Relationship between alpha-L-fucosidase deficiency in plasma and alpha-L-fucosidase activity in leukocytes. After testing a population sample of 185 hospitalized Italian children for the plasma alpha-L-fucosidase deficiency and establishing an approximate threshold value between heterozygotes and wild-type homozygotes, we analyzed by two statistical methods the distribution of the two genotypes. The results obtained by probit analysis agree with threshold and average values expected on the basis of the Hardy-Weinberg equilibrium. In addition, the level of alpha-fucosidase in leukocytes of 12 individuals with deficiency of alpha-fucosidase in plasma was found to be significantly lower than that of 61 controls (P less than 0.005). These results indicate that the mutation(s) causing a deficiency of alpha-fucosidase in plasma is (are) also expressed in leukocytes.


Characteristics of galactokinase and galactose-1-phosphate uridyltransferase in cultivated fibroblasts and amniotic fluid cells. The kinetic characteristics of galactose-1-phosphate uridyltransferase and galactokinase in cultivated fibroblasts and amniotic fluid cells were investigated. The Km values of galactokinase for galactose at 2.0 mM ATP are 0.34 mM in amniotic fluid cells and 0.48 mM in fibroblasts. The Km values for ATP at 0.5 mM galactose are 1.25 mM and 2.10 mM. Transferase and galactokinase activities and protein content increase logarithmically during the growth of cultivated cells. The specific activity of both enzymes also increases and reaches a maximum level 10--15 days after subculture. The specific activity of transferase increases faster than that of galactokinase in the case of amniotic fluid cells. In the case of fibroblasts the specific activity of galactokinase increases faster than that of transferase.


The constitutional fragility of chromosome 12 in a case of 46,XX,var(12)(gh',RHG,CAG,CBG). The constitutional fragility of chromosome no. 12 in a female infant with unspecific clinical signs is described. RHG, GAG, and CBG methods were used to localize the fragile point. The breaks seem to be in 12q1.3, and always within an R band. A possible correlation between the phenotypic modifications and the chromosome variant is discussed.


18Q - syndrome resulting from a tdic(14p; 18q). A case of 18q- syndrome due to a de novo tdic(14p;18q) is presented. The interest of this observation lies in the rarity of stable dicentric chromosomes arising from reciprocal translocations between autosomes.


Abnormal childhood phenotypes associated with the same balanced chromosome rearrangements as in the parents. Three patients have been detected because of abnormal phenotypes. Balanced chromosome rearrangements were found in their karyotypes, and in one of the parents in each case. In these three families the association of an abnormal phenotype and a balanced chromosome rearrangement may either be a chance happening or be induced by a submicroscopic loss of chromosome material, for which different position effects may be responsible, according to the case in question. These observations prompt us to ask what decision should be made relative to termination of pregnancy if such a rearrangement is found in amniotic cells in a family with a previous abnormal child.


Sister chromatid exchange and cell cycle in fibroblasts of Bloom's syndrome. Sister chromatid exchange (SCE) has been studied in the fibroblasts of five Bloom's syndrome patients, one heterozygote, and two normal individuals. The high frequency of SCE already known in the lymphocytes of Bloom's syndrome was also found in the fibroblasts of all five patients. However, populations with low and high frequency of SCE were not found. In addition, chromosome aberrations appeared with a lower frequency. The cell cycle duration in the Bloom's fibroblasts appeared to be similar to that in the normal cell line, and the difference in the growth pattern appeared to be due to the variation in the mitotic index. The cell cycle lasted about 24 h in at least four of the Bloom's lines studied during the present experiments.


Single Cd band in dicentric translocations with one suppressed centromere. Six human dicentric translocations involving the presence of a suppressed centromere were studied: one case of t dic(9;22), one of t dic(13;18), one of t dic(14;15), and three of t dic(13;14). All exhibited chromatid separation at an acrocentric centromere and were demonstrated to have two regions of centromeric constitutive heterochromatin. The four that were available for retrospective study showed a single Cd band, with absence of the Cd band at the suppressed centromere.


Terminal long-arm deletion of chromosome 1 in a male infant. The clinical findings in a child with a terminal deletion (1) (q42 leads to qter) is described and compared with three cases with a similar chromosomal anomaly.


Three cases of ring chromosome 2, one derived from a paternal 2/6 translocation. Two cases of primary ring chromosome 2 and one case of a ring secondary to a paternal 2/6 translocation are described and compared with a fourth case of ring 2 from the literature. The breakpoints in two cases are identical and the same as the breakpoint on chromosome 2 in the composite 2/6 ring. The three primary rings have a number of symptoms in common, but other cases are needed to identify a possible deletion syndrome. The levels of the enzyme MDH-1 were within the normal range in Cases 1 and 2. Thus we localised the locus for MDH-1 to segment 2p23 leads to p25; this had previously been localised to segment p23 leads to pter.


Sex-linked inheritance of cleft palate.

Sex-linked inheritance of cleft palate. A family pedigree is reported in which males in four generations are affected with midline isolated cleft palate. This is the second report of cleft palate inherited as a sex-linked recessive trait, and it emphasizes the importance of careful pedigree construction when counseling families of children afflicted with this disorder.


Translocation of a supernumerary Y to a 15: study of six cases (three males and three females) in three generations. We present a family in which six members (three males and three females) carry a translocation of the fluorescent part of a supernumerary Y to a 15. The carriers do not have any impairment of fertility or any malformation.


Investigation of associations between ABO blood groups and coagulation, fibrinolysis, total lipids, cholesterol, and triglycerides. To investigate possible associations between ABO blood system and coagulability levels, fibrinolysis, total lipids, cholesterol, and triglycerides, the plasma and serum of 300 Rh-positive male blood donors were tested. The tests performed were: RT, PTT, K-PTT, PT, F.V, F.II, F.VII, Complex II, VII, and X, TGT, fibrinogen, HAE 0.2, HAE 0.5, ELT, LIP, Col.1, Col.2 and TRI. Analysis of the laboratory data shows a lower coagulability in O blood group individuals. This result was obtained in coagulation tests (RT, PTT, and K-PTT) specific for factor VIII level. In addition, a higher sensitivity to the in vitro heparin anticoagulant effect in O group individuals was confirmed. Nevertheless, these conclusions are specific for Negroids, the same effects not being observed in Caucasians. None of the other laboratory tests revealed any differences related to either blood group or race.


The occurrence of DChi and new D and B transferrin variants among caste groups of Andhra Pradesh (S. India). Serum samples from a total of 862 unrelated individuals belonging to seven Hindu endogamous caste groups residing in Hyderabad and Warangal cities of Andhra Pradesh were examined for electrophoretic variation of transferrin (Tf). In four caste groups, namely, Brahmin, Vysya, Padmashali, and Kapu, the Mongoloid genetic marker DChi was found, with polymorphic frequencies in Brahmin and Vysya groups. The two new D variants found in individuals of Madiga and Mudiraj castes were designated as DMadiga and DMudiraj. Similarly, two new B variants found in individuals of Goldsmith and Madiga castes were designated as BGoldsmith and BMadiga, respectively. These findings suggest a widespread transferrin variation in caste groups of Andhra Pradesh and in the tribal populations of this region.


Direct duplication 2p14 to 2p23.

Direct duplication 2p14 to 2p23. A malformed male newborn was first diagnosed as having Smith-Lemli-Opitz syndrome. Extensive cytogenetic studies, including Q, G, C, R and T banding and BudR treatment, were applied, finally leading the authors to conclude that the patient had a partial 2p trisomy caused by direct duplication 2p14 to 2p23. This was a de novo chromosome abnormality, as both parents had normal karyotypes.


Partial trisomy 13 as a result of de novo (6p;13q) translocation. A newborn infant with the clinical features of the Patau syndrome was found to have excess chromosome 13 material present as a tandem translocation involving the short arm of chromosome 6 and the long arm of an extra chromosome 13: 46,XY,t(6;13)(p24;q12). The major part of the long arm of the extra chromosome 13 was attached linearly (tandem translocation) to the short arm of chromosome 6. Both parents were phenotypically and karyotypically normal.


Decision on the control of a dietary carcinogen -- aflatoxin. The sequence of events is described whereby decisions are made by the scientific community and national or international legislative bodies in evaluating a dietary carcinogen. This is proposed on the basis of experience gained from the aflatoxins. Suspicion arose first after the deaths of large numbers of poultry and fish; the agent was soon identified as aflatoxins. This was followed by laboratory investigations of their toxic and carcinogenic effects in experimental animals, which showed them to be the most potent liver carcinogen known to experimentalists. Field studies were established to assess a possible association with liver-cell cancer in humans, and a significant correlation was found. An intervention programme was therefore set up to lower exposure and to observe the trend of cancer incidence. Aflatoxins are found in many cereals and nuts exposed to humidity; thus, the staple foods of most of the developing world are at risk. The severity of the economic problem for some countries if prejudice should continue to develop against peanuts as a basis for animal feeds is illustrated. Decisions about control of aflatoxins thus fall into two groups: control measures to prevent contaminated foods from reaching the populations of industrialized countries, and primary prevention for the millions in the developing world. Regulatory actions have been taken in various industrialized countries, but their effect is to make developing countries safeguard exports and still to ignore the hazard within the country. The main problem is thus protection of food supplies, for the indigenous populations of those countries that grow and consume the bulk of the susceptible cereals, by improved husbandry.


Residues of carcinogenic animal drugs in food: difficulties in evaluation of human safety. The indisputable need to intensify animal production in order to provide an adequate food supply for the world population involves the use of substances that are highly potent pharmacologically and toxicologically. The history of regulatory action with regard to such additives is similar to that for other substances: first, no regulation; next, an over-reaction; and now decisions based on judicious evaluation of scientific facts. One factor that differentiates the chemicals used in animal production from other food additives is that both the parent compounds and their metabolites appear in edible products, posing problems both for the analytical detection and safety evaluation of such residues. It would be unrealistic to propose 'zero' tolerances for these additives, even if they are carcinogenic. The benefits gained from drugs that cure and prevent infections and parasitic diseases in food-producing animals, and the fact that analytical methods can now detect very small quantities make the presence of low levels of these substances in food unobjectionable.


Carcinogenic risk of products used in the pharmaceutical and related industries. The pharmaceutical industry is perhaps most subject to rules and regulations in the majority of countries. Legislation in France in this regard is flexible and relatively easily adaptable; in addition, the pharmaceutical industry has established its own ground rules. One of these requires that a product be harmless under normal conditions of use; carcinogenicity checks are only mandatory under certain conditions. If they were to be made compulsory, a less unwieldy test system would be required. The use to which a product is put should be taken into consideration in regulating it.


