Nickel, cobalt, and molybdenum requirement for growth of Methanobacterium thermoautotrophicum. Growth of Methanobacterium thermoautotrophicum on H2 and CO2 as sole energy and carbon sources was found to be dependent on Ni, Co, and Mo. At low concentrations of Ni (less than 100 nM), Co (less than 10 nM) and Mo (less than 10 nM