absorpable polyvinyl alcohol foam substance ("Ivalon") was used, which leads to a secure permanent vascular occlusion. The haematuria occurring in all patients which many times threateningly increased was removed in all cases. There did not appear any essential complications. However, on account of the formation of a symptomatology of renal infarction a clinical after-treatment is necessary. In 12 patients regulary clinical and paraclinical control examinations during an observation period of on an average 11 months were carried out (minimum 6 months, maximum 18 months). 11 of these patients were still alive at the end of the report and had a tolerable or even improved general condition. In angiographical after-controls of 4 patients, and in one case by autopsy, permanent vascular occlusions with formation of necroses of a large area and partly considerable restriction of the tumour kidneys were found. In these cases the therapeutic embolisation led to a provable damage of the tumorous process. An influence of metastases distant from the primary tumour was not observed.


[Various epidemiological principles of the spread of viral hepatitis in the Astrakhan district]. Analysis of many-year changes in the vitral hepatitis incidence in the Astrakhan region showed a significant cyclicity of the epidemic process: its incidence rose 4 times during the period of from 1957 to 1976; four of these elevations were comparatively high (in 1960, 1964, and 1974). Lately the group of children aged from 3 to 7 years has been of the greatest epidemiological significance, but at present greatest incidence of the disease fell on the group of persons aged from 8 to 14 years. This can be connected with gamma-globulin vaccinations given to children aged under 10 years. Seasonal distribution of morbidity in different are groups was about the same and reflected the general automn-winter seasonal prevalence.


[Prevention of complications of smallpox vaccination]. In observance of the rules of vaccination and unfavourable anamnesis in the vaccinated serve as the main causes of postvaccinal complications (30 and 22%, respectively). The great majority of complications occurs at the age of 1 to 3 years (76%). In 94% of cases complications are recorded in primarily vaccinated individuals. Solution of such problems as introduction into wide practice of sparing methods of immunization and special scheme of preliminary preparation to the vaccination of weak children and those with conditioned contraindications is necessary for the purpose of prevention of complications; a possibility of using, along with gamma-globulin, of synthetic preparations, and of subjecting older children to primary vaccination for prevention of complications is discussed.


Influence of chronic doses of mercuric acetate and lead acetate on the number and activity of Gomori-positive glial cells in the mouse brain. Intraperitoneal injections of mercuric acetate or lead acetate in doses of 0.2 mg daily during 14 and 21 days caused statistically significant rise in the number of the Gomori-positive glial cells around the third cerebral ventricle of mice. In addition, the nuclei of the Gomori-positive glial cells markedly increased in volume.


Uptake and degradation of asialo-fetuin by isolated rat hepatocytes. 125I-labelled asialo-fetuin was taken up by isolated rat hepatocytes by a saturable process. Half maximum uptake was seen at about 3 . 10(-8) M asialo-fetuin. Rate of uptake of asialo-fetuin exceeded rate of degradation at all concentrations of asialo-fetuin tested. Degradation of asialo-fetuin, as indicated by release of acid-soluble radioactivity from the cells, was inhibited by NH4Cl and chloroquine. The intracellular distribution of labelled asialo-fetuin was studied by differential and density gradient centrifuging. The distribution curves for radioactivity indicated that asialo-fetuin was present in lysosomes about 1 h after the uptake had started. Chloroquine and ammonium ions seemed to inhibit the uptake of asialo-fetuin into the lysosomes, possibly by interfering with the fusion between phagosomes and lysosomes.


Inhibition of glycoprotein catabolism in vivo and in the perfused rat liver. Leupeptin is a peptide which inhibits several of the lysosomal proteases. When this compound was added in low concentrations to a perfused liver, the degradation of 125I-asialo-fetuin by the liver was dramatically slowed. When 5 mg leupeptin were added to the perfusate 1 h prior to the radioactive glycoprotein, the liver retained from 70 to 90% or the radioisotope 60 min after infusing 125I-asialo-fetuin. However, untreated livers contained less than 20% of the radioactivity at that time. Subcellular fractionation experiments showed that the radioactivity accumulated in the heavy and light mitochondrial fractions (ML) of the homogenate. At 80 min after the glycoprotein was added, almost 40% of the radioactivity was still located with these fractions. Very similar inhibitory effects were seen upon treating rats intravenously with 5 mg of leupeptin 60 min prior to injection of 125I-labelled asialo-fetuin. A 7 fold increase in liver radioactivity was observed 6 hrs after the glycoprotein had been given to the treated animals. Purified human liver cathepsin B digested fetuin to about 3% of total hydrolysis and the major peptide fragment produced had an SDS-electrophoretic mobility equivalent to that of ovalbumin.


A comparison of methods for blocking staining of nucleic acids. Using rat spinal cord as test material 4 methods for blocking staining of nucleic acids with simple basic dyes have been compared. Two of these methods--a perchloric acid procedure for tissue blocks and a Ba(OH)2 technic for sections--block staining of RNA. The other two-immersion of sections in solutions of dichlorodiammine Pt II or in 10% zirconyl chloride-blocked staining of both RNA and DNA. None of these methods hydrolyze DNA sufficiently to produce a positive Feulgen and no loss of protein could be demonstrated. With the three metal containing procedures some metal could be demonstrated in the sections. This deposition was primarily in nucleic acid containing structures and was greatest and most widespread with the zirconyl chloride. The latter method altered the color of all hematoxylin technics to a deep red. The separation of nuclear stains on the basis of their bonding characteristics into three groups (Lillie et al. 1976) was confirmed.


[Essay of proteins after aldehyde treatment in biological objects (author's transl)]. Following fixation with formalin or glutaraldehyde, the protein content of bovine serum albumin, lysozyme, non-disintegrated yeast cells and liver nuclei of mice is determined by means of a brom-phenol blue staining procedure and the widely used Lowry-technique. The bromphenol blue technique permits determinations of soluble as well as particulate proteins or protein mixtures fixed with up to 6% formalin or 2% glutaraldehyde. Recovery rates differ no more than 20% as related to unfixed controls. Using the bromphenol blue method it is not necessary to separate aldehyde or any interfering material by additional steps prior to determination. Factors for correcting protein content of biological material after aldehyde treatment are available. In this way, comparative biochemical as well as cyto- and histochemical investigations of enzymatic activities after aldehyde fixation are possible. Some advantages of the bromphenol blue technique with special reference to the analysis of particulate and/or aldehyde-fixed specimens are discussed.


[The cytospectrophotometrical determination of proteins by the tetrazonium coupling reaction. I. Optimization of the staining method (author's transl)]. The tetrazonium coupling reaction using Fast blue salt B and the sodium salt of 6-hydroxy-naphthalinsulfonic acid-(2) serves as a method for histochemical demonstration of proteins. Fast blue salt B reacts with histidin, tyrosin and in a slighter extent also with tryptophan. It is shown in this paper, that the histochemical tetrazonium reaction carried out under the optimal conditions is a specific staining for proteins and highly reproducible. The maximum of absorption at lambda = 530 nm of the azo dyes generated by the coupling reaction followes the Lambert-Beer's law. Nucleic acids as well as protein-SH-groups do not react with Fast blue salt B under the conditions described. Due to the exact reproducibility and specifity the tetrazonium coupling reactions seems to be a suitable method for at least comparative, microspectrometric protein determination within one species of cells.


[Pulse cytophotometric determination of lipids in cultivated animal cells (author's transl)]. The determination of lipids in cultivated animal cells (aortic smooth muscle cells and L cells) by pulse cytophotometry is described. The cells were stained with phosphene 3 R and benzopyrene caffeine. By standardization of the procedure a reproducible quantitative evaluation was possible.


A neuronal nucleolar trigger mechanism traceable by a routine staining method. In experimental Wilson's disease produced by the intracardiac injection of copper sulphate, it is seen that the glia-neuronal interactions play an important role in the pathogenesis of neural lesions. A "trigger" copper-RNA particle is transferred from the oligodendroglial nucleus to activate the neuronal nucleolus causing a flooding of the neurone with RNA. This trigger particle stains differently with Mason's trichrome, giving a bright red colour. This altered reaction gives an insight into the feedback process for the formation of ribosomal RNA in the neurones. The alteration appears to lead to a block in the repressor activity of the neuronal RNA formation.


Concentric lamellated psammoma-like bodies in the third ventricle of the European mole rat (spalax leucodon, Nordmann 1840). Concentric lamellated bodies are described in the third ventricle of the European mole rat. These bodies show most staining characteristics of true psammoma bodies found in humans but were never calcified in the mole rat.


Vital staining properties of neutral red. Vital staining of cornea and conjunctiva. Vital staining by instillation in the conjunctival sac of 1% neutral red in 54 normal and 153 differently affected eyes was studied by slit lamp examination. Of these, 134 eyes were after-stained by a mixture of 1% tetrazolium and 1/4% alcian blue and 73 eyes were after-stained by a mixture of 1% rose bengal and 1% fluorescein. Microscopy of a further 10 conjunctival scrapings and 40 mucous conjunctival threads disclosed neutral-red-stained inclusion bodies in the cytoplasm of the epithelial cells and the granulocytes. In the slit lamp the cornea and the conjunctiva were seen to be stained only rarely and, if so, poorly by neutral red, both in normal and affected eyes. Better staining was obtained with tetrazolium, and the best with rose bengal. The plica semilunaris, Marx' line, and the mucous thread were stained by neutral red, more intensely by rose bengal, and the least by tetrazolium. The inferior fornix and the tarsus were rarely stained by any of the three dyes. Neutral red seems to offer no diagnostic advantages over the vital stains with which the dye has been compared in the present study. More particularly we found no pathological processes that were stained more intensely by neutral red than by the other vital stains.