Review of pesticide carcinogenesis data and regulatory approaches. Carcinogenicity data are presented for compounds used as pesticides which have been evaluated in the IARC Monograph series. These pesticides have been both poorly and scantily tested for carcinogenicity. Additional tests have been carried out under the NCI Carcinogenesis Program, and detailed results of the bioassays are published in the NCI Carcinogenesis Technical Report Series. A prpcedure was developed whereby 'preliminary reports' could be made in the Federal Register. In the second part of the paper, regulatory approaches to pesticides are described. International activities are outlined; and examples are given of factors, such as public concern, scientific work and governmental response, which are highlights in the development of approaches to such regulations. The problem of risk assessment has been approached from many sides but remains largely unsolved. Besides risks and benefits, an essential component in evaluation is the analysis of technological alternatives. Two other factors are public documentation of the process of evaluation and registration of environmental carcinogens and their uses. Reduction of exposure to environmental carcinogens to a 'minimum feasible level' depends on social, economic and temporal factors and implies the acceptance of a finite level of risk.


Research related to the herbicide buvinol, especially for possible carcinogenicity. The Budapest Chemical Works (Hungary) has conducted a study on a new herbicide called 'Buvinol', containing 2,4,5-trichlorophenoxyethanol (TCPE). In long-term carcinogenicity testing, the oral administration of the maximum tolerated dose of TCPE (70 mg/kg), containing 0.1 ppm 2,3,7,8-tetrachlorodibenzo-para-dioxin, increased the incidence of liver tumours in male mice, while smaller doses (1/10 and 1/100) did not. Dioxin in the dose range used (0.1--10 ppm) did not influence tumour frequency. Exposure of workers to TCPE during its production or use should be controlled and reduced.


The role of laboratory animal studies in estimating carcinogenic risks for man. The extent to which biological processes predict those in humans is discussed and illustrated by analysis of data presented in the first 16 volumes of the IARC Monograph series. Other examples are given to show that if there is sufficient evidence that a chemical is carcinogenic in appropriate animal test systems it must be treated as if it were carcinogenic in humans. A quantitative correlation between data in animals and in humans is more difficult to establish, although there is tentative evidence that such a relationship exists. Society should attempt to keep 'inevitable' exposures to carcinogens to a minimum; social need should be balanced against social risk. Biomedical research can help to estimate this role.


An overview of the problem of thresholds for chemical carcinogens. Toxicological evaluation in quantitative terms of substances that have been shown to be carcinogenic is very difficult, mainly because, for the great majority of environmental carcinogenic chemicals, the available data, both from experimental studies in animals and from epidemiological studies in humans, on dose-effect relationships, especially at low levels, are very limited. There is an urgent need for research in this area. At present, the most sensible approach is to try to determine no-effect levels by long-term investigation of the effects of low levels in animal species that are most similar to man in their metabolic potential and their sensitivity. Very low safety factors (2500 to 5000) should be applied to the values thus obtained, in order to be as far as possible on the safe side in establishing pragmatically acceptable limits. At present, since all chemical carcinogens pose a hazard, human exposure to them must be reduced to the feasible minimum.


'Human' and 'animal' carcinogens.

'Human' and 'animal' carcinogens. A clarification is made of the meanings of the terms 'human carcinogen' and 'animal carcinogen' in cancer research, cancer prevention and legislative actions. A chemical proven to have carcinogenic activity in animals should be considered to represent a risk to humans. However, non-experts tend to consider that 'animal carcinogens' produce cancer only in animals and can be neglected with regard to the human situation. The use of these terms should thus be avoided or they should be used as standard terms based on generally accepted and available definitions.


Anti-smoking legislation in Finland.

Anti-smoking legislation in Finland. The act which outlined measures for restricting tobacco smoking in Finland took effect on 1 March 1977. This act is presented and its background described. The main regulations provided by the law are: 1) Health authorities administer all tobacco regulation, except taxation; 2) 5% of tobacco taxes are alloted annually to health education research and evaluation; 3) advertising and promotion of tobacco are prohibited; 4) limits on tar, nicotine and carbon monoxide are set by the government; these must be met for a permit to be granted; 5) tobacco products are quality controlled; 6) cigarette packets must bear health warnings; 7) no tobacco products may be sold to persons under sixteen years; 8) they may be sold only from supervised dispensing machines; 9) smoking is prohibited in all public areas, including transport; 10) the Board of Health administers nation-wide education measures. With regard to the latter regulation, some past and projected programmes are described. Initial results of the campaign were limited; further support from taxation and a pricing policy is necessary.


alpha, beta Mannitone, a new industrial material with unknown benefits and risks. The example is given of a new, incompletely characterized material about which information is currently incomplete. alpha, beta-Mannitone is made from old automobile tires by a secret process; it is not known whether worker exposure will occur during its production; and it is proposed for use as a substitute for petrol. The risks and benefits of this hypothetical substance must be weighed. The problems associated with such an evaluation are listed in the form of questions.


Legal forms of intervention against carcinogenic risks. The place of jurists in the formulation of regulations against carcinogenic risks is at the end of a long chain forged by interdisciplinary cooperation. Legal processes for intervention can be brought about in a number of ways: 1) Civil actions; 3) compensation; 3) preventive regulations; 4) regulations specific to the product, its production or its use; 5) institutions to provide surveillance of application of the regulation, for emergency interventions, sanctions against offenders, appeal bodies and flexibility; 6) international cooperation; 7) international trade agreements; 8) formulation of international regulations; 9) use of existing international regulations; 10) development of international rules for testing new products; 11) reference to existing regulation.


The Clearinghouse for Environmental Carcinogens of the National Cancer Institute, USA. The NCI Clearinghouse for Environmental Carcinogens acts as an advisory body to the Carcinogenesis Testing Program of that Institute. It consists of 30 members, from academic institutions, labour organizations, consumer groups and industry. They are divided into three groups: the Chemical Selection group establishes priorities for compounds to be tested; the Experimental Design group decides on the test procedures and parameters to be used; and the Data Evaluation/Risk Assessment group review data from completed tests, for deficiencies in experimental design or conduct, the nature of the lesions produced and the statistical analysis of the results, and gives a critical opinion of the test.


Techniques in intervention against carcinogenic hazards. The principal techniques for the control of carcinogenic hazards are described. A sense of responsibility must be encouraged in the groups of people who are involved, such as manufacturers, importers, retailers or distributers. Relevant, evaluated information from the scientific community must be brought to the attention of responsible bodies in a systematic way. Information about sources of emission, pathways of spread and identification of populations at risk is essential. Estimates must be made of costs and benefits, whether tangible or intangible. Mechanisms of intervention are described, including registration, authorization, permission, notification and dissemination of information. Mechanisms of risk surveillance involve the monitoring of health indices and of exposure situations: techniques of data collection are outlined. It is important that such data be both accessible for evaluation by authorized persons and at the same time remain confidential. Education is basic to cancer prevention. National and international coordination is essential. The paper concludes with a list of areas in which strategies and techniques should be developed.


Implementation of regulations and promotion of standards for protective measures in the absence of statutory regulations. Asbestos regulations in the UK and several other countries are described, and it is suggested that these could be used as a basis for similar regulations in other countries which do not yet have them. Those aspects of the UK regulations which make them particularly applicable to all occupational situations in which asbestos dust might be emitted are listed. As a result of such regulations, standards of control have improved; two examples are given.


Assessment of trichloroethylene as an occupational carcinogen. The reaction is described of the National Institute for Occupational Safety and Health (NIOSH) in the US to the results of experimental studies which showed that tetrachlorethylene is carcinogenic. Parallels are given with vinyl chloride, which it resembles structurally. The main occupational exposure to trichloroethylene is in the vapour degreasing of metal parts. The machine used for this activity is described, and it is shown that there is a high potential exposure. Possible control measures are outlined. The main evidence for the potential carcinogenicity of trichloroethylene is the occurrence of its epoxide as an intermediate in its metabolism to chloroethanol; species differences have been observed. Epdemiological studies have so far been inconclusive. NIOSH decided that trichloroethylene is potentially carcinogenic in the workplace, but only weakly so. No new standard has been recommended, although a lowering of the present standard is recommended.


Implications of health and safety legislation. The UK Health and Safety at Work Act is described; the duties of the employer, including manufacturers, importers and suppliers, and of the employee for health and safety are laid down. This Act provides the necessary legislative framework for the control of carcinogenic, mutagenic and other health hazards at work. Proposed regulations dealing specifically with carcinogenic substances and with the notification of toxic properties will add strength to these general provisions. The implementation of the new regulations will be the responsibility of the Health and Safety Commission, advised by an expert subcommittee. Ad hoc decisions about handling known and suspected carcinogens are made in consultation with employers and employees. For practical purposes, there must be working limits for carcinogens; zero levels and 'lowest levels of detection' are impracticable. The difficulties of defining the criteria by which substances are judged to be carcinogenic to humans are outlined. Science plays only one part in decision-making, and final decisions depend also on social and economic needs.


Tobacco--current French legislation.

Tobacco--current French legislation. A recent French campaign to bring about a reduction in tobacco consumption is described. A number of factors were used to convince the public that smoking is dangerous: it was pointed out that, of all cancers, those in which tobacco plays an important role are increasing the most rapidly; another is that smoking reduces life expectancy.  A study of the smoking habit in France showed that it is acquired earlier and earlier; another study showed that 60,000 deaths a year can be linked to smoking; the cost of medical services related to smoking was calculated to be 22 thousand million francs. By comparison, the budget of the anti-smoking campaign was 3 million francs, but this was sufficient to reduce the risk by 7%. The second objective of the programme was to give non-smokers the possibility to assert their rights. The main target of the campaign was young people. The social pressure on them to begin smoking could be reduced if the number of smokers were reduced, if non-smokers asserted their rights and if the image of the smoker were debunked. The campaign was supported by anti-smoking legislation which came into effect in July 1976. Smaller campaigns were planned to follow annually. The main targets of the law were advertising and publicity; in addition, smoking was prohibited in public places, public means of transport and educational establishments. The effectiveness of the campaign was measured by sampling changes in behaviour and in sales volume: there was a 5% decrease in the number of smokers and a 15% decrease in consumption of those who continued to smoke.