Early morphological changes in organ cultured human corneal endothelium. Nineteen human cadaver corneas with few damaged endothelial cells were incubated under tissue culture conditions for time periods ranging from five min to 48 h. Morphological alterations of the endothelial cells were studied in whole wet mounts stained by alizarine red-alkohol-trypane blue and by scanning electron microscopy. Joint meetings of three cells are characteristic for normal corneal endothelium. After 15--60 min of incubation, damaged cells were expelled from the coherent cell sheet by expanding neighbouring cells. Joint meetings of 5--8 expanding cells were formed. After 24 h of incubation, joint meetings of four cells were the dominating morphological abnormality. Morphological changes during reduction of the numbers of cells in joint meetings are described.


The precision of unbiased estimates of numerical density of endothelial cells in donor cornea. The precision of estimates of central corneal endothelial density was studied in 16 human corneas stained by alizarine red and trypane blue. Estimates based on central counts and estimates based on peripheral counts were considered separately. The numerical density was estimated employing an unbiased sampling technique. From central counts estimates with an error of less than five per cent could be obtained. From peripheral counts a maximum precision of mean +/- 12.2 per cent could be obtained. The theoretical maximum precision was calculated by application of a variance component model. The precision of estimates was calculated for 1, 2 and 4 test areas of different sizes. Economy of sampling was evaluated by comparison of the actual precision to the theoretical maximum precision of estimates.


Contaminants in human milk.

Contaminants in human milk. There is a paucity of information regarding excretion of contaminants in human milk, due to experimental difficulties and until recently a general lack of interest. Because of the high fat content of milk and as its acidity is higher than that of plasma, nearly all liposoluble and basic agents consumed by the mother will be excreted in the milk. Distinction must be made between, on the one hand drugs and social toxicants such as smoking and alcohol, whose intake can be stopped or limited during pregnancy and lactation, and ecological toxicants present in a polluted environment to which the mother is exposed. Cases have occurred of heavy prenatal and postnatal intoxication of infants with hexachlorobenzene in Turkey and methylmercury in Iraq due to consumption of fungicide-treated seed wheat by pregnant and lactating mothers. Recent attention has been concentrated on contamination of milk with organochlorine compounds such as DDT and PCB's, that are found in many parts of the world. The heaviest contamination with DDT has been found in Guatemala, resulting in suckling infants consuming many times the Acceptable Daily Intake of this compound proposed by WHO, with unknown future effects.


Transient intestinal lymphangiectasia.

Transient intestinal lymphangiectasia. Intestinal lymphangiectasia may be more protean in clinical manifestations and less rare than earlier suspected. A recent report points out that there are two types of the disorder, one congenital and the other acquired and transitory. A case is reported which fulfills the current clinical, laboratory, radiological and histological criteria for the diagnosis of the disease, and represents the first report in Scandinavia of transient intestinal lymphangiectasis with rapid and complete recovery within a few months after initiation of MCT diet.


Fluorescent labelling of cell membranes and cytoplasmic proteins in living cells. Primary amino groups may be selectively labelled in living cell cytoplasmic components by staining with the covalently binding fluorochrome reagent fluorescamine. The reaction is extremely rapid and occurs at very low reagent concentrations. Cells survive such treatment and gradually remove or metabolize the labelled substances. Nuclei and nucleoli are not labelled, while lamellar cytoplasm, which contains little actual cytoplasm, is well demarcated, thus indicating that the method is useful for studies of cell membrane components. Labelled cell membranes can be prepared for further purification after preliminary external fixation of cell membranes with a supravital polyaldehyde fixative. The use of dimethylsulfoxide as a solvent for fluorescamine allows much longer survival of the cells than the use of acetone as a solvent. In addition, the use of high pH and borate buffer was not necessary to the labelling phenomenon.


[Study of thyroid function using diverse methods of separation of plasma radioiodine compounds in rats]. We compared the fractionations obtained by three different methods of dosage of thyroid hormones and we concluded that trichloracetic PBI is very rapid, (about 15 minutes) excluding the length of calculation of radio activity), is easy to execute, and uses inexpensive materials. Separation by exchanging resins of ions is also very rapid (15 min.), a little more precise, but the cost of operation is much higher. Separation by sephadex G25 is very time consuming (one day), and much more meticulous; its cost is equally high.


Drug-induced neutropenia in the Stockholm region 1973-75: frequency and causes. The records of 133 patients, discharged with a diagnosis of "agranulocytosis" (i.e. blood neutrophil levels less than 1.0 x 109/1) during the years 1973-75 in the Stockholm county region, were reviewed. In 45 cases the neutropenia was probably caused by drugs other than cytostatics, giving an annual incidence of drug-induced neutropenia of 0.01%. The most common drugs were thenalidine, sulfonamides and thyreostatics. Only one of the 45 patients died during the neutropenic phase. It is concluded that the pattern of drugs causing neutropenia has changed in Sweden compared with studies from the latter half of the 1960's, and only about 40% of the cases have been reported to the authorities.


Bacterial decomposition of synthetic 14C-labeled lignin and lignin monomer derivatives. Nocardia sp. which was isolated from soil is capable of degrading synthetic lignin and utilizing its monomer derivatives. Decomposition was monitored by measuring the 14CO2 evolved and O2 consumed, when the bacterium was grown on a medium containing specifically 14C-labeled ligning or monomer phenolic compounds as major carbon source. The time course of the 14CO2 release and O2 uptake indicates a significant depolymerization and utilization of lignin by the Nocardia sp.


The effect of certain mannich N-bases, derivatives of isatin beta-thiosemicarbazone, on the replication of vaccinia virus in in vitro studies. Twenty-three Mannich mono- and bis-N-bases, derivatives of isatin beta-thiosemicarbazone (IBT), were synthetized. Eighteen of these were tested for their inhibitory effect on the replication of vaccinia virus (IHD strain) in cultures of chick embryo fibroblasts. All the compounds studied showed antiviral activity, demonstrated by the agar-diffusion method (screening). The concentrations that reduced the number of viral plaques to 50% (PRD50) and the values of the therapeutic index (Th.i.) of the compounds tested were determined by the methyl-cellulose technique. The results obtained permitted analysis of their pharmaceutical effect relative to the chemical structure of the molecule, under the described experimental conditions.


Leucocyte migration inhibition test as an evidence of immunological response to measles virus. III. Statistical analysis. Two-way analysis of variance with replications was used for comparing the size of leucocyte migration areas in the samples of blood of monkeys before and after immunization of animals with measles virus. The areas in control samples were compared with those with antigen. Size of migration areas differed significantly between control samples and those with antigen only after the immunization of animals. Interaction between individual monkeys and presence or absence of antigen was significant only 28 days after the immunization of animals.


Some factors affecting the heterotrophic activity of bacteria in lake. The heterotrophic activity of aquatic bacteria was measured by incorporation of 3H-thymidine, uptake of 14C-sodium acetate and mineralization of 14C-acetate. The temperature of water, oxygen content and concentration of chlorophyll a were also measured in the studied lakes. The obtained results of the studied processes in the surface layer and epilimnion were distinguished by negative correlation coefficients. A distinctive feature is the relation between chlorophyll a concentration in the water bodies and heterotrophic activity of bacteria. This indicates a negative dependence between these processes and phytoplankton activity in the photic zone. In contrast a positive relationship between phytoplankton activity and heterotrophic activity of bacteria was found in the metalimnion and hypolimnion of the studied lakes.


The estimation of extracellular release by phytoplankton and heterotrophic activity of aquatic bacteria. The actual extracellular release of organic matter by algae was determined in water samples in which the heterotrophic activity of bacteria was inhibited by gentamycin. Gentamycin rapidly and efficiently inhibited the activity of aquatic bacteria without affecting phytoplankton metabolism. Aquatic bacteria utilized the products of algal extracellular release. The amount of algal photosynthetic products metabolized by bacteria can be taken as a measure of their heterotrophic activity in waters.


Effect of copper and tri- and hexavalent chromium on the work of an activated sludge. The effect of metals (Cu2+, Cr3+, Cr6+) on the work of an activated sludge grown in crude oil refining or synthetic wastes was examined. The activated sludge method was found to be applicable for the purification of wastes carrying up to 0.8 mgCu2+/1,15mgCr3+/1, or 20mgCr6+/1. Higher concentrations of these metals inhibited the work of the activated sludge which was evident in inferior purification and reduced intensity of respiration of the activated sludge microorganisms.


The effect of cyclohexane derivatives on selection of bacterial groups forming activated sludge microflora. The effect of cyclohexanol, cyclohekxanon and cyclohexylamine on the selection of bacteria in a model population composed of bacteria isolated from activated sludge was examined. The initial population consisted of both gram-positive and gram-negative bacteria. The latter, which accounted for 90-97% of the population, belonged mainly to three Pseudomonas groups and the Enterobacteriaceae, Vibrio-Aeromonas, Achromobacter-Alcaligenes and Flavobacterium groups. Seven day growth in medium containing cyclohexane derivatives caused pronounced qualitative changes in the population. The compounds favored the development of bacteria of the genus Pseudomonas and inhibited the growth of all other gram-negative bacteria. The direction of selection was independent of the type of cyclohexane derivative.


Studies on the sterilization of pharmaceutical base materials with ionizing radiation and ethylene oxide. The use of ionizing radiation and ethylene oxide for the sterilization of pharmaceutical base materials of animal origin, used to produce organopreparations, was studied. The materials included liver extract, pancreas extract, dried thyroid and intrinsic factor. The effective sterilizing doses for the examined materials and dependence between effective ionizing radiation dose and primary contamination were determined.


Genetic transformation in Rhizobium trifolii. Markers controlling the synthesis of amino acids and organic bases as well as streptomycin resistance and sensitivity to acriflavine were transformed in Rhizobium trifolii. The results indicate that the str marker was transformed independently of leu, his, ade and trp markers. Co-transformation of leu and utra markers ranged from 3 to 7%, whereas that of thi and acr was 10%.


Isolation and preliminary characterization of R6K plasmid deletion mutants. Several deletion mutants of R6K have been isolated by mutagen treatment of bacterial host carrying wild type of the plasmid and search for clones that lost ampicillin or streptomycin resistance. The molecular weight of the mutants as estimated by agarose gel electrophoresis was 15 times 10(6) minus 23 times 10(6) compared to 26 times 10(6) for the parental plasmid. The mutants were characterized in respect of the level of resistance to ampicillin and frequency of conjugational transfer. Some of the mutants were found to differ in Eco RI digestion pattern from the wild type.


Lipopolysaccharides of Shigella flexneri 6 as phage receptors. Seven bacteriophages specifically lytic for representative strains of Shigella flexneri serotype 6 were studied. Cross hemagglutination experiments and chemical analysis of isolated lipopolysaccharides showed that strains 975 and 323 are smooth and strains 288, 551 and 488--rough. Lipopolysaccharides isolated from the smooth strains inactivated phages F10 and "a" whereas those from the rough strains demonstrated poor inactivation of specific bacteriophages. The lipopolysaccharides of both the smooth and rough strains were found to be phage "a" receptors. The results indicate that the varied sensitivity of S. flexneri 6 depends on the roughness of the strains.


Studies on the action of myelin basic protein (MBP) in rat brain. The specificity of I125-MBP uptake and subcellular distribution on a discontinuous sucrose density gradient, compared to those of histone H 4 and cytochrome c, showed a high affinity of MBP for mitochondria and heavy synaptosomes, and of histone H 4 for lighter synaptosomes. One heavier synaptosomal subpopulation was almost equally labelled by both proteins. Cytochrome c showed only a low uptake into particular material. Receptor interaction studies of MBP with H3-labelled 5-hydroxytryptamine and naloxone gave negative results.


In vivo incorporation of 32P into myelin basic protein from normal and quaking mice. Myelin basic protein in normal mice is phosphorylated. Since phosphorylation can decrease the net positive charge of the myelin basic protein, this could affect molecular interactions between this protein and other myelin components. In this study 32P incorporation into small and large components of the myelin basic protein was studied in immature and young adult mice and also in Quaking mutants which have a severe myelin deficit. We found a short half-life of 32P in myelin basic protein. The 32P specific activity of myelin basic protein was higher in immature and Quaking mice than in young adult animals. Of the 32P-labeled basic proteins of control and Quaking mice, the small component had a slightly higher specific activity than the large component. Although the small basic protein is quantitatively decreased in Quaking mice, the ratio of specific activity of small to large basic protein is similar in control and Quaking animals. Since Quaking and immature mice have many uncompacted myelin lamellae, these preliminary results suggest that phosphorylation and dephosphorylation could be involved in compaction mechanisms.


Biosynthesis of myelin and neurotoxic factors in the serum of multiple sclerosis patients. The in vitro synthesis of myelin proteins has been studied by measuring the incorporation of [3H] lysine in developing rat brain slices. This incorporation system has been used to assay potentially gliotoxic and myelinolytic agents. A reduced incorporation of the labelled amino acid into myelin proteins occurs in the presence of anti-myelin anti-serum and anti-basic protein anti-serum. Diphtheria toxin has been found to inhibit the synthesis of myelin basic and proteolipid protein in the white matter slices of developing rats. Recent experiments with serum samples from multiple sclerosis patients in exacerbation suggest the presence of a factor which interferes with the synthesis of myelin in white matter slices.


Conformation of myelin basic protein and its role in myelin formation. High resolution 13C and 1H NMR spectra of myelin basic protein over a range of pH and concentrations indicate that intramolecular folding of the polypeptide chain occurs in aqueous solution in the region of residues 85 to 116. At pH 4 in D2O solution, the 13C resonances due to nonprotonated carbons of phenylalanine and tryptophan are broadened and chemically shifted compared to the same resonances when the protein is dissolved in 6M guanidinium hydrochloride. These residues occur in the region of the polypeptide chain in which the intramolecular folding may occur. As the pH is raised and the positive charge on the protein reduced from 28 to 18, intermolecular aggregation occurs, which appears to involve these same folded regions. Data on T1 (longitudinal relaxation times) of protons indicate also that amino-acid sidechains vary considerably in their motional freedoms. The concentration dependence of the proton NMR spectra provides further information on association of protein monomers. The region of the protein involved in folding, polymerization and substrate specificities is conservative in various species and we can surmise that it may have a specialized role in protein-lipid interactions in the myelin membrane. We suggest that the protein forms dimers across the cytoplasmic apposition during the formation of myelin. Estimates of the repulsive energies of interaction between approaching membranes suggest that some special mechanism of this kind is required to overcome the repulsive forces due to breakdown of water structure and electrostatic interaction.


Cross-linking of lipid bilayers by central nervous system myelin basic protein: aggregation of free and vesicle-bound protein. Central nervous system myelin basic protein binds to the zwitterionic lipid, egg diacylphosphatidylcholine, over a wide range of pH and ionic strength. Lipid vesicles containing the protein have been observed to increase in size and to aggregate. The size increase is most marked at very low ionic strengths whereas aggregation is evident at ionic strengths from 0.001 to 0.35. The pH and ionic-strength dependence of this aggregation closely follows that of the self-association of the protein, suggesting that vesicle association is mediated by binding between polypeptides attached to different vesicles. Basic protein is monomeric at low pH but above pH 6 self-associates yielding primarily small oligomers (probably dimers) and minor amounts of higher species. It is envisaged that each protein molecule possesses two distinct binding sites, one capable of association with lipid bilayers and the second with another protein molecule. Basic protein is found predominantly on the intracellular surface of the myelin membrane. Given the ability of the protein to act as a bridge between lipid bilayer vesicles in vitro it is proposed that it may perform a similar function in vivo, serving to cross-link the inner surfaces of the oligodendroglial cell membrane. This protein function could lead to formation of the long cellular processes which encircle the nerve cell axon and could assist in stabilizing the highly ordered myelin structure which results.


Molecular organisation in central nerve myelin. Pertinent data from the literature and in press is summarised and used to construct a model for the molecular arrangement of lipid and protein in the lamellae of compact central nerve myelin. For the lipid phase of myelin the available data is best interpreted in terms of a bilayer arrangement while physical studies suggest that the lipids are in an intermediate fluid state maintained by the presence of cholesterol and water in the system. Lipids will interact to maintain this condition. The proteins of myelin differ in their membrane locations. The high molecular weight proteins are considered to be intrinsic components with at least part of their polypeptide chains in the lipid phase. The proteolipid protein is also intrinsic and may be completely buried in the lipid phase. The basic protein of myelin is an extrinsic component and must be localised at the surface of the lipid phase at either the external or cytoplasmic face of the lamellae. Present results suggest an elusive location at the cytoplasmic apposition region. The lipid-interacting properties of the basic protein are segregated on the polypeptide chain of the molecule and this may be important for the possible role of the basic protein in bridging adjacent lamellae at the cytoplasmic apposition. It is speculated that association of the proteolipid protein with the basic protein in a 1:1 molar ratio would form an effective lipid-complexing nucleus in the lipid rich myelin lamellae but experimental data to support this idea is lacking at present.


Autoimmunity in multiple slcerosis: do we have an experimental model? Experimental autoimmunity of the CNS has been well characterized--the antigen has been identified, effector cell specificity has been defined, and the relationship between cellular sensitization and antibody production has been partially clarified. In the guinea pig, experimental allergic encephalomyelitis (EAE) is induced by one injection of myelin basic protein in complete Freund's adjuvant (BP/CFA). If BP/CFA is preceded by repeated injections of basic protein in incomplete Freund's adjuvant (BP/IFA), EAE is not induced; the guinea pigs survive and ultimately produce antibody. Induction and prevention of EAE as well as antibody induction by this schedule are dependent on the presence of the intact encephalitogenic (T-cell) site in the polypeptide used for sensitization and preimmunization. In contrast, B cell sites (those peptide sequences which bind antibody) are independent of the T-cell site. At least 5 specific antigenic regions (B-cell sites) have been demonstrated in the BP molecule. High mycobacteria levels bypass the specificity requirement of helper T-cells but cannot bypass the specificity requirement of effector T-cells. In spite of the sophisticated immunologic techniques available, our knowledge of humoral and cellular sensitivity in multiple sclerosis (MS) patients is very limited. The experimental demonstration of an analogy between EAE and MS is weak: a) Demonstration of BP-sensitized cells or BP-specific antibodies in peripheral blood of MS patients has not been successful. b) Anti-myelin serum factors reported to be associated with both disease states (experimental autoimmunity and MS) are clearly not identical. Nevertheless, successful treatment of EAE in animals by BP/IFA injections has encouraged consideration of clinical trials to test the therapeutic value of BP injections in MS patients. If successful, the question will be answered: if unsuccessful, the dilemma still remains.


Molecular bases for the difference in the potency of myelin basic protein from different species in Lewis rats. The results of this study define the chemical bases for the difference in the encephalitogenic potency reported for the bovine and guinea pig myelin basic proteins. Studies with synthetic peptides showed that the sequence of peptide S53, H-Ser-Gln-Arg-Ser-Gln-Asp-Glu-Asn-OH, which is native to the guinea pig basic protein, is the minimum amino acid sequence necessary for inducing EAE in Lewis rats. The results of this study further showed that specific sequence modifications rendered the native bovine sequence highly encephalitogenic.