Regulation of carcinogenic food additives and drugs in the US. The four components of the process by which a decision is made about the acceptability of human exposure to a chemical are outlined. The quantity and quality of the data that are required have increased considerably in recent years, and the standards governing the conduct of the studies done to obtain them have become more stringent. Thus, many chemicals now in widespread use were admitted on the basis of evidence that is now considered woefully inadequate; the steps being taken in the US to remedy this situation are described. Since such testing is long and expensive, four regulatory agencies have joined together to develop guidelines that can be used by each. The interpretation of data presents yet another problem. Further, what constitutes adequate evidence for safety is a matter subject to wide ranges of opinion: this is illustrated by the example of cyclamate. The difficulties of extrapolating results to actual situations are outlined; and the limitations of the usefulness of experimental testing are pointed out. Determination of an acceptable ratio between benefits and risks depends on a wide variety of factors, and regulatory decisions in this regard thus depend on several kinds of scientific judgements and on policy decisions based on cultural aspects.


Effects of some antibiotics on the growth of human diploid skin fibroblasts in cell culture. During serial subcultures 50 micrograms per ml gentamicin and penicillin (100 U per ml)-streptomycin (100 micrograms per ml) depressed cell growth signficantly 2 weeks after the addition of the antibiotics; gentamicin, but not penicillin-streptomycin, stimulated cell growth before it became inhibitory. Removal of the antibiotics resulted in the cell yield returning to normal. The results show that these antibiotics can be harmful to cells even at concentrations thought to be safe.


Differentiation in human amniotic fluid cell cultures: chorionic gonadotropin production. Two of the distinguishable cell classes subcultured from human amniotic fluid were examined for their capability to produce human chorionic gonadotropin (hCG) as determined by radioimmunoassay. The class that predominates in most cultures used for prenatal genetic diagnosis, previously termed AF (for amniotic fluid), secretes hCG into the culture medium. Dermal fibroblasts do not, nor does another type of cultured cell from amniotic fluid, previously termed F because of a resemblance to fibroblasts. Primary AF cultures produce more hCG than do subcultures. Evidence that this hormone is intact hCG is provided by its immunoreactivity with antisera raised against the beta-subunit and against the intact molecule of hCG. Furthermore, a dose-response curve for hormone in culture medium is parallel to that of highly purified intact hCG. It is postulated that AF cultures are derived from fetal membranes and retain properties of trophoblast.


Factors influencing microbiological assay of cell-culture mycoplasma. A total of 6432 cell cultures was assayed for mycoplasmas over a 6-year period by aerobic and anaerobic incubation of agar and broth media. Mycoplasmas were detected in 375 cultures (5.8%). M. orale and A. laidlawii accounted for 61.3% of the isolates. Anaerobic incubation detected 98.1% of the isolates; aeorbic incubation detected 45.8%. Of factors studied to determine their effect on mycoplasma assay, only two, anaerobic incubation and presence of mycoplasmacidal/static antibiotics, were significant. In separate studies, 86 of 2656 cell cultures (3.2%) were infected with strains of M. hyorhinis that did not grow on cell-free media. Recommendations are given for microbiological assay of cell-culture mycoplasmas.


Rimantadine but not amantadine protects Fischer rat embryo cells from transformation induced by 3-methylcholanthrene or benzo(a)pyrene. The antiviral drugs amantadine hydrochloride and rimantadine hydrochloride were tested as to their oncogenic potential using a serial line of Fischer rat embryo cells that previously had been shown to be an accurate indicator of chemicals known to be oncogenic in animal studies. Neither compound was found to have transforming activity. At slightly toxic levels, rimanbadine hydrochloride, but not amantadine hydrochloride, protected the same cell line from the transformation induced by the polycyclic hydrocarbons 3-methylcholanthrene and benzo(a)pyrene.


The development of mouse spinal cord in tissue culture. I. Cultures of whole mouse embryos and spinal-cord primordia. Whole mouse embryos were grown in vitro from Theiler stage 12 (1 to 7 somites) to Theiler stages 15 and 16 (25 to 35 somites). This procedure gives experimental access to precisely staged embryos during the early period of neurogenesis. To follow the further development of neurons in vitro, fragments of spinal primordia were set up from these cultured embryos. In such cultures, the proliferation of precursor cells, the formation of postmitotic cells, and, finally, the cytodifferentiation of neurons were observed.


In vitro migration of epidermal cells in suction blisters of rat skin. Wounds of the external ear of the rat created by suction were carried in vitro up to 48 hr, and the growth of epidermal cells was observed by scanning and transmission electron microscopy. Epidermal cells migrated on the intact basal lamina taking origin from the surrounding uninjured epidermis and from the external root sheaths of hair follicles. The time required to form a confluent layer of cells was much shorter than that observed earlier in intact blisters under in vivo conditions. This model offers promise for the further study of the migration of epithelial cells.


Organ culture of adult mouse intestine. II. Mitotic activity, DNA synthesis and cellular migration after 24 and 48 hours of culture. DNA synthesis, mitotic activity and cell migration have been measured in organ culture of adult mouse jejunum during the first 48 hr. Explants cultured in DMEM-HEPES-NCTC-135 medium present a sharp decrease of mitotic activity in the first hours of culture, but mitoses are restored to 80% of the controls between 24 and 48 hr. DNA synthesis follows the same pattern. Cell migration continues during culture.


Organ culture of adult mouse intestine. III. Behavior of the proteins, DNA content and brush border membrane enzymatic activities. The proteins, the DNA content and the brush border membrane enzyme activities of adult mouse intestinal explants have been measured during a 24-hr organ culture. These activities were not modified in comparison with the controls at the beginning of the culture. The activity of the enzymes secreted in the medium during the 24-hr culture was equal to the activity present in the explants at the start of the culture. These results show that several metabolic functions of the intestine are fully preserved in organ culture.


[The mycoplasma-pneumonia. A model for the study of the pathogenesis of infectious diseases (author's transl)]. The known mechanisms of pathogenicity of Mycoplasma pneumoniae are discussed. In addition, evidence is provided for the participation of the immune system in the pathogenesis of M. pneumoniae disease. In a working hypothesis the stimulation of a protective and/or a reactive immune response is postulated, depending on the predisposition of the host, the infecting dose, the frequency of contact and the virulence of the organisms. Because well studied animal models are available, the Mycoplasma-Pneumonia is well suited to investigate the pathogenesis of infections caused by bacteria and the defense of the host.


[Phagocytic-activity in high risk patients of a surgical intensive care unit (author's transl)]. The in vitro phagocytic activity from PMNL against eight different Klebsiella strains, isolated from clinical material, was investigated and the K-index calculated for fifteen patients from a surgical intensive care unit. The hemagglutinating antibody titers against the test strains, as well as the serum concentrations of IgM, IgG, C3 and C4 were also determined. At the time of the investigation all patients had symptoms of a severe general infection. There could be no decrease in the phagocytic activity demonstrated. However, large variations against certain bacterial strains were observed. This same strain dependency was also observed for five healthy persons. The hemagglutinating antibody titers of any one patient were either generally increased for all strains or uniformly decreased. The serum concentrations of IgM, IgG, C3 and C4 were found to be, in general, within the normal large.


[Cefoxitin and cefuroxim concentrations after intravenous administration in sputum of patients with chronic bronchitis (author's transl)]. On 24 patients with chronic obstructive bronchitis the Cefuroxim and Cefoxitin concentrations in the sputum were investigated during 11 days after intravenous injections. The concentrations of Cefuroxim were above the minimal concentrations necessary to block growing of bacteria. Cefoxitin showed lower concentrations. Different pharmacokinetics for both of the test substances may be responsible for the different results.


[Encephalitis due to Naegleria and Acanthamoeba. Comparison of organisms and diseases (author's transl)]. Naegleria and Acanthamoeba are ubiquitous, free-living amoebas. Infections with Naegleria are acquired nasally by exposure to water and are characterized by an acute fulminant hemorrhagic necrotizing meningoencephalitis leading to death. Acanthamoeba-infections occur in chronically ill, debilitated individuals. A patchy chronic or subacute granulomatous encephalitis is produced by hematogenous spread of the amoebas. The histological or clinical diagnosis is not difficult.


Volatile fatty acids and aerobic flora in the gastrointestinal tract of mice under various conditions. Volatile fatty acids are reported to exert a repressive effect upon Enterobacteriaceae and Pseudomonas species in vitro and in vivo in young mice. The mean total volatile fatty acid concentration in the cecal samples of conventional mice fed ad libitum was 81.7 mumol/g (wet weight), which is antibacterial in vitro, and in the rectal samples it was 41.1 mumol/g (wet weight). The mean count of Enterobacteriaceae in the cecum was only 10(2)/g, whereas in the rectum it was 10(5)/g. Volatile fatty acid levels were influenced by food intake and increased to peak levels approximately 6 to 10 h after eating and then declined. In mice fasted for 17 h, the butyric acid concentration was considerably lower and the number of cecal samples positive for Enterobacteriaceae increased. When fasted for 4 days, mice had extremely low cecal and rectal volatile fatty acid concentrations and the Enterobacteriaceae and enterococci counts increased to mean of 2 x 10(6)/g and 3 x 10(6)/g, respectively, in the cecum and to means of 10(7) and 5 x 10(6)/g in the rectum. We conclude that volatile fatty acids are probably one of the many interference mechanisms which are involved with control of the levels of Enterobacteriaceae (and enterococci) in the large intestine of mice.


Effect of salts on water-insoluble glucan formation by glucosyltransferase of Streptococcus mutans. The formation of water-insoluble glucan by extracellular glucosyltransferase from Streptococcus mutans 6715 found to be greatly stimulated by various mono- or divalent cations. An enzyme preparation, obtained by ethanol fractionation, was able to catalyze the formation of water-insoluble glucan from sucrose in the presence of monovalent cations above 100mM or divalent cations above 20 mM at neutral pH. As the concentration of monovalent and divalent cations was reduced to below 10 mM and 1 mM, respectively, the formation of insoluble glucan decreased to a negligible amount. High concentrations of these cations were found to stimulate the formation of insoluble glucan in the following ways: (i) it increased the activity of total glucosyltransferase up to 1.6- and 2.7-fold in the absence and presence of a primer dextran, respectively, and (ii) it changed the formation of soluble glucan to insoluble. It was postulated that one of the essential factors for the formation of insoluble glucan would be to keep more than two water-soluble glucan chains close to enzyme aggregates and that such interaction could be enhanced by the presence of high cation concentrations.


Effect of bacteriocin production by Streptococcus mutans on the plaque of gnotobiotic rats. The effect of bacteriocins produced by strains of Streptococcus mutans on the microbial composition of dental plaque was studied in gnotobiotic rats. In one set of tests, using S. mutans strains T2 and SW31, rats were simultaneously infected with the bacteriocinogenic parent (bac(+)) and corresponding bac(-) mutants, and the relative proportions of each strain present in three different tooth sites were determined at various time intervals after inoculation. Some animals were monoinfected with bac(+) and bac(-) strains, and in vivo properties, such as plaque fermentation end products, amount of plaque formed, and cariogenicity, were measured. In another series of tests with strains T2 bac(+) and T2 bac(-), animals were sequentially inoculated with a time interval of 1 week. On simultaneous inoculation, the bac(-) mutant, which was sensitive in vitro to the bacteriocin produced by the parent, was totally unable to establish in a plaque consisting of the bac(+) strain even when the challenging dose was increased 100-fold. In contrast, the bac(+) strain was successful not only in invading an established bac(-) plaque but also in eventually almost eliminating the mutant. In monoinfected animals, there were no significant differences in plaque-forming ability, fermentation end products, or cariogenicity between the two strains. Also, bacteriocin activity was detected in plaque harboring the bac(+) but not the bac(-) strain. The results not only indicate that bacteriocin activity can occur in vivo but also suggest that bacteriocinogeny might confer an ecological advantage upon a strain attempting to establish itself in an ecosystem.


Properties of Streptococcus mutans grown in a synthetic medium: binding of glucosyltransferase and in vitro adherence, and binding of dextran/glucan and glycoprotein and agglutination. The influence of culture media on various properties of Streptococcus mutans was investigated. Strains of S. mutans (serotypes c, d, f, and g) were grown in a complex medium (Todd-Hewitt broth [THB]) or a synthetic medium (SYN). The SYN cells, in contrast to THB cells, did not bind extracellular glucosyltransferase and did not produce in vitro adherence. Both types of cells possessed constitutive levels of glucosyltransferase. B13 cells grown in SYN plus invertase-treated glucose possessed the same level of constitutive enzyme as THB cells. In contrast to THB cells, the SYN cells of seven serotype strains did not agglutinate upon the addition of high-molecular-weight dextran/glucan. Significant quantities of lower-molecular-weight (2 x 10(4) or 7 x 10(4)) dextran and B13 glucan were bound by SYN cells. SYN cells agglutinated weakly in anti-glucan serum (titers, 0 to 16), whereas THB cells possessed titers of 32 to 256. Evidence for the existence of a second binding site in agglutination which does not possess a glucan-like polymer has been obtained. B13 cells grown in invertase-treated THB agglutinated to the same degree as normal THB cells. The nature of this site is unknown. SYN cells possess the type-specific polysaccharide antigen. B13 cells did not bind from THB a glycoprotein which reacts with antisera to the A, B, or T blood group antigens or which allows agglutination upon the addition of dextran. The results demonstrate that S. mutans grown in a chemically defined medium possesse markedly different biochemical and biological activities than cells grown in a complex organic medium.


Purification and physicochemical and biological characterization of a staphylococcal pyrogenic exotoxin. A staphylococcal pyrogenic exotoxin was purified and characterized biochemically and biologically. The organism producing the toxin was a group I Staphylococcus aureus strain which was isolated from a vaginal infection of a patient with mucocutaneous lymph node syndrome (Kawasaki's disease). The possible association of the toxin with the disease syndrome is discussed. The toxin was purified from cell-free culture supernatant fluids by means of differential precipitation with ethanol and resolubilization in pyrogen-free distilled water followed by preparative thin-layer isoelectric focusing. The pyrogenic exotoxin produced fevers in both rabbits and mice and enhanced host susceptibility to lethal shock and myocardial and liver damage by endotoxin. Also, the toxin was a potent nonspecific lymphocyte mitogen, stimulating rabbit spleen cells and human cord blood lymphocytes to proliferate. The toxin migrated as a homogeneous protein when tested with sodium dodecyl sulfate-polyacrylamide gel electrophoresis (molecular weight, 12,000) and reisoelectric focusing (pI 5.3). Hyperimmune antisera raised against the purified toxin reacted with ethanol-precipitated toxin, using immunodiffusion to form a single precipitin arc. The toxin was distinguished from other staphylococcal toxins by a variety of methods. The amino acid composition was determined.


Inhibition of in vitro proliferative responses of human lymphocytes by rimantadine hydrochloride. Rimantadine hydrochloride (alpha-methyl-1-adamantanemethylamine hydrochloride) inhibits the in vitro proliferative response of human peripheral blood lymphocytes to mitogenic and antigenic stimulation. Addition of drug (25 micrograms/ml) at the initiation of 5-day cultures suppressed the responses to phytohemagglutinin, pokeweed mitogen, and concanavalin A by 25, 65, and 90%, respectively. Similarly, responses to streptokinase-streptodornase, tetanus toxoid, and A2/Aichi influenza vaccine were significantly inhibited at concentrations as low as 10 micrograms of rimantadine per ml. Viability studies on 5-day cultures using trypan blue exclusion showed no significant difference between drug-treated and untreated controls. Furthermore, addition of drug on day 3 of 8-day cultures, at a time when the majority of cells had undergone blastogenesis, greatly suppressed the responses to these mitogens. These studies suggest that, in addition to its antiviral action, rimantadine interferes with the generation of cellular immune responses.


Production of a mutagen from ponceau 3R by a human intestinal anaerobe. Ponceau 3R was reduced in vitro by Fusobacterium sp. 2, a human intestinal anaerobe, to a product which is mutagenic when metabolically activated by liver S9 preparations in the Salmonella/mammalian-microsome mutagenicity test. This mutagenic metabolite has been identified as 2,4,5-trimethylaniline.


Activation of the alternative complement pathway by L-phase variants of gram-positive bacteria. The present studies were performed to investigate the potential role of the alternative complement pathway in the host's defense against bacterial L-phase variants and to gain insight into the subcellular component of gram-positive bacteria responsible for activation of the alternative pathway. L-phase variants of Staphylococcus aureus and Streptococcus faecalis were able to activate the alternative pathway and consume C3 in C4-deficient guinea pig serum in amounts comparable to their respective bacterial-phase parent organisms. Activation of the complement system via the alternative pathway resulted in death of the L-phase variants. Membranes prepared from S. faecalis L-phase variants, by either osmotic lysis or mechanical disruption, retained their ability to activate the alternative pathway. Treatment of the membranes by three different methods (water washes, hot trichloroacetic acid, and cold trichloroacetic acid) resulted in a greatly diminished ability of the membranes to activate the alternative pathway. In addition, the extracts derived from the membranes by water washes and by cold-trichloroacetic acid treatment were able to activate the alternative pathway. These studies indicate that these L-phase variants can activate the alternative pathway and suggest that membrane-associated factors play a role in the alternative pathway activation by S. faecalis L-phase variants.


Local cholera immunity in mice: intestinal antitoxin-containing cells and their correlation with protective immunity. Methods for light and electron microscopic identification and characterization of intestinal cholera antitoxin-containing cells (ACC) using peroxidase-labeled immunoreagent are described and used to study the ACC response in mice after immunizations with cholera toxin. Specific ACC appeared in significant numbers after two oral immunizations and increased six- to eightfold with two additional oral boosters, whereas further oral immunizations caused no additional stimulation. Intravenous immunizations had to be repeated seven times before ACC could be detected. After two oral immunizations, most of the ACC were found in the proximal part of the small intestine and no ACC were seen in the colon. This uneven distribution of ACC within the small intestine was eliminated after four oral immunizations, when ACC could also be detected in the colon. The ACC response after four oral immunizations demonstrated a peak at 4 days with far fewer cells present at 2 and 7 days. Electron microscopic studies showed that the ACC were mature plasma cells with the staining localized to the endoplasmic reticulum. Regression analysis of the relationship between the number of ACC and the magnitude of protective immunity against intestinal challenge with cholera toxin indicates a highly significant correlation (r = 0.91, P less than 0.001).


Variable infection of Vero cells and homologous interference after co-cultivation with HeLa cells with persistent defective infection by Edmonston measles virus. The HeLa subline K11A-HG-1 (line of HeLa cells persistently infected with Edomonston measles virus but containing little or no transmissible infectious virus) was co-cultivated with Vero cells. Focal syncytia were formed containing measles antigen and accumulations of nucleocapsid-like structures with no detectable production of transmissible infectious virus or positive hemadsorption. The infection aborted between 2 and 3 weeks after preparation of co-cultures. Upon subculture of co-cultures, occasionally complete infections (progressive syncytial degeneration, hemadsorption, and production of transmissible infectious virus) appeared. A linear dose response curve for nontransmissible infection was obtained along with evidence that measles antigen had to be present on the surface of K11A-HG-1 cells for their infectivity for Vero cells. The basis for initiation of Vero cell infection by living K11A-HG-1 cells, but not by nonviable intact K11A-HG-1 cells killed by a virus-preserving technique, nor by disrupted K11A-HG-1 cells, is, at present, a matter of speculation. However, several lines of evidence were obtained which suggested that subsequent development of delayed variable transmissible Vero cell infection occurred because of a type of viral interference, including the presence of an inhibitor in K11A-HG-1 cultures, the bulk of which was cell-associated.


Ultrastructural changes of cultured human amnion cells by Clostridiu difficile toxin. The ultrastructure of the surface of primary human amnion monolayer cells undergoing cytopathology induced by Clostridium difficile toxin was examined by scanning electron microscopy. Our observations indicated that the type and distribution of cell surface projections were altered dramatically by this toxin. The patterns of such surface changes were specific for the two different types of cells found in this cell culture. Cells with demarcated borders showed rearrangement of microvilli into globular chains or ridges which lined up with the branching membrane. Cells without demarcated borders exhibited studlike microvilli, all arranged into ridges or globular chains. These changes were noted after 1 h of toxin exposure and persisted without further progression, in spite of continued toxin exposure, up to 48 h. These data indicate that C. difficile produces a cytolytic toxin and that scanning electron microscopy may be useful in determining toxin-cell interactions.


Cytotoxic enterotoxin produced by Aeromonas hydrophila: relationship of toxigenic isolates to diarrheal disease. Ninety-six Aeromonas hydrophila isolates were tested for cytotoxin and hemolysin production. Sixty-six (69%) of the isolates were both cytotoxic and hemolytic, whereas the rest produced neither cytotoxin nor hemolysin. No evidence of a separate cytotonic activity could be found in any of the isolates. Cytotoxin activity correlated with enterotoxic activity. Of four cytotoxin-producing strains tested in the isolated rabbit ileal loop, three were definitely positive and one was borderline, whereas two nontoxigenic strains were negative. Cytotoxin activity appeared to be a stable property and could not be assoicated with any common identified plasmid; only 10 of 21 cytotoxin-producing strains could be shown to have any plasmid by agarose gel electrophoresis. Cytotoxin production correlated with a positive lysine decarboxylase phenotype (98%) or a positive Voges-Proskauer phenotype (94%), compared to 27% lysine decarboxylase-positive and 23% Voges-Proskauer-positive, cytotoxin-negative isolates (P less than 0.001 for both). In fecal samples, cytotoxin production correlated with diarrheal disease; of 40 diarrheal isolates, 32 (80%) were toxigenic compared to 9 (41%) of 22 nondiarrheal isolates (P = 0.004). It appears that A. hydrophila can be a cause of diarrhea and that this enteropathogenic potential is mediated by a cytotoxic enterotoxin.


Experimental hematogenous Candida endophthalmitis: diagnostic approaches. Anterior chamber aspiration and vitreous aspiration were evaluated as diagnostic procedures for establishing a specific microbiological diagnosis in the rabbit model of hematogenous Candida endophthalmitis. Vitreous aspiration was the most successful procedure, confirming the diagnosis of hematogenous Candida endophthalmitis in 62% of eyes with documented intraocular infection. When animals with only the most severe clinical endophthalmitis were considered, vitreous aspiration confirmed the diagnosis in 89% of eyes evaluated. Vitreous aspiration correlated well with the extent of clinical endophthalmitis, as well as with postmortem, whole-vitreous cultures. Gram staining of the aspirate was additive to culture results in confirming the diagnosis. Anterior chamber aspiration was positive in only 1 of 58 eyes evaluated (1.7%). Additionally, muscle biospy was evaluated in this study as a tool for establishing a microbiological diagnosis in the rabbit model of disseminated candidiasis. Only 2 of 131 biospy specimens contained detectable Candida. Although vitreous aspiration may be associated with ocular complications, in certain clinical settings this procedure may be valuable in establishing the definitive microbiological diagnosis of hematogenous Candida endophthalmitis.


Purification and antigenic properties of intracellular invertase from Streptococcus mutans. Intracellular invertase from Streptococcus mutans GS5 was purified to near homogeneity by gel filtration and ion-exchange chromatography followed by preparative polyacrylamide gel electrophoresis. The invertase appeared to be composed of a single polypeptide chain with a molecular weight of 48,000. Extracellular invertase was identified in strain GS5 and was determined to have a molecular weight of 500,000. No antigenic relationship between these two forms of invertase was observed since antibody prepared against purified intracellular invertase neither affected extracellular invertase activity nor precipitated that enzyme on immunodiffusion. No antigenic relatedness between intracellular invertase and glucosyl- and fructosyl- transferases was detected since cross-reactivity with antibody prepared against either enzyme fraction was not observed after immunodiffusion. Using immunodiffusion and quantitative precipitin data, we examined the relationships of other S. mutans intracellular invertases to the serotype c enzyme. It appeared that the intracellular invertases from serotypes e, f, and g were structurally similar to the enzyme from serotype c, whereas the structure of invertases from serotypes a, b, and d appeared less similar to that of enzyme from serotype c.


Immunoregulatory responses in experimental disseminated histoplasmosis: lymphoid organ histopathology and serological studies. To study immunoregulatory mechanisms in systemic histoplasmosis, a highly reproducible model of infection was established in C3H/Anf mice. Intravenous inoculation of 6- to 8-week-old mice with from 5 x 10(5) to 10 x 10(5) cells of yeast-phase Histoplasma capsulatum strain G-217B produced disseminated infection that resolved over an 8-week period without therapeutic intervention. Serological studies demonstrated complement-fixing antibody production to yeast- and mycelial-phase antigens of H. capsulatum. Complement-fixing antibody to the former was detected at week 1, and it peaked at week 3 and declined thereafter. Complement-fixing antibody to mycelial-phase antigen(s) appeared later (week 3) and did not peak until week 18. Grossly, the spleens of infected mice were enlarged from three to four times normal size during peak infection, whereas the thymuses were markedly involuted. Conversely, at week 8 the average spleen size was considerably smaller and the thymic mass was increased relative to the mass at week 3. Histopathologically, the paracortical regions of lymph nodes and the white pulp (periarteriolar lymphocyte sheaths) and marginal zones of the spleen were heavily infiltrated by granulomata at week 1. By week 8, the infiltrates in these areas had largely resolved. Thymocytes were severely depleted from the cortical lobules of the thymus at week 1; however, thymic cellularity was restored by week 8. These reciprocal changes in cellularity of the thymus and spleen during active infection may be of importance with reference to the disturbances of immunoregulation that we have observed in Histoplasma-infected mice.


Isolation of mutants of Mycoplasma pneumoniae defective in hemadsorption. Mutants of Mycoplasma pneumoniae incapable of hemadsorption were isolated by means of chemical mutagenesis. These hemadsorption-negative mutants did not attach to hamster tracheal rings in vitro at as high a frequency as that exhibited by the wild-type parent strain.


Linkage analyses of extracellular glucans from Streptococcus sanguis and Streptococcus mitior. Similar alpha-(1-->6) linkage-rich, soluble, extracellular glucans have been isolated from six strains of two genetically distinct groups of Streptococcus sanguis and three strains of Streptococcus mitior.


Immunological relationship between delta-hemolysins of Staphylococcus aureus and coagulase-negative strains of staphylococci. Delta-hemolysin of coagulase-negative staphylococcal strains was immunologically identical to delta-hemolysin of Staphylococcus aureus (Newman) but distinct from delta-hemolysin from a canine strain of S. aureus.


In vivo and in vitro characterization of virulence-deficient mutants of Vibrio cholerae. In vitro and in vivo interactions between Vibrio cholerae and the infant mouse intestinal environment were examined by using a number of virulence-deficient mutants of strain CA401 which are unable to induce a typical diarrheal response. In vitro interactions with upper bowel sections were evaluated by determining percent association of radiolabeled organisms with sections. In vivo behavior was evaluated in the upper bowel early in infection with radiolabeled inocula. Ths relative degree of mechanical clearance was indicated by the percent recovery of input label. The relative degree of multiplication and killing was determined by changes in the specific activities (counts per minute per colony-forming unit) of inocula compared with recovered viable organisms. The results indicated that, whereas some virulence-deficient mutant classes exhibit net multiplication in the upper bowel, other classes show net killing in and accelerated clearance from the upper bowel. The in vitro association patterns failed to correlate with in vivo upper bowel recovery.


Serological studies on chemostat-grown cultures of Lactobacillus fermentum and Lactobacillus plantarum. Lactobacillus fermentum NCTC 6991 and Lactobacillus plantarum NCIB 7220 were grown in a chemostat in the diffusible fraction of complex medium at pH 6.0 with glucose limitation. Organisms grown at different dilution rates (D) were injected into rabbits, and the resultant antisera were examined for reactivity with antigens previously isolated from batch-grown organisms. For L. fermentum, antisera obtained on injecting cells grown at D = 0.5 h-1 contained a significantly higher level of antibody reacting with lipoteichoic acid and a lower level of antibody reacting with wall polysaccharide than did antisera obtained with slower-growing cells (D = 0.05 and 0.033). Antibodies to the cell wall polysaccharide were alpha-D-glucosyl specific and cross-reacted with dextran and alpha-D-glucosyl ribitol teichoic acid from L. plantarum. The immunogenicity of the ribitol teichoic acid and lipoteichoic acid components of L. plantarum was not influenced by injecting organisms grown at different rates. However, chemical and serological studies indicate that growth of L. plantarum in the diffusible fraction of complex medium results in a wall teichoic acid of lower glucose substitution. This apparently influences the specificity of the resultant antibodies so that some sera react much less with glucosyl-substituted lipoteichoic acid and dextran.


Influence of malaria infection on the elaboration of soluble mediators by adherent mononuclear cells. MALARIA RESULTS IN TWO SEEMINGLY PARADOXICAL PERTURBATIONS OF THE IMMUNE RESPONSE: polyclonal B-cell activation and immunosuppression. To determine what immunoregulatory role mediators secreted by adherent cells might play in these alterations, we cultured adherent cells from uninfected mice and from mice at different times during infection with Plasmodium berghei or P. yoelii. Culture supernatants obtained from these cells were tested for their ability to enhance the in vitro proliferative responses of thymocytes to suboptimal concentrations of concanavalin A or to inhibit the mitogen-stimulated proliferation of normal spleen cells. Supernatants obtained from adherent cells of mice early in infection (days 1 to 3) contained significantly elevated levels of enhancing activity which on Bio-Gel P-100 chromatography resembled lymphocyte-activating factor. Later in infection (days 4 and 5), these supernatants contained inhibitory activity. Normal adherent cells, when cocultivated in vitro with parasitized erythrocytes, ingested parasite debris and were stimulated to produce the enhancing factor. At high parasite/adherent-cell ratios, cells elaborated an inhibitory factor. These findings suggest that during malaria, adherent cells are converted from a nonspecific helper role to a nonspecific suppressor role. This modulation in function may be due to the direct interaction between adherent cells and parasitized erythrocytes.


Immunoglobulin G from subacute sclerosing panencephalitis brain as an immunological reagent. Proteins were extracted at pH 2.5 from carefully washed brain homogenates of patients with subacute sclerosing panencephalitis. Immunoglobulin G was isolated from the extracts by gel filtration and immunoadsorbent chromatography and was 125I radioiodinated. The labeled immunoglobulin G was reacted with purified measles virus antigens by crossed immunoelectrophoresis and utilized to identify antigens in measles-infected cell cultures and hamster brain tissue subjected to treatment corresponding to that used for elution of immunoglobulin G. The method described is useful in identifying the specific nature of antibodies as well as antigens in tissue-associated immune complexes.


Immune electron microscopy of cross-reactions between Mycoplasma pneumoniae and human erythrocytes. Respiratory infection with Mycoplasma pneumoniae evokes immunoglobulin M autoantibody which agglutinates human erythrocytes at 4 degrees C (cold agglutinin) and is specific for I antigen. Cross-reactions between surface antigens of M. pneumoniae and human erythrocytes, previously examined by serological analysis, were examined by transmission and scanning electron microscopy. Ferritin-labeled human antimycoplasmal and rabbit antisera to erythrocyte membrane components reacted with antigens on the surface of both M. pneumoniae and erythrocytes. Adsorption of human erythrocytes to M. pneumoniae was blocked by the same antisera without ferritin label. It is proposed that the cross-reactive specificity lies in peripheral areas of the mycoplasmal cell, probably in a surface carbohydrate which has antigenic identity with erythrocyte glycoprotein.


Biotyping of Chlamydia psittaci based on inclusion morphology and response to diethylaminoethyl-dextran and cycloheximide. Strains of Chlamydia psittaci from cattle, sheep, pigs, mice, guinea pigs, rabbits, cats, and parrots were subdivided based on their biological characteristics. Chlamydiae grown in the yolk sac of chicken embryos were used to infect L cell monolayers. The host cells were infected without further treatment or treated with diethylaminoethyl-dextran, cycloheximide, or both. The following criteria were used for biotyping the strains: the morphology of the inclusions and time after infection at which they appeared, the effect of chlamydial multiplication on the host cell cytoskeleton, and the change in the number of cells infected in response to diethylaminoethyl-dextran and cycloheximide. These properties were determined for 29 strains of C. psittaci. Based on the results, the strains were placed into eight biotypes.


Local antibody response in chickens: analysis of antibody synthesis to Newcastle disease virus by solid-phase radioimmunoassay and immunofluorescence with class-specific antibody for chicken immunoglobulins. The antibody response to Newcastle disease virus was monitored in the sera and salivas of adult chickens immunized by two methods: (i) combined intratracheal-intranasal vaccination followed by intratracheal revaccination or (ii) intramuscular vaccination followed by intratracheal revaccination. By solid-phase radioimmunoassay, only immunoglobulin G (IgG) and IgA antibodies to Newcastle disease virus were detected in the salivas, whereas IgA and IgM antibodies were present in egg whites. The first method produced the highest antibody levels in both serum and saliva and, in addition, prevented detectable virus multiplication in the respiratory tracts upon revaccination 4 weeks later. Plasma cells of all three classes were distributed throughout the tissues lining the oral cavities. The highest densities of plasma cells were in the Harderian glands; IgG was the predominant class, whereas IgA and IgM plasma cells were present in almost equal but lower numbers. The Harderian plasma cells were the most likely source of the antibody found in saliva.


Enhancement of human neutrophil bactericidal activity by chemotactic factors. Neutrophils are important effector cells in the defense against microorganisms. They migrate into infected sites and then phagocytose and kill bacteria. Chemotactic factors may be important for initiating neutrophil migration. We investigated whether chemotactic factors might also influence an event subsequent to chemotaxis, namely bacterial killing. It was found that preincubation (20 min at 37 degrees C) of human leukocytes with chemotactic substances such as zymosan-activated serum, a C5a-containing fraction of zymosan-activated serum, N-formyl methionyl phenylalanine or N-formyl methionyl-leucine-phenylalanine, enhanced leukocyte killing of Staphylococcus aureus, Escherichia coli, and Streptococcus faecalis in a dose-dependent fashion. The concentration of chemotactic factor required to enhance killing was similar to that required to induce neutrophil chemotaxis. In addition, zymosan-activated serum, C5a fraction, and the two N-formyl methionyl peptides increased the hexose monophosphate shunt activity of resting and phagocytosing neutrophils by two- to threefold. In contrast, bacterial killing by sodium azide-treated neutrophils and neutrophils from a patient with chronic granulomatous disease was not increased by any chemotactic factor. These findings suggest that chemotactic factors stimulate neutrophil oxygen-dependent microbicidal pathways. These observations illustrate another important contribution of biologically active molecules to effector cell function and host defense.


Stimulation of the reticuloendothelial system of mice by muramyl dipeptide. Muramyl dipeptide (MDP), a part of bacterial cell wall peptidoglycans, was active as adjuvant and stimulated the reticuloendothelial system (RES) of mice to increase its phagocytic function. A series of analogs of MDP was tested for their adjuvant activity and RES-stimulating activity. Sex differences were observed in the adjuvant activity and RES-stimulating capacity of some MDP analogs. A stereochemically highly specific structure required for MDP to exert adjuvant activity was also required for its RES-stimulating activity. Based on this close correlation among the structure, adjuvant activity, and RES-stimulating capacity of MDP, we infer that macrophages may play an important role in the expression of adjuvant activity of MDP.


Inhibition of macrophage migration by muramyl peptides. In the capillary tube migration system a synthetic muramyl dipeptide (MDP; N-acetylmuramyl-L-alanyl-D-isoglutamine), a part of bacterial cell wall peptidoglycans, inhibited the migration of peritoneal exudate macrophages from normal guinea pigs or rats. The migration inhibition was also caused by some MDP-containing peptidoglycan fragments from cell walls of Lactobacillus plantarum and Staphylococcus epidermidis. The migration inhibition could not be explained on the basis of macrophage migration inhibitory factor. A stereochemically highly specific structure of MDP required for its adjuvant activity was also required for the macrophage migration inhibition. These findings suggest that MDP and MDP-containing cell wall fragments may activate macrophages and that this activation may be important in the exertion of their adjuvant activity.


Adjuvant activities in production of reaginic antibody by bacterial cell wall peptidoglycan or synthetic N-acetylmuramyl dipeptides in mice. This paper is concerned with the adjuvant activity in stimulatory immunoglobulin E production against ovalbumin (OA) by bacterial cell walls, cell wall peptidoglycan (PG), and their PG fragments and synthetic N-acetylmuramyl (MurNAc) dipeptides in A/J mice. A PG isolated from Streptococcus pyogenes, PG subunit polymer and dimer obtained from Staphylococcus epidermidis, and water-soluble fragments of cell walls or PG prepared from Nocardia corynebacteriodes and Streptomyces gardneri were found to enhance both the primary and secondary responses of anti-OA immunoglobulin E antibody production. It was suggested that the PG portion, either intact or highly degraded, was capable of enhancing the immunoglobulin E antibody production, and there was no need for the non-PG moiety or intactness of PG structure for the adjuvant activity. This finding was confirmed and extended by the use of synthetic MurNAc dipeptides. Among eight MurNAc dipeptides tested, MurNAc-l-Ala-d-isoGln, MurNAc-l-Ala-d-Gln, MurNAc-l-Ala-d-Glu, and MurNAc-l-Ser-d-isoGln were found active as an adjuvant in the stimulation of the primary and secondary reaginic anti-OA antibody production in a similar way to the cell wall PG and their fragments. None of the synthetic MurNAc-l-Ala-l-isoGln, MurNAc-l-Ala-l-Gln, MurNAc-l-Ala-l-Glu, and MurNAc-l-Ala-d-isoAsn, on the other hand, stimulated the anti-OA immunoglobulin E antibody production in either primary or secondary response, indicating the importance for the adjuvancy in immunoglobulin E production of the configuration of the glutamic acid residues adjacent to the l-Ala (or l-Ser) in muramyl dipeptides.


Hepatitis B e-antigen and its correlation with other serological markers in chimpanzees. Three chimpanzees experimentally infected with hepatitis B virus, and another three chimpanzees that were hepatitis surface antigen carriers, were studied for the presence of viral antigens and humoral immune responses. Quantitative analyses of hepatitis B surface and e-antigens in sequential serum samples at early acute stages revealed cyclic oscillations of these two antigens following a synchronous pattern. Similar analyses of anti-e-antigen and anti-hepatitis B core antigen antibodies from the three experimentally infected primates indicated that peak titers of these two antibodies occurred as surface antigen decreased to undetectable levels. Of the three surface-antigen carriers, two were positive for e-antigen and one was positive for e-antigen antibody for the entire course of surveillance (8, 9, and 22 months, respectively).


Suppression of local intestinal immunoglobulin A immune response to cholera toxin by subcutaneous administration of cholera toxoids. Cholera toxin has been shown to modulate immune responses, generally producing enhancement when administered simultaneously with antigen and suppression when administered a day or more earlier. In a previous study using chronically isolated ileal loops in rabbits, we found that two subcutaneous (s.c.) "priming" and "boosting" doses of biologically active cholera toxin suppressed the local intestinal immunoglobulin A response to intraloop doses of cholera toxin. In the study reported here, two different biologically inactive but antigenically intact cholera toxoids, glutaraldehyde toxoid and choleragenoid, where administered s.c. by the same immunization schedule as for toxin in the earlier experiment. Suppression of local immune response to intraloop cholera toxin as compared with animals receiving no s.c. inoculations was again found. The results suggest that in this model suppression was immunological (mediated by an immunological mechanism) rather than toxigenic (mediated by biological activity of cholera toxin). In addition, the occurrence of suppression of local intestinal immune response after systemic immunization suggests that suboptimal protection against enteric infections could occur after s.c. vaccination.


Abortion in laboratory animals induced by Moraxella bovis. Pregnant mice, guinea pigs, rats, and rabbits responded to injections of Moraxella bovis strain EPP-63(300) with abortion, death, embryo resorption, and production of small litters. The nature of response appeared to depend primarily on the number of viable cells injected and to some extent on the species of animal and stage of pregnancy. Intraperitoneal injection of mice with 3 X 10(5) viable cells induced 100% abortion and no deaths. Embryo resorption and smaller litters were induced with injection of lower doses of M. bovis. None of the rats used in this study aborted; however, resorption rates were higher in rats than mice. Bacteria-free filtrates prepared from aqueous extracts of blood agar on which M. bovis had been grown induced abortion in mice and guinea pigs, at a rate similar to that caused by viable cells. The lyophilized filtrate could be diluted to produce 100% abortion with no death of injected mice. Abortion-inducing, nonlethal doses of M. bovis and lyophilized filtrates appeared to have no effect on nonpregnant female or male mice.


Depressed mitogen responsiveness of lymphocytes at skin temperature. The responsiveness of murine lymphocytes and human peripheral blood lymphocytes to phytohemagglutinin, concanavalin A, pokeweed mitogen, and endotoxin was tested in vitro at 32, 35, and 37 degrees C. The responses at 32 degrees C were delayed and often depressed. Mouse cells responded equally well at 35 and 37 degrees C. Human lymphocytes often responded more rapidly at 37 than at 35 degrees C. Since skin temperature, particularly that of the distal extremities, is usually 32 degrees C or less, a relative deficiency in cell-mediated immunity may exist in these sites. This may be part of the reason for the usual localization of certain infections, such as sporotrichosis, to these coller areas.


Two-dimensional gel electrophoretic comparison of proteins from virulent and avirulent strains of Mycoplasma pneumoniae. The protein composition of the virulent M129 strain of Mycoplasma pneumoniae was compared to that of its homologous avirulent strain by the use of standard one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Forty-nine individual M. pneumoniae cell proteins were resolved by this method, and the virulent strain was shown to possess a single high-molecular-weight protein not present in avirulent cells. Variability in the resolution of this particular protein in one-dimensional gels prompted the application of two-dimensional gel electrophoresis to the analysis of M. pneumoniae cell proteins. The sequential use of isoelectric focusing in the first dimension and sodium dodecyl sulfate-polyacrylamide gel electrophoresis in the second dimension permitted the resolution of a least 142 individual M. pneumoniae cell proteins. Application of nonequilibrium pH gradient electrophoresis in the first dimension achieved the resolution of at least 20 additional basic proteins. Three proteins which are synthesized only by cells of the virulent strain, and not by the homologous avirulent strain, were identified by these two-dimensional gel electrophoresis techniques.


Streptococcal infection of endocardial and other intravascular vegetations in rabbits: natural history and effect of dexamethasone. Experiments were designed to study the natural history of infection in different parts of the vascular system. Sterile vegetations were produced in rabbits by placing catheters in the inferior vena cava, tricuspid or aortic valves, and thoracic or abdominal aorta and then were infected by the intravenous inoculation of Streptococcus sanguis. At 1 day after bacterial challenge, all VEGS were infected, mean bacterial densities being highest in the VEGS of the aortic and tricuspid valves. By 14 days, there was a significant decrease in the mean bacterial density in all VEGS except for the aortic valve: the VEGS of the inferior vena cava and abdominal aorta were sterile, as were half of those of the thoracic aorta. There were no deaths except for animals with aortic valve infection. Dexamethasone inhibited the sterilization of the thoracic aorta VEGS, but was without effect on aortic valve VEGS, 5 mm distant. Sterilization of tricuspid valve VEGS after catheter removal was also inhibited by dexamethasone. Thus, there are host defense mechanisms which lead to the sterilization of infections everywhere in the vascular system except in the left side of the heart, and these mechanisms, as yet undefined, are inhibited by dexamethasone.


Suppression of tuberculin hypersensitivity during influenza infection in mice. Tuberculin hypersensitivity was examined during acute influenza infection in mice. Employing the footpad test as a measure of delayed-type hypersensitivity, it was noted that tuberculin hypersensitivity was suppressed temporarily beginning on day 3 and continuing through days 10 to 16 following intranasal infection with influenza A/PR8. These changes occurred at a time when influenza virus was replicating in lung tissue. Suppression of footpad swelling was not detected when mice were administered live virus intravenously, were given Formalin-inactivated virus intranasally, or were immunized against influenza before intranasal infection. Transient reduction of total circulating lymphocytes also occurred during influenza infection but did not correlate with the duration of footpad suppression. Because this model system reproduces many of the alterations in immunological function reported to occur during influenza infection in humans, it should provide a useful tool for investigating mechanisms of influenza-induced immunosuppression.


Electrophoresis of buffalo (bos bubalis) serum proteins including immunoglobulins. Antigenic components of buffalo (Bos bubalis) serum, which were also components of buffalo colostrum, seminal plasma, milk whey, saliva, and tears, were investigated by the ager gel diffusion test and immunoelectrophoresis. Immunoglobulins of buffalo serum were identified by immunoelectrophoresis employing rabbit-anti-buffalo serum and rabbit-anti-buffalo gamma-globulin. Based on immunoelectrophoretic patterns immunoglobulin G (IgG), IgGA, and IgM were detected both in the serum and colostrum of buffaloes. Tears contained both IgG and IgM. Cross-reactions of buffalo serum with seminal plasma, saliva, and milk whey were observed only in the IgG region. By polyacrylamide gel electrophoresis, lipoprotein (5.2% +/- 0.41), IgM (11.4% +/- 3.1), IgG (9.4% +/- 0.98), haptoglobin 21.8% +/- 3.73), transferrin (10.4% +/- 2.15), ceruloplasmin (7.8% +/- 1.3), postalbumin (20.8% +/- 2.09), and albumin (13.7% +/- 0.75) were identified provisionally.


Mechanism of lipopolysaccharide-induced tumor necrosis: requirement for lipopolysaccharide-sensitive lymphoreticular cells. Lipopolysaccharide (LPS) induces rapid necrosis of intradermal fibrosarcomas in mice. The mechanism(s) by which LPS produces tumor necrosis has been investigated using histocompatible LPS-sensitive (C3H/HeN) and LPS-resistant (C3H/HeJ) mouse strains. C3H/HeN- or C3H/HeJ-derived fibrosarcomas were necrotized by LPS when they were grafted onto C3H/HeN mice but were not affected when growing on C3H/HeJ mice, indicating that LPS does not act directly on the tumor itself. In contrast, lethally X-irradiated C3H/HeJ mice exhibit necrosis of their tumors when reconstituted with C3H/HeN bone marrow cells, whereas C3H/HeN mice no longer exert LPS-induced tumor necrosis after the adoptive transfer of C3H/HeJ bone marrow cells. These findings clearly indicate that LPS produces necrosis of tumors by activating host lymphoreticular cells.


Effect of sulfolipid I on trehalose-6,6'-dimycolate (cord factor) toxicity and antitumor activity. Emulsified mixtures of sulfolipid I and trehalose-6,6'-dimycolate (cord factor) had tumor-regressive activity comparable to, but less toxic than, emulsified trehalose-6,6'-dimycolate alone.


Differential effects of interferon on the MHC expression of human lymphocytes. Enhanced expression of HLA without effect on Ia. HLA antigens are the principal serologically detectable products of the major histocompatibility complex (MHC), and they function as targets for antibody-mediated and cell-mediated cytolysis. Anti-HLA sera were used in a quantitative absorption procedure to study the effect of interferon (IF) treatment on HLA expression. This study was undertaken since IF has been shown to play an important role in the regulation of cell division and function. We found that IF enhanced the expression of HLA antigens on human peripheral blood lymphocytes by 8-fold. This increase in HLA expression was specific for both the HLA-A and HLA-B regions of the MHC. There was no increase in the expression of the Ia region after IF treatment as opposed to the HLA region. Since IF is an antiviral agent and it also enhances the expression the HLA-A and HLA-B regions, it may be involved in the elimination of virus-infected cells by A and B identical effector cells.


Paul-Bunnell antigen in malignancies and rheumatoid arthritis. By means of agglutination inhibition test, Paul-Bunnell (P-B) antigen was demonstrated in sera of patients with cancer (5%), lymphomas or leukemias (12.5%) and rheumatoid arthritis (RA) (1.4%) as well as in synovial fluids from RA patients (27%). In contrast, none of 124 normal human sera gave positive results. P-B antigen was also demonstrated in 3 of 12 perchloric acid extracts of cancer tissues and 2 of 11 saline extracts obtained at 100 degrees C from spleens of lymphoma-leukemia patients. Extracts of apparently normal tissues from 16 individuals gave negative results.


Effect of cyclophosphamide on the cellular infiltrate in experimental allergic contact dermatitis. The influence of cyclophosphamide on contact allergy was studied by a method based on the count of cells infiltrating epicutaneous test sites in sensitized guinea pigs. Cyclophosphamide was injected 72 h before sensitization with dinitrochlorobenzene (DNCB). On testing 2 weeks later, marked mononuclear cell infiltration and dramatic increase in the count of basophil polymorphonuclear cells in the upper corium were found, compared to the control group. Cell infiltration in tests performed 1 and 3 weeks after sensitization differed from the reaction after 2 weeks. When cyclophosphamide was given 72 h before testing done 2 weeks after sensitization, both mononuclear and polymorphonuclear counts were found to be greatly reduced. This in vivo method of assessing the degree and composition of cellular infiltration in a contact allergic reaction after different periods of time throws new light on the influence of drugs on a cell-mediated immune response and also the variation in response with time.


Complement activation by rat platelets: its significance to inflammation in the rat. Generation of chemotactic activity in rat plasma or serum by aggregated rat platelets or by an extract of lysed platelets has been demonstrated. Chemotactic activity was not generated from serum either in the absence of an intact complement system or by platelet lysate pre-incubated with soya bean trypsin inhibitor. The significance of these observations is discussed in relation to some recent work on the role of platelets in inflammation.


Serum factors affecting the incorporation of [3H]thymidine by lymphocytes stimulated by antigen. IV. Comparison of enhancement by heated (56 degrees C) serum and by 2-mercaptoethanol. Serum preheated to 56 degrees C and 2-mercaptoethanol (ME) both enhance the stimulation (%) by antigen of [3H]thymidine incorporation by lymph node cells from immunized rabbits. The enhancements show several similar features suggesting a common mechanism. Both enhancements show (i) non-specific components, affecting [3H]thymidine incorporation in control and antigen-treated cultures proportionately, and (ii) antigen-specific components, having a greater proportionate effect on antigen-treated cultures than on control cultures. The non-specific components, in combination, are approximately additive. The specific components, in combination, are non-additive (mutually exclusive). With both preheated serum and ME the specific component of the enhancement is most evident late in the culture period and is greater at high antigen concentrations. The ME concentration needed for optimum enhancement is reduced if the serum concentration is reduced or if serum is preheated.


Antibody formation in experimental amyloidosis. In endotoxin-induced amyloidosis of mice it was found that all mouse strains formed antibodies specific for the type of endotoxin injected. In casein-induced amyloidosis, the animals form antibodies to certain components of the casein and most likely to whatever contaminating endotoxin is present. The antibodies formed seemed to be primarily of the IgG 1 subclass. Their appearance in high titer shows a good correlation with the onset of amyloid formation especially in mice given casein. Failure to form amyloid in these experimental models does not seem to be related to failure of antibody formation but may be explained by more effective clearance of immune complexes or other such factors.


High oxygen pressure inhibits DNA synthesis in mitogen-activated lymphocytes. Peripheral sheep lymphocytes, exposed to the mitogenic lectin phytohemagglutinin do not incorporate radiolabelled thymidine in cultures with a gas phase high in oxygen. Cells kept under nitrogen incorporate thymidine to the same extent or better than cells grown under air. The inhibition is proportional to the oxygen content of the gas phase. The inhibition is relieved if oxygen is replaced by air after some time but the onset of DNA synthesis is delayed correspondingly. It is suggested that oxygen may have a regulatory function on the onset of DNA synthesis.


Stability of peritoneal urea clearance in continuous ambulatory peritoneal dialysis (18 months experience with CAPD). A group of 13 patients on Continous Ambulatory Peritoneal Dialysis up to 18 months was studied. During the study period, Peritoneal Urea Clearance was stable. In all but 2 cases peritonitis did not depress the efficiency in clearing urea. When Peritoneal Urea Clearance was corrected for 1.73 m2 surface area, there was on the average an 11.8% increase over not corrected clearance rates. This increase was statistically significant.


A two and one half year clinical experience with a mechanical left ventricular assist pump in the treatment of profound postoperative heart failure. A sac-type paracorporeal left ventricular assist pump was implanted in 9 patients suffering from profound left ventricular failure following open heart surgery. Two patients had good recovery of left ventricular function and were weaned from assist. One survived 14 months postoperatively. Primary causes of death among the remaining patients were (1) primary right ventricular failure, (2) assist pump inlet cannula obstruction and (3) excessive bleeding. The strategies developed or proposed to deal with these problems include, respectively: (1) biventricular assist for patients with concomitant right ventricular failure; (2) cannulation of the left atrium to avoid inlet cannula obstruction by the left ventricular wall and intraventricular septum; (3) more rapid selection of left assist candidates and more rapid implantation techniques to reduce the coagulopathy associated with prolonged cardiopulmonary bypass, and an effective autotransfusion system to augment blood replacement.


Urea and ammonium removal based on alkalinization and removal of free ammonia. A novel approach is reported for the removal of ammonium formed from the conversion of urea by urease. By alkalinization, ammonium is converted into free ammonia. Free ammonia can then be very easily removed by a number of approaches: as gaseous ammonia by air bubbling, oxygenator, or air ventilation; by adsorbent for free ammonia; or other approaches.


In-vitro assessment of the removal of phenols by ACAC hemoperfusion. The in-vitro removal of free phenols by means of perfusion over albumin coated cellulose nitrate microencapsulated activated charcoal (ACAC) was studied. The phenols tested were in the form of a standard solution comprised of phenol, p-cresol, and p-hydroxyphenylacetic acid in a ratio of 1:5:14. Four different activated charcoals were tested for their adsorption capacity. Further studies were conducted to determine; the effect of microencapsulation and albumin coating of the activated charcoal on the adsorption of phenols; the adsorption isotherm of phenols; the capacity; and the clearance values. The results obtained suggest that ACAC used in the management of patients with uremia or hepatic coma is extremely efficient in removing phenols in aqueous solution in the in-vitro situation.


Methylation of DNA in target and non-target organs of the rat with methylbenzylnitrosamine and dimethylnitrosamine. The sites of labelling of DNA with [14C]methyl groups from methylbenzylnitrosamine and dimethylnitrosamine were studied in rat oesophageal epithelium and liver. All four combinations of tissue and carcinogen were studied. Tissues were labelled in vitro and the DNA contained therein purified and hydrolysed (pH 1, 37 degrees C) to free purines and apurinic acid. Quantitative analysis was performed with the aid of thin-layer chromatography. The apurinic acid and 7-methylguanine fractions were found to be extensively labelled. Smaller amounts of radioactivity were found in O6-methylguanine and some of the methylated adenines. The same carcinogen produced different patterns of labelling is oesophageal and liver DNA. The proportion of O6-methylguanine to 7-methylguanine was higher when the methylating agent was a carcinogen specific for the organ.


Transplanted cultured cells from pregnancy-dependent mammary tumors have a heterogenous developmental potential. Cultured cells derived from pregnancy-dependent mouse mammary tumors (plaques) when transplanted into the gland-free mammary fat pad display a heterogeneous developmental potential. They not only gave rise to plaques but also produced normal mammary gland, hyperplastic alvoelar nodules (HAN), and autonomous mammary tumors.


Volar melanotic macules.

Volar melanotic macules. Asymptomatic light brown or tannish-gray macules are seen on palms and/or soles of black patients, and occasionally on the volar surfaces of whites. They may be mistaken for lesions of secondary syphilis or other postinflammatory hyperpigmented dermatoses. Histologic examination of 14 specimens obtained at necropsy showed purely epidermal hyperpigmentation of all epidermal layers; or (3) melanin restricted to large dendritic melanocytes without appreciable transfer to keratinocytes. The number of melanocytes was not significantly increased, and with one exception, there were no nevus cells. These volar melanotic macules have close clinical and histologic resemblance to melanotic macules observed occasionally on the vermillion area of the lips.


Systemic absorption of topical salicylic acid. The systemic absorption of salicylic acid in humans following topical application in either hydrophilic ointment or polyethylene glycol 400 vehicle was studied. Drug absorption was minimal following application to intact skin; however, measurable levels (8 mg/dl) were attained when the stratum corneum was removed prior to application of the drug in hydrophilic ointment. A one-compartment open model with first order absorption and elimination processes was fitted to the plasma salicylate concentrations as a function of time. Computer simulations predict that plasma salicylate levels associated with toxicity in some patients may be present after repetitive application of the drug in hydrophilic ointment.


Demographic study of head lice infestations in Sacramento County school children. Head lice infestations have become a problem in school children in Sacramento County. An open study was conducted to determine how widespread head lice infestations were among preschool and elementary school children in Sacramento County and to see how effective a newly marked ever-the-counter pediculicide containing 0.3% pyrethrins synergized by 3.0% piperonyl butoxide was. The public health nurse for this study visited the homes of all reported patients infested with head lice. She verified and provided the pyrethrins-based treatment for 248 children, aged 6 months to 12 years. The children were Caucasian, Mexican-American, black and other ethnic backgrounds and from low, middle and upper income levels. All the patients were successfully treated, and the school children were able to return to their classs in a short time.


Theoretical conformational analysis of oxytocin molecule. The total semi-empirical conformational analysis of the oxytocin molecule has been carried out. It has been revealed the two main types of stable structures of cyclic moiety backbone and the great lability of the tail. The optimal spacing of cyclic moiety side chains has been found for every backbone structure. The calculation results are in good agreement with the data of physico-chemical investigations. Among the set of stable molecule structures reported in the present study are structures with beta-turn conformation of the cyclic moiety backbone and without closer spacing of the cyclic moiety and the tail, as well as structures with closely spaced N- and C-terminal parts which, however, lack beta-turn in the cyclic moiety.


X-ray study on homo-oligopeptides t-BOC-(D-Ala)7-OMe and HCl.H(D-Ala)7-OMe. Observations of extended peptide chains, whose direction is perpendicular to the fiber axis (cross-beta-structures) have hitherto been confined to fibrous proteins and to some synthetic polydisperse polypeptides of rather low molecular weight. This structure has now been found in some monodisperse linear homo-oligopeptides with aliphatic hydrocarbon side chains. X-ray diffraction photographs of t-Boc-(D-Ala)7-OMe and HCl.H(D-Ala)7-OMe show characteristic reflections for this form. In addition, the good orientation of suitably prepared specimens has enabled a fairly complete determination of the unit cell of the heptapeptides to be made. t-Boc-(D-Ala)7-OMe and HCl.H(D-Ala)7-OMe molecules are packed in a monoclinic lattice with a = 4.80, b = 34.5, c = 5.82 A, beta = 65.6 degrees and a = 4.80, b = 29.5, c = 5.82 A, beta = 65.6 degrees, respectively. It has not been possible to establish whether the arrangement of the chains within the sheets is parallel or antiparallel.


A series of lysyldipeptide derivatives for racemization studies in peptide synthesis. The series of diastereomeric peptide derivatives N-benzoyl-D,L-X-N epsilon-benzyloxycarbonyl-L-lysine methyl ester where X = alanyl, valyl, leucyl, phenylalanyl and isoleucyl are submitted as model systems for studying racemization in peptide synthesis. The diastereomers can be analyzed by quantitation of the separated ester methyl proton peaks of their nuclear magnetic resonance spectra. Data on the tendency to racemize of the different residues are presented. In polar solvents, valyl and isoleucyl residues racemize more readily than the other residues.


Isolation and physical characterization of bovine lens crystallins. The soluble proteins from bovine lens homogenate were separated on Sepharose CL-6B (2 X 200 cm) in 0.05 M tris-NaHSO3 pH 8.2 buffer containing 20 mM EDTA. Five sharp and defined fractions (HM alpha, alpha, beta H, beta L, gamma) were obtained. Each crystallin fraction was further purified by rechromatography on the same column. Each protein fraction was pure as judged by ultracentrifugation and SDS-gel electrophoresis. The molecular weights of the five fractions were 3.04 x 10(6), 5.83 x 10(5), 1.58 x 10(5) , 4.59 x 10(4), 2.14 x 10(4) as determined from sedimentation coefficient and intrinsic viscosity data by Scheraga-Mandelkern equation, which was in close agreement with that obtained by gel filtration. The polypeptide composition of crystallins as determined by SDS-gel electrophoresis revealed one band for high molecular weight alpha (HM alpha) and alpha, three for beta H, two for beta L and one for gamma. The gross CD patterns of crystallins were about the same in the peptide region (200 nm similar to or approximately 250 nm) with a minimum centered at about 217 nm, indicative of a beta-sheet structure in all crystallins. The [theta] values at 217 nm ranged from --1700 to --3700 degrees cm2 per decimole. The CD spectra of these crystallins in the aromatic region (250 nm similar to or approximately 300 nm) were different, reflecting the different contributions of aromatic amino acids to the tertiary structure of crystallins.


Use of cyclopentyl ester protection for aspartic acid to reduce base catalyzed succinimide formation in solid-phase peptide synthesis. A study was conducted to determine the effect of amino acid sequence and aspartyl protecting group on the rate of base catalyzed succinimide formation in the solid-phase synthesis of aspartyl peptides. The peptides H-Ala-Asp-Gly-Phe-OH and H-Ala-Asp-Leu-Phe-OH were synthesized by the solid-phase method with cyclopentyl or benzyl protection for the beta-carboxyl of aspartic acid. The results showed that the cyclopentyl ester was notably less susceptible to succinimide formation by treatment with tertiary amine than was the benzyl ester, and that the difference could have significant consequences for the synthesis of the large peptides which contain reactive sequences such as Asp-Gly.


Synthesis and some pharmacological properties of [Glu(OMe)4]oxytocin and [Mpr1, Glu(OMe)4]oxytocin. [Glu(OMe)4]oxytocin (XVI) and [Mpr1, Glu(OMe)4]oxytocin (XVII) bearing a methyl ester group in place of the carboxamide group in position 4 of oxytocin were synthesized by (3 + 6) segment condensation using the S-trityl group for the protection of the cysteine side chains. Analogue XVI exhibited 10.5 U/mg in vitro uterotonic, and 42 U/mg avian vasodepressor, activity, and analogue XVII 21.4 U/mg and 82 U/mg of the respective activities. Both compounds showed no response in the rat pressor assay.