Clearance of myelin basic protein from blood of normal and EAE rabbits. The rate of clearance of porcine myelin basic protein (MBP) from plasma of rabbits was determined following intravenous injection of 20 mg MBP. The MBP level in the plasma was measured by a 2-site immunoradiometric assay with specific antibody to guinea pig MBP produced in rabbits. Plasma MBP-antibody levels were determined by competitive binding radioimmune assay (RIA). Unsensitized and those sensitized with complete Freund's adjuvant (CFA), with porcine MBP in CFA, and with whole porcine spinal cord in CFA were studied. Unsensitized and CFA sensitized rabbits exhibited maximum MBP levels in the plasma within two minutes after injection with rapid decrease to undetectable levels in one hour. Thirty-nine of the unsensitized (control) rabbits exhibited normal, rapid clearance and no subsequent physical signs of EAE while one of the control rabbits exhibited a slightly retarded clearance rate. Histologic examination of autopsy tissues from the control group revealed that five rabbits showed lesions which could be attributed to Encephalitozoan cuniculi or Toxoplasma and one rabbit autopsied 65 days after clearance had minimal EAE lesions. Rabbits sensitized with MBP exhibited a retarded rate of clearance at the acute stage of EAE and following recovery. Rabbits sensitized with whole spinal cord in CFA also exhibited a retarded rate of MBP clearance. Anti (MBP) antibodies were detected in the plasma of all rabbits which exhibited a retarded rate of MBP clearance. Significant rates of retardation were not detected until approximately three weeks after sensitization with CFA-MBP or CFA-spinal cord. While MBP antibody levels in most animals were not detected by the immunodiffusion technique, antibodies were demonstrated by RIA. The 20 mg MBP given intravenously is probably in great antigen excess and conducive to the formation of soluble MBP-anti (MBP) complexes in the blood.


Baisc protein hydrolysis in lymphocytes of Lewis rats with experimental allergic encephalomyelitis. Lymphocytes from lymph nodes of Lewis rats with acute experimental allergic encephalomyelitis (EAE) contain high amounts of acid and neutral proteinases which hydrolyze myelin basic protein. The activity at neutral pH is also expressed by whole lymphocytes in isotonic medium, with about 50% more activity released by homogenization. Neutral proteinase activity in lymphocytes increases with the onset of acute EAE while the activity of those from Freund's adjuvant-injected controls increases somewhat later. The total neutral proteinase activity appears to be membrane-bound, most likely in the lysosomes, but half the total was associated with the nuclear fraction. The basic protein proteinase was compared with an enzyme described earlier, especially active toward polylysine, and some differences were noted. It appears that two enzymes may be present in lymphocytes which hydrolyze basic protein at a neutral pH. An increase in neutral proteinase activity was observed in some, but not all, lymphocyte preparations from patients in various stages of multiple sclerosis. The finding that whole activated lymphocytes are capable of hydrolyzing basic protein suggests that these cells which are believed to be precursors of mononuclear cells migrating into the central nervous system may be active agents in the early stages of myelin dissolution in experimental allergic encephalomyelitis. At present, such a mechanism is only theoretical, and the possibility that activated lymphocytes may be a factor in demyelination in multiple sclerosis is even more speculative.


Neutral proteinases secreted by macrophages degrade basic protein: a possible mechanism of inflammatory demyelination. In the inflammatory demyelinating diseases, such as multiple sclerosis, Landry-Guillain-Barré syndrome and experimental allergic encephalomyelitis, demyelination occurs in the vicinity of infiltrating mononuclear cells. Although the histopathology is characteristic of each disease, the general observation that myelin destruction in inflammatory lesions begins prior to phagocytosis suggests a common mechanism for myelinolysis in these diseases. Recent studies show that stimulated macrophages secrete several neutral proteinases, including plasminogen (Plg) activator. We have tested the possibility that these proteinases could, directly or indirectly, initiate myelin destruction. Isolated brain myelin was incubated with supernatant media from cultures of stimulated mouse peritoneal macrophages in the presence and absence of Plg. Cell supernatants alone caused some degradation of basic protein (BP) in myelin. The amount degraded was considerably enhanced in the presence of Plg. The other myelin proteins remained essentially intact. While the Plg-independent proteolytic activity in the supernatants was abolished by EDTA, known to inhibit the neutral proteinases, the Plg-dependent hydrolysis was inhibited by p-nitrophenylguanidinobenzoate, an inhibitor of Plg activator and plasmin. These results suggested that the Plg activator secreted by the macrophages generated plasmin, which selectively degraded BP. This interpretation was confirmed by the observation that urokinase, a Plg activator, plus Plg was effective in degrading BP in myelin. We propose that the action of neutral proteinases released by stimulated macrophages, and its amplification by the Plg-plasmin system, may play a significant role in several inflammatory demyelinating diseases; and that the relative specificity of these reactions for myelin lies in the extreme susceptibility of BP to proteolysis.


Effect of diphtheritic demyelination on axonal transport in the sciatic nerve and subsequent muscle changes in the chicken. Chicken sciatic nerves undergo demyelination following intraneural injection of diphtheria toxin and subsequent atrophy of some muscular cells. Paresis occurs after one week and lasts approximately three weeks; at the height of the lesion C14-leucine was injected into the ventral horn cells of the spinal cord. The axonal transport of fast flowing labelled proteins was followed down the sciatic nerve axons and flow rates at two different times were measured. Muscle cells were stained for succinic dehydrogenase and ATPase; fibre diameters, total protein, and total radioactivity associated with the nerves were also measured. The results showed that the fast flowing labelled proteins accumulated at the demyelination site while the muscle cells supplied by these nerves showed reduction of fibre diameter and evidence of degeneration. Further studies are in progress on slow moving proteins and muscle cells.


Cellular and humoral responses to myelin basic protein in multiple sclerosis: a dichotomy. The macrophage migration inhibition factor (MIF) assay and a counterimmunodiffusion assay were utilized to measure immune responses to human myelin basic protein in 75 patients with multiple sclerosis (MS) and in 120 control subjects. Eight out of ten MS patients in acute exacerbation and one out of seventeen convalescent, but none of chronically ill MS patients gave positive results in the MIF test. Forty-six percent of the patients with negative MIF assays but only 22% of those with positive assays had positive antibody results. In the counterimmunodiffusion assay, myelin basic protein antibody was demonstrated in almost 2/3 of patients during convalescence but it was not present in those whose illness had been stable for 6 months or longer. While no correlation with the stage or duration of the illness was present in other disorders, in MS an inverse correlation with clinical activity and in vitro evidence of cellular sensitization to encephalitogenic basic protein was apparent.


Cerebrospinal fluid myelin basic protein and multiple sclerosis. We have previously reported that myelin basic protein appears in CSF during acute attacks of multiple sclerosis. These studies have been extended to over 700 patients, 91 with multiple sclerosis. The data continues to indicate that myelin basic protein is released into cerebrospinal fluid during acute attacks of multiple sclerosis. We are currently characterizing the basic protein in the CSF.


Proteolytic activity in CSF.

Proteolytic activity in CSF. Proteolytic enzyme activity, present at both acid and neutral pH values, in cerebrospinal fluid, can be measured by a sensitive assay, which monitors the rate of 125I-basic protein breakdown on polyacrylamide gel electrophoresis. CSF cellular neutral proteinase and supernatant acid proteinase are increased in acute multiple sclerosis and in CNS infections.


Isoelectric focusing and isotachophoresis for investigation of CSF and serum proteins in demyelinating and infectious neurological diseases. Isoelectric focusing (IEF) and isotachophoresis (ITP), two methods with excellent separation capacities, have been adapted during recent years for the analysis of CSF proteins. The fractions separated by these techniques can be further studied by e.g. immunological methods. ITP has besides its high separation capacity several valuable advantages: very small samples are needed, unconcentrated CSF can be examined, the analyses are quickly performed and the results are immediately obtained on a recorder. Examinations by thin-layer IEF in a series of about 2,000 patients have afforded much new information about the CSF and serum proteins in many neurological diseases. Different complex CSF protein aberrations have been found in the gammaglobulin range as well as in more anodal positions in MS, infectious neurological diseases and Guillain-Barré syndromes. These aberrations are probably the result of several interacting factors, e.g. the temporal and spatial characteristics of the disease, the release of decomposition products from destroyed tissues, the genetically determined reactivity of the individual and the type of etiological agent.


Studies on myelin proteins in human peripheral nerve. The myelin fraction from human peripheral nerve was prepared. Two basic protein fractions (BF-P2 and PB) were isolated from acid extracts of the myelin fraction and three glycoproteins (BR-PO, PASII and Y protein) were purified from its acid-insoluble residue. In biochemical analysis the human BF-P2 protein (M.W.13,000) showed similar but not identical properties to bovine BF-P2 protein. The PB fraction was suggested to include the encephalitogenic CNS-BP (M.W.18,000) and another, new protein of similar molecular weight. Both the human BR-PO protein (M.W.28,000) and PASII protein (M.W.13,000) showed similar biochemical properties to the corresponding myelin proteins of bovine peripheral nerve, while they both are clearly different from other myelin proteins. Close relationship between the BR-PO protein and the Y protein (M.W.22,000) was suggested by amino acid analysis. Injection of the myelin fraction of bovine peripheral nerve with the complete adjuvant produced EAN while the CNS-BP induced EAE in laboratory animals. However, all three purified proteins, BF-P2, BR-PO and PASII, from bovine peripheral nerve myelin were inactive in inducing demyelinating diseases.


Radioimmunoassay of type D oncovirus from continuous J-96 cells. Radioimmunoassay of J-96 virus and an extract of J-96 cells in the homologous and heterologous systems aimed at detecting antigenic determinants of p25 of Mason-Pfizer virus, as well as group-specific and interspecies antigenic determinants p30 of Rauscher leukaemia virus demonstrated that (1) J-96 virus contains a major internal protein immunologically identical with p25 protein of Mason-Pfizer virus based on the antigenic determinants detectable by the radioimmunoassay used; and (2) no interspecies antigenic determinants characteristic of the major internal protein of mammalian type C viruses were detectable in the J-96 virus or the J-96 cell extract.


The prevention and reversal of digoxin intoxication with specific antibodies. The formation of digoxin-specific antibodies was induced in sheep by immunization with a digoxin-albumin conjugate. The efficacy of the antibodies was investigated in anesthetized cats. When the digoxin-specific antibodies were administered prophylactically as a gammaglobulin, IgG or F (ab')2 preparation, the dose of digoxin needed to induce ventricular dysrhythmia was significantly greater (p less than 0.001) for the pretreated animals than for the controls. To investigate therapeutic efficacy, the animals were digitalized with digoxin over a period of three days and were given digoxin injections on the fourth day to provoke ventricular tachycardia. Of the control animals, three died before two hours had elapsed and the arrhythmia persisted in the two remaining animals. By contrast, a stable sinus rhythm was restored in all animals which were treated with F (ab')2 fragment of the digoxin-specific antibodies after onset of ventricular tachycardia. The doses of digoxin required to trigger renewed ventricular dysrhythmia in these animals were greater than those required at the start of the experiment. The potential clinical use of digoxin-specific antibodies is discussed in the light of these results and reports in the literature.


The cytochemistry of tartrate-resistant acid phosphatase. Technical considerations. Cytochemical demonstration of tartrate-resistant acid phosphatase activity is essential for the diagnosis of leukemic reticuloendotheliosis. In order to perform this test correctly and to interpret the results propertly, it is necessary to understand the technical details of the cytochemical methods thoroughly. The method using naphthol--ASBI phosphoric acid--fast garnet GBC is recommended for this purpose, and factors crucial to the cytochemical study, such as fixation, substrate, coupler, pH and temperature of incubation buffer, counterstains, and mounting media are examined and discussed. Conventional methods for acid phosphatase in the presence and absence of L(+) tartaric acid are also critically examined. The naphthol--ASBI phosphoric acid--fast garnet GBC method is sensitive, technically simple and easily reproducible. Its reaction product is highly chromogenic and is most suitable for cytochemical demonstration of acid phosphatase and tartrate-resistant acid phosphatase activity in cytologic preparations. The naphthol--ASBI phosphoric acid--pararosaniline method is highly specific and is best for histochemical demonstration of acid phosphatase and tartrate-resistant acid phosphatase in tissue sections.


Cytochemical study of estrogen receptor in human mammary cancer. A 17beta-estradiol-6-carboxymethyl-oxime-bovine serum albumin--fluorescein isothiocyanate conjugate is prepared by attaching on the average 11 moles of the fluorescein dye and 24 moles of the steroid hormone to each mole of the protein carrier. This fluorescent estradiol conjugate is used as a tracer to detect estrogen receptor of human mammary cancer cells in frozen sections. The cytochemical findings indicate that mammary carcinomas are composed of heterogeneous populations of receptor-positive and receptor-negative cancer cells in varying proportions and probably should be classified according to the percentages of receptor-positive cells in the cancer cell populations for better correlation with endocrine therapies.


[Postoperative cardiac arrhythmias (author's transl)]. 13 male patients suffering from arteriosclerotic heart disease and/or arterial hypertension were monitored continuously before and after vascular surgical procedures using an arrhythmia computer. Heart rate, paroxysmal supraventricular tachycardias, ventricular extrasystoles, ventricular tachycardias, ventricular fibrillation and prematurity index (QnQe/QTn) were recorded numerically. Ventricular arrhythmias were detected as follows preoperatively in 12 patients, after operation in all patients, paired ventricular extrasystoles or episodes of ventricular tachycardia were found in 5 cases before and in 7 after operation, ventricular fibrillation in one case. The incidence of ventricular dysrhythmias increased significantly (p less than 0.05) early after operation, as did the heart rate during the observed postoperative period (p less than 0.001). The prematurity index dropped below 1.0 during the two days following operation. This differed significantly from the preoperative value (p less than 0.05). The incidence of ventricular extrasystoles was related to postoperative myocardial infarction and heart failure (p less than 0.01), which occurred in 6 cases, with a lethal outcome in three. Only occasionally controlled by trained staff in a normal surgical ward the "Servomed Dysrhythmiemonitor" yielded reliable numerical results during the main part of the monitored period. In two cases it led to immediate detection and rapid institution of treatment of severe tachyar rhythmias.


Gene clusters in the blood group system B of cattle. In this report the linear order of genetic determinants for the blood group system B of cattle has been worked out in more detail. Two groups of antigenic specificities--inclusion groups--showing serologic relationship are described. Recombination frequencies do suggest that these specificities are controlled by two clusters of genetic determinants. The structures of these inclusion groups and clusters are compared with those of antigenic complexes in other species, especially in mice. Several similarities and also dissimilarities between the blood group system B of cattle and the H-2 system in mice are described.


The effect of nasal decongestant on nasal mucous velocity. Nasal mucous velocity was measured in 13 healthy nonsmokers before and 10 min after topical application of 2 alpha adrenergic nasal decongestant sprays, phenylephrine and tetrahydrozoline. Phenylephrine increased nasal mucous velocity from 8.4 +/- 2.7 mm per min (mean +/- SD) to 13.7 +/- 4.8 mm per min and tetrahydrozoline from 8.1 +/- 3.8 mm per min (mean +/- SD) to 13.8 +/- 5.1 mm per min. These increases were significant (P less than 0.01) when compared to topical application of phenylephrine and tetrahydrozoline vehicles, normal saline, and sham (empty aerosol container).


[Food manducation--psychological manducation]. Food "manducation" is the fundamental act of the living process. Through assimilation, the inert nutriment taken from outside the body will wind up as elements making up part of our living being. This merging, incorporating, assimilating process gets unconsciously translated into the everyday language, and provides a share of its vocabulary. It gets likewise translated into various social practices such as greeting and kissing. Those observations in basic anthropology will help substantiating the analysis of pedagogical techniques in the orality civilizations; and particularly--while following in Marcel Jousse's footsteps--in the civilization of that ancient people of the Bible. It comes to light that integrating, memorizing, learning, on the one hand, and teaching, on the other, have but one same psycho-physiological basis. This basis gives rise to a pedagogical system the end and means of which are, on their own plane, a perfect carbon copy of the eating act. Hence, in that ethic group, the analogical parallelism between the physical subsistence, maintained through food manducation, and the intellectual subsistence nourished and enriched through psychological manducation. Thus, such frequently used expressions as: "To eat the lesson", "To eat the teacher while he is instructing", no longer sound obscure. Going beyond, an attempt is made, and this, solely from the anthropological standpoint, to apply these data to the religious and mystical act of Eucharistic Manducation.


An optimised radioimmunoassay for maternal serum alpha1-fetoprotein using polyethylene glycol. A radioimmunoassay for alpha1-fetoprotein in human maternal serum at 15-20 weeks' gestation using polyethylene glycol to separate bound and free antigen is described. A number of variables has been investigated to define optimal conditions for use at this gestation. The method gives results which agree with those from a double-antibody technique. It has good precision in the critical concentration range, and recoveries are satisfactory. The importance of including sufficient serum protein in the analytical system is stressed. There were systematic differences between three standard preparations of alpha1-fetoprotein, but three out of four antisera tested gave very similar results. The implications for screening for neural tube defects in early pregnancy are discussed.


Effect of low concentrations of dihydrostreptomycin on drug resistance in enteric bacteria. Beagle dogs were fed a diet containing 0, 2, or 10 mug of dihydrostreptomycin (DSM) per g of feed. The 2-mug/g level was selected to represent a residue level of the antibiotic. In both treatment groups, medicated feed resulted in a shift from a predominantly streptomycin (SM)-susceptible coliform fecal population to an SM-resistant population. The proportion of resistant organisms was significant (P < 0.01) for both treatment groups. A definitive response did not occur with animals maintained on DSM-free diets. An increase in the prevalence of DSM-resistant organisms was observed after 15 days of DSM-supplemented feeding and persisted during the posttreatment phase of the study. The predominant pattern of resistance was SM-sulfamethoxypyridazine. Fifty-nine percent of SM-resistant strains transferred resistant determinants by conjugation to Escherichia coli K-12 recipients.


Alcohol dehydrogenase from Methylobacterium organophilum. The alcohol dehydrogenase from Methylobacterium organophilum, a facultative methane-oxidizing bacterium, has been purified to homogeneity as indicated by sodium dodecyl sulfate-gel electrophoresis. It has several properties in common with the alcohol dehydrogenases from other methylotrophic bacteria. The active enzyme is a dimeric protein, both subunits having molecular weights of about 62,000. The enzyme exhibits broad substrate specificity for primary alcohols and catalyzes the two-step oxidation of methanol to formate. The apparent Michaelis constants of the enzyme are 2.9 x 10(-5) M for methanol and 8.2 x 10(-5) M for formaldehyde. Activity of the purified enzyme is dependent on phenazine methosulfate. Certain characteristics of this enzyme distinguish it from the other alcohol dehydrogenases of other methylotrophic bacteria. Ammonia is not required for, but stimulates the activity of newly purified enzyme. An absolute dependence on ammonia develops after storage of the purified enzyme. Activity is not inhibited by phosphate. The fluorescence spectrum of the enzyme indicates that it and the cofactor associated with it may be chemically different from the alcohol dehydrogenases from other methylotrophic bacteria. The alcohol dehydrogenases of Hyphomicrobium WC-65, Pseudomonas methanica, Methylosinus trichosporium, and several facultative methylotrophs are serologically related to the enzyme purified in this study. The enzymes of Rhodopseudomonas acidophila and of organisms of the Methylococcus group did not cross-react with the antiserum prepared against the alcohol dehydrogenase of M. organophilum.


[Antigenic relations between human papilloma viruses (HPV): experimental study in guinea-pigs (author's transl]. A study of specific immune response in guinea-pigs after immunization with human papilloma viruses (HPV) purified from plantar warts, hand warts and condyloma acuminata is reported. The respective viruses induced the production of high antibody titres as well as specific cellular immunity as demonstrated by intradermal tests in all the animals. Formalin-inactivated HPV gave similar results. The antigenic cross reactions between the different viruses were evaluated by immunofluorescence and immunodiffusion tests and delayed hypersentitivity reactions. Serologically the results showed viral heterogeneity between condylomata and hand warts, and partial heterogeneity between condylomata plantar warts, and between hand and plantar warts. In skin-hypersensitivity reactions, however, the different viruses appeared as homologous antigens. This experimental study in guinea-pigs showed marked variance of specificity between delayed hypersensitivity reactions and humoral immunity for HPV.


Comparative study of SQ 18 506 with other nitroheterocyclic compounds on experimental Chagas' disease. The activity of the 5-nitrovinylfuran, SQ 18 506, is compared in vivo and in vitro with lampit (a 5-nitrofuran), Ro 7-1051 (a 2-nitroimidazole) and metronidazole (a 5-nitroimidazole). The order of activities on a weight basis was: SQ 18 506 greater than lampit greater than Ro 7-1051 greater than metronidazole. Concern about possible mutagenicity and carcinogenicity, however, has precluded the use of SQ 18 506 in clinical trials.


Oxygen availability, dietary restriction and transport of glucose 3-O-methylglucose and fructose in the isolated small intestine of rat. The influences of PO2 of the incubating medium on glucose, 3-O-methylglucose and fructose transport by everted small intestine sacs in semistarved and rats fed ad libitum (controls) was investigated. Moreover fructose uptake and conversion to glucose by intestinal sacs was also studied. The results showed that intestinal sacs from semistarved rats transported larger amounts of glucose and 3-O-methylglucose and took up more fructose than controls, when PO2 of the incubating medium was 150 mm Hg. There was greater fructose conversion to glucose in the intestine of semistarved rats than in controls at all PO2's considered. The greater functional capacity of intestinal tissue of semistarved rats in comparison to controls has been related to larger O2 availability in their intestinal wall.


Histone inhibition of mitochondrial proton transport. Histone blocks proton uptake by mitochondria incubated in the presence of valinomycin or DNP. In the presence of DNP valinomycin-induced H+ uptake is not affected by histone. H+ uptake induced by nigericin is not affected by histone as well. Postulated mechanism of histone action involves the immobilization of proton translocation in mitochondrial membrane and induction of local change in H+ concentration, the prevention of the interaction between H+ and natural K+-carrier and Mg2+ transport system or valinomycin.


[Complications of cryosurgery in the treatment of prostatic tumors]. Most of the complications in cryosurgery (pyelonephritis, peritonitis, perforations and haemorrhages) are caused by an indequate control of the freezing process or by the use of unsuitable cryosurgical techniques. To this effect, an important role is played by the unsuitable choice of the kind of freezing probe and the use of an unsuitable freezing time. Only endoscopic cryosurgery enables a precise and exact control by means of viewing through the endoscopic trocar along with simultaneous rectal feeling, both of which enable us to achieve an exact supervision of the size of the subvesically-formed freezing sphere adjacent to the rectum. Short, repeated freezings prevent the risk of necrosis (called "sloughts" by English authors). There is a 2% mortality, bearing in mind that the casuistry in cryosurgery is similar to the normal one in open prostate surgery with the difference in favour of the former that these patients generally belong to the so-called inoperable group or with a special surgical risk and aged between 70 and 80 years old. On the other hand, in both groups of our casuistry, the U.T.R. produced a mortality of only 0.3%.


Histone-produced magnesium extrusion from mitochondria and magnesium binding to histone. Histone (60 microgram/mg mit. protein) extrudes Mg2+ from mitochondria by 30% with the utilization of endogenous substrates; in the presence of rotenone extrusion drops to about 18%. Dinitrophenol and ADP prevent this effect of histone. Mg2+ extrusion produced by histone depends on histone concentration being at a maximum (100% extrusion) at 107 microgram histone/mit. protein. It was found also that histone alone binds Mg2+ (1.6 nmol Mg2+/microgram histone).


[Value of alpha-2 globulin, as a biological tracer in the urothelial carcinoma]. We present a series of 63 patients suffering from urothelial carcinoma (53 of the bladder and 10 of the superior urinary tract) in different stages of development. We compare the serum alpha-2 globulin figures with the different parameters making up the TNM classification of the U.I.C.C (period 1974-1979), observing that there is a hyperalpha-2 globulinemia in 40% of the tumours in the superior urinary tract and that, on the other hand, the data obtained for the bladder carcinomas are not conclusive.


Facilitated transport of urea across the gall-bladder luminal membrane. Counterflow experiments demonstrate the existence of urea counter-transport on the epithelium luminal surface. This phenomenon disappears when 10(-4) M phloretin is added to the perfusion fluid. Moreover counterflow experiments made using thiourea as elicitor, demonstrate that the phenomenon is specific for the urea.


Studies on rheomelanins. V. Hemolysis associated with the transformation of catechol into rheomelanin in human blood. Incubation of 2 mg amounts of catechol in 5 ml samples of heparinated blood plasma from four subjects at 38 degrees C for 24 h produced plasma-soluble rheomelanins. These solutions had the brown color and the yellow-green fluorescence in ultraviolet light of 366 nm of other rheomelanins. Their differential ultraviolet and visible spectra showed a rheomelanin absorption maximum at 344 nm. Paper chromatograms of the rheomelanin-plasma solutions in 5% methanol-95% water showed elongated spots of rheomelanins with RF values of 0.82, on Whatman No. 1 paper. Using heparinated distilled water adjusted to pH 7.4 with sodium bicarbonate instead of human blood plasma gave markedly different findings from those obtained with the plasma rheomelanin solutions. Incubation of 4 mg amounts of catechol in 10 ml samples of heparinated whole blood from four subjects for 24, 32 and 48 at 38 degrees C produced rheomelanins as found in the plasma separated from the blood after incubation. The differential ultraviolet and visible spectra of these solutions revealed hemolysis caused by the catechol rheomelanins; this was more marked with longer incubations. The hemolysis was manifested by two absorption peaks at about 270 and 400 nm. Paper chromatography revealed the brown elongated spots of catechol rheomelanins with an RF value of 0.82. Other spots owing to the products of hemolysis were also present.


[Relations between urinary 3-methoxytyramine and the peripheral metabolism of dopamine in human adults and children]. 3-Methoxytyramine (3-MT), the direct metabolite of dopamine (DA) is present in human urines at birth. The rate of its urinary excretion (microgram/24 h) increases highly from birth to 12 months of age; in the next period of infancy, the increase is slower and parallel to that of creatininury; after 15 years, the urinary amounts of 3-MT remain nearly constant, with advancing age. The analysis of the possible sources of urinary 3-MT, lead us to suggest that this compound has essentially a peripheral origin. On the other hand the comparison between urinary data of 3-MT and DA in normal infants and adult subjects has shown that 3-MT excretion is less affected by dietary input than DA itself, so that it can be concluded that urinary 3-MT is a better indication of peripheral dopaminergic activity than urinary DA, chiefly in the young child.


Effects of two different medium-chain triglycerides (tri C8:O and tri C12:O) on liver lipids in the growing rat. Results presented in this study emphasize the long-term effects of dietary fatty acid chain length on some biochemical parameters of the liver in the growing rat. High levels of medium-chain fatty acids feeding (C8:O and C12:O) from 40 to 340 g of body weight induced liver growth and lipid contents intermediary between values recorded with a lipid-free diet and with a diet containing long-chain fatty acids. No steatosis was recorded but neutral lipid contents appeared to be correlated with the dietary fatty acid chain length while phospholipid contents remained much more stable. In the four tested nutritional conditions, only dodecanoïc or lauric acid (C12:O) feeding induced important alterations in the fatty acid pattern of liver neutral lipids. Medium-chain fatty acids, and even lauric acid which was intensely esterified in adipose tissue triglycerides, did not appreciably modify the fatty acid composition of liver phospholipids and were not esterified in it.


Effect of acidosis on heart cAMP-dependent protein kinase. The effect of acidosis on cAMP-dependent protein kinase activity in perfused hearts from normal and reserpinized rats has been investigated. The results were compared to the effect of acidosis on myocardial contractility under the same conditions. The results showed that acidosis increases the cAMP-dependent protein kinase activity in normal hearts. This increase was abolished when the hearts were depleted of norepinephrine by previous treatment with reserpine. As regards myocardial contractility, there was a similar decrease by acidosis either in normal hearts with increased cAMP-dependent protein kinase activity or in reserpinized hearts in which the increase in protein kinase activity was prevented. Two alternative hypotheses are suggested: (1) a dissociation between contractility and cAMP levels, or (2) a "blockade" by acidosis of the mechanical effect of increasing cAMP-dependent protein kinase activity.


[Influence of environmental salinity on the diffusion fluxes of water in the Isopod Sphaeroma serratum (Frabricius)]. 1.--In Sphaeroma serratum, the diffusion fluxes of water are 3 or 4 times more important in sea water (SW) than in diluted (50%) sea water. In 100% and in 50% SW influx equals the outflux. 2.--When the animal is quickly transferred from 100% SW to 50% SW, and from 50% SW to 100% SW, the outflux is instantaneously and entirely reset. 3.--The main factor of the instantaneous resettlement of the outflux seems to be the variation of the concentration of Na+ or Cl- or both together in the external medium; the Ca2+ and Mg2+ concentrations do not seem to have any effect on the diffusion flux. 4.--The validity of our results is discussed (theory of unstirred layers, blood-circulation, oxygen interchange, ultrastructure). Our results are compared with those obtained in other Crustaceans and in Fishes.


Pulmonary vascular changes following air embolism in the dog. The effect of an intravenous injection of air in a dose of 1 ml/kg body weight was determined in 15 healthy mongrel dogs. In 4 control dogs the mean pulmonary artery pressure rose to 2-3 times the resting values at 30 seconds, and carbon monoxide diffusing capacity and pulmonary capillary blood volume decreased by half. In the animals pretreated either with heparin or with methysergide (antiserotonin group) the results were the same as in the control animals. In the vagotomized dogs, the rise in pulmonary artery pressure was not significant, and the decrease in pulmonary capillary blood volume was of lesser magnitude and shorter duration than in the control and the antiserotonin dogs. It is concluded that the intravenous injection of air in supine dogs causes a transient obstruction of small pulmonary arteries. Evidence is presented to implicate a vagal mechanism in both main aspects of the response, namely the pulmonary artery pressure rise, and the partial obstruction of the pulmonary capillary bed. These studies offer additional explanation of the symptoms of respiratory distress observed in rapid decompression.


Kinetics of D-histidine diffusion across rat intestine in vitro. A five-compartment linear model for diffusion in vitro across rat jejunum has been proposed for the study of the kinetic constants of D-histidine transport. Once preliminary experiments using 2,4-dinitrophenol and L-methionine have proved that D-histidine gives rise to passive transport only, the validity of the model was tested and its parameters estimated through a best-fitting procedure by using experimental data concerning D-histidine transport. D-Histidine diffusion was studied in everted and unreverted loops mounted in an oxygenated bath system. Both mucosa to serosa and seroa to mucosa movements of D-histidine (3-30 mM) were evaluated by measuring chemically the amount of D-histidine transported into intestinal lumen every 5 min for 60 min. Results obtained proved that D-histidine transport in each direction (mucosa to seroa or seroa to mucosa) was dependent-concentration process. Nevertheless different values of gain and time constants were estimated for the transport in the two directions.


[Neurotoxicity of acrylamide in rats. I. Subacute intoxication. Absence of effect on incorporation of plasma phosphates into the acid-soluble phosphates of the sciatic nerve and the gastrocnemius muscle, in anatomic continuity and after neural section]. The subcutaneous injection of acrylamide (30 mg kg-1 day-1) in adult male rats induces a severe impairment of the general state of health and a progressive polyneuropathy at the cumulative dose of 180 mg/kg. At the cumulative dose of 400 mg acrylamide does not interfere with the incorporation of plasma inorganic phosphate into the inorganic and organic acid-soluble phosphate fractions of either the gastrocnemius muscle or the sciatic nerve Schwann cells. Nor does it modify the characteristic metabolic response of these fractions to Wallerian degeneration and neurogenic muscle atrophy.


Changes in rodent thyroid hormones and cyclic-AMP following treatment with pineal indolic compounds. Treatment of rats with pineal indolic compounds 5-methoxytryptophol, 5-hydroxytryptophol and serotonin brought about a significant increase in serum thyroxine levels, while serotonin and melatonin caused an increase in thyroid cAMP content with corresponding decrease in the gland's hormones. The total quantity of cAMP in the thyroid was also increased by melatonin in the organ culture system. All these findings would indicate that some of the pineal indoleamines elicit a direct action on the thyroid by stimulating the adenyl cyclase activity and intrathyroidal cAMP, bringing about increased release of thyroxine into the blood stream, and that this is usually not accompanied by adequate synthesis in the gland. Our observation that continuous darkness, which stimulates pineal activity, also brought about an increase in cAMP, concours with our finding of a stimulatory effect of the indolic compounds on thyroid hormone release.


Loss of the lipoprotein lipase activating ability of rat serum after administration of some fatty liver inducing drugs. The effects of the administration of different fatty liver inducing drugs on the serum lipoprotein lipase activating ability was investigated in rats. Addition of serum from 2-mercaptoethanol-, 2-mercaptoacetate-, ethionine- or D-galactosamine- treated rats failed to activate heart and adipose tissue lipoprotein lipase from control rats. The activating effect of serum was only slightly reduced in isopropanol-treated rats, whereas it was found unaffected in ethanol-treated ones. Electrophoresis of the lipoproteins and of the very low density lipoproteins (VLDL) fraction of sera from 2-mercaptoethanol-, 2-mercaptoacetate-, isopropanol-, ethionine- and D-galactosamine-treated rats suggest that the lack of lipoprotein lipase activation ability of these sera is most probably related to the impairing effects of these drugs upon VLDL metabolism, i.e. reduction of VLDL secretion in the case of 2-mercaptoethanol, 2-mercaptoacetate and isopropanol, production of abnormal VLDL in the case of D-galactosamine and both decreased VLDL secretion and production of abnormal VLDL in the case of ethionine.


Unlike effects of denervation on the rate of entry of inorganic phosphate into rat slow and fast muscles. The increased inorganic phosphate flow, characteristic of denervated gastrocnemius muscle is shown to be present in additional denervated fast muscles, i.e. the plantaris, tibialis anterior and extensor digitorum longus muscles. The response of the soleus, a slow muscle, to denervation is biphasic. After an initial decrease of the phosphate flow at the end of the first postoperative day, there is a secondary rise which has the same general characteristics as the rise observed in fast muscles i.e. an exponential or hyperbolic increase to an asymptotic value reached after thirty days. The denervated fast and slow muscles are not converging to an intermediate metabolic pattern. The changes in phosphate flow induced by denervation are reversible in the soleus as well as in the gastrocnemius muscles.


Relationship between the resting blood pressure and depressor responses evoked by stimulation of the aortic nerve. Correlations between the resting blood pressure and magnitude of depressor responses evoked by graded stimulation of the aortic nerve, were studied in rabbits anaesthetized with urethane. The strength of stimulation ranged from 1 to 10 times the threshold, and activated the myelinated afferents. The frequency of stimulation was 5 and 50 cycles/sec. At lower frequency of stimulation the correlations become significant when the intensity of stimulation is 3 times the threshold and they increase at the highest strengths of stimulation. Correlation coefficients calculated for responses obtained at the frequency of 50 cycles/sec are significant in 8 out of 9 intensities of stimulation. With increase in the strength of stimulation they increase, attain their maximum at twice the threshold and decrease at the largest used stimulus strengths. The changes in the values of the correlation coefficients are not paralleled by alterations in the mean size of depressor responses. Since all correlation coefficients are positive, it is inferred that the depressor responses produced by stimulation of the aortic nerve follow WILDER's "law of initial value". The conformity with this rule is the better the higher the values of the correlation coefficients. The strength of relationship between the resting blood pressure and the size of depressor responses is considered to reflect the efficiency of the homeostatic circulatory mechanism.


Influence of salinity on the lipid composition and the fatty-acid pattern of muscle and hepatopancreas of the Chinese crab Eriocheir sinensis. The values of principal lipids and fatty acids from muscle and hepatopancreas of crab E. sinensis living in fresh water or in sea water are determined. Lipid content from these two tissues does not significantly change following a variation in the salinity of the external medium. Nevertheless a slight decrease in the triglyceride concentration (+/- 20%) of hepatopancreas and a very slight increase in the polyunsaturated fatty-acid amount of phospholipids isolated from muscle and hepatopancreas are noted when the crab is transferred from fresh water to sea water.


[Immunological problems of influenza virus infections]. Reference is made to results reported in literature as well as to some of the author's own findings for a summary account of specific immune responses to influenza virus infections. Some introductory comments on the correlations that exist between morphology and function of structural components of the influenza virus as well as on the pathogenesis and its relevance to the immunological responses are followed by a description of the humoral immune response. An assured correlation has been found to exist between the titres of hemagglutination-inhibiting antibodies with responses specific of subtypes and of strains for the protection against homologous influenza virus infection in terms of prevention of outbreak or mitigation of the pathological course. Anti-neuraminidase antibodies have influence upon virus expansion in the organism and in the given population as well as upon the severity of the disease. Particular importance has been attributed to the secretory immunoglobulins. Used in conjunction with interferone, they obviously had an infection-preventing function, wheras the primary role of serum anibodies was a reduction of severity. The studies into cell-mediated immune responses were much less intensive than those conducted into antibody formation. Cell-mediated immune response is thought to emanate from induction of hemagglutinin and is directed against the latter. Sufficient results have not become available as yet on the importance of cellular immune response to protection.


A modified tetrachrome staining technique for the adenohypophysis of dogs, rats and rabbits. A staining technique is described to differentiate between the various types of cell in the anterior pituitary (growth hormone and prolactin producing cells, gonadotrophic, thyrotrophic and ACTY cells) in dogs, rats and rabbits. The method proved to be well reproducible and suitable for the routinely processed pituitary gland. The differentiating characteristics of this method for the prolactin and growth hormone producing cells were confirmed by the immuno-enzyme cytochemical technique in which specific anticanine prolactin and growth hormone sera were used. With this staining technique, the quantitative and qualitative characteristics of the different types of hypophyseal cells at various functional and hormonal stages, under physiological and pathological conditions, can be studied. Effects of progesterone or oestrogen treatment on the morphology of the pituitary gland are illustrated in the photomicrographs.


Antigenic relationship of the feline infectious peritonitis virus to coronaviruses of other species. Utilizing the direct and indirect fluorescent antibody procedure, the antigenic relationship of the feline infectious peritonitis virus (FIPV) to 7 other human and animal coronaviruses was studied. FIPV was found to be closely related to transmissible gastroenteritis virus (TGEV) of swine. Transmissible gastroenteritis virus and FIPV were in turn antigenically related to human coronavirus 229E (HCV-229E) and canine coronavirus (CCV). An interesting finding in the study was that the 8 coronaviruses selected for this study fell into one of two antigenically distinct groups. Viruses in each group were antigenically related to each other to varying degrees, but were antigenically unrelated to coronaviruses of the second group. The first antigenically related group was comprised of mouse hepatitis virus, type 3 (MHV-3), hemeagglutinating encephalomyelitis virus 67N (HEV-67N) of swine, calf diarrhea coronavirus (CDCV), and human coronavirus 0C43 (HCV-OC43). The second antigenically related group was comprised of FIPV, TGEV, HCV-229E and CCV.


Gomori's trichrome stain. Its use with myelin sheaths. We report the use of Gomori's one-step trichrome stain as a method for staining myelin sheaths of peripheral nerves. The stain is simple to perform and allows myelin sheaths to be easily detected and evaluated.


[Citrate and acid phosphatase in the ejaculate in prostatic carcinoma and adenoma]. The simultaneous determination of acid phosphatase and citrate concentration in the seminal fluid obtained from 16 patients with prostatic adenoma showed normal values, 6 patiients with prostatic carcinoma (cytologically and histologically verified) however extremely low values. The difference between persons with prostatic cancer and those with adenoma became particulary obvious with both experimental results evaluated in the way of a twodimensional diagram. This clear separation of both clusters by the simultaneouse estimation of both biochemical parameters may possible get useful diagnostic significance.


[Hyalinosis cutis et mucosae Urbach-Wiethe]. This report deals with the description of a typical case of hyalinosis cutis et mucosae Urbach-Wiethe and with the clinical and histological differential diagnosis to erythropoetic protoporphyria.


Evaluation of toluidine blue for measuring erythrocyte membrane loss during in vivo ageing. A simple method using a cationic dye, toluidine blue (TB), to quantify changes of red cell membrane area has been developed and tested for its validity. After incubating a glucose-depleted red cell suspension with a fixed quantity of TB at 37 degrees C for 10 min, the remaining TB was measured spectrophotometrically at 640 nm. Using this technique, we were able to show differences in TB uptake by populations of young and old red cells. The exact mechanism for TB uptake by the red cells is not clear. Treatment of the cells with bromelin, papain and trypsin reduced the uptake of TB, but neuraminidase and ficin had little or no effect. No inhibition of TB removal by red cells was observed using heparin, D-glucose, glucuronic acid, or N-acetylglucosamine.


Transcription of the alpha2u-globulin gene in male rat liver nuclei in vitro. Alpha2u-globulin is a male rat liver protein under multihormonal control which represents approximately 1% of hepatic protein synthesis. We have measured the rate of transcription of the alpha2u-globulin gene using a nuclear cell-free transcriptional system with mercurated CTP as substrate for the endogenous RNA polymerases. The newly synthesized, mercurated RNA was purified free of endogenous RNA by chromatography on sulfhydryl agarose and hybridized to 3H-labeled alpha2u-globulin cDNA. It was found that, in male rat liver nuclei, alpha2u-globulin RNA sequences represent 0.005% of the total newly synthesized RNA. Actinomycin D or alpha-amanitin completely blocks the synthesis of alpha2u-globulin RNA in these nuclei. No alpha2u-globulin RNA synthesis was detectable in female rat liver nuclei. Thus, within the limits of our detection the absence of hepatic alpha2u-globulin mRNA in female rats appears to be due to a lack of transcription of the alpha2u-globulin gene in these animals.


A critical examination of possible fractionations of human DNA according to base composition. Human DNA has been fractionated according to base composition by sedimentation equilibrium in an HgCl2/Cs2SO4 density gradient, followed by sedimentation equilibrium in an actinomycin/cesium formate density gradient. The fractions of different base composition resulting from this procedure were subsequently analyzed by sedimentation equilibrium in CsCl, DNA renaturation kinetics, and electron microscopy. All fractions contain similar kinetic classes of repeated DNA sequences as judged by renaturation studies. Short (300 nucleotides) interspersed repeated sequences are found in all fractions with no noticeable enrichment for these sequences in any fraction. Repeated sequences from fractions of different base composition are partially able to cross-hybridize, demonstrating that nearly identical repeated sequences occur in molecules of different base composition. These findings are critically compared to reports of successful density gradient fractionations of different human DNA sequence classes.


Reductive dechlorination and degradation of mirex and kepone with Vitamin B12. Vitamin B12s effects the reductive dechlorination of mirex (dechlorane) in protic solvent systems, under both catalytic and stoichiometric conditions, mainly to yield compounds of composition C10Cl12-nHn, with n = 1-8, in which the basic dihomocubane cage structure is retained; the formation of cage-opened, reductively dehalogenated derivatives of 4,7-methanoindene occurs only to a very minor extent. The corresponding reactions of kepone (chlordecone), in contrast, occur with predominant formation of indene derivatives. Under certain mild conditions, vitamin B12s induces a fragmentation of kepone leading to the destruction of the dihomocubane moiety and the formation of an isolable organocobalamin having a C3Cl3H2 residue attached to the cobalt atom. In strongly alkaline media, the reaction of kepone with vitamin B12s may in addition yield high-molecular-weight condensation products of unknown constitution. Reactions of this type are of interest as prototypes of soil-decontamination processes.


Serum alphafetoprotein in multiple pregnancy. The concentration of alpha-fetoprotein (AFP) was determined in paired umbilical cord and maternal sera in 42 multiple pregnancies. No concentrations above 1.4 microgram/ml were detected in maternal sera. Although there was a significant inverse correlation between cord AFP levels and gestational age, large intrapair discrepancies were common and these were not influenced by birth order, weight, or malformations. Intrapair AFP ratios were higher amongst dizygotic (DZ) than monozygotic (MZ) twins. In a pair discordant for neonatal hepatitis, the affected twin had the lower level of AFP in cord serum, but AFP was still detectable at 55 days.


[Effect of acetylcholine on the pituitrin induced osmotic flow of water through the wall of the frog urinary bladder]. The role of intercellular pathways in the ADH-dependent water transport was studied on the frog urinary bladder by means of acetylcholine (AC) and other cholinergic compounds. AC (10(-3) M) was found to cause a strong suppression of the pituitrin-stimulated water flow. Analogous effect was produced by AC on the osmotic flow stimulated by cyclic adenosine monophosphate (cAMP) and theolin. The antipituitrin effect was not reproduced either by nicotine, nor by potent M-cholinomimetic agents (methylfurmetide and F-2268), and was not prevented by M- and N-cholynolytic drugs (atropine, metacin, flaxedil, hexamethonium). However, the antipituitrin effect of AC was completely removed by the anticholinesterase drugs with different mode of action (eserine, proserine, armin, acridine iodmethylate, GD-42) in concentrations of 10(-6)--10(-3) M. It was concluded that the smooth muscles contraction with the subsequent closure of the intercellular spaces was not responsible for the antipituitrinic action of AC. This effect appears to be connected with cholinesterase activation. A possible role of the phosphoinositides in the water permeability regulation of the urinary bladder wall is discussed.


[Size of the "distortion" area of the membrane close to the ion channel formed by gramicidin A]. Young's modolus of the bilayer membrane (BLM) in the direction normal to membrane plane approximately E perpendicular changes if gramicidin A molecules are incorporated into BLM. The plot of approximately E perpendicular on membrane conductivity is a monotonic curve with saturation at BLM conductivity corresponding distances between gramicidin A dimers about 1000 angstrom. The conclusion is made that dimer formation generates drastic reorganization of BLM structure, the size of distortion zone of the membrane near the ionic channel is of the same value.


[Antigenic marker of human erythrokaryocytes similar to mouse erythroblastosis antigen]. Identical antigenic determinants are discovered on the surface of human erythrokaryocytes with antibodies against specific antigen of murine erythroblasts (Ag-Ed), previously revealed in study of Rauscher leukemia, in the immunofluorescent and cytotoxic tests. The antigen is present on the membranes of the majority of human embryonic liver and adult bone marrow nuclear erythroid cells, but is not found in fetal thymocytes, newborn kidney cells, adult human hepatic cells and in peripheral blood erythrocytes. Ag-Eb appears to possess an inter-species determinant, shared by mammalian nuclear erythroid cells, and may be used as their specific marker.


[Comparison of two means of identifying satellite associations in man]. Two methods of identifying satellite associations were compared in ten normal donors. Conventional Giemsa stain proved to identify by 31% less of 3-chromosomal associations, by 50% less of 4- and 5-, and by 66.6% less of 6-chromosomal associations than in staining with ammoniacal silver, whereas the number of 2-chromosomal associations was by 10% greater, this conditioning the prevalence of small associations. Silver staining used to expose intersatellite associations revealed acrocentric chromosomes mutually oriented, placed close to one another, but without any intersatellite connection. In Giemsa-stained preparations these are considered to be associating, which increases the number of multiple groups. Both errors of the conventional Giemsa method cannot be corrected. Identification of associations according to intersatellite connections is free of these errors after silver staining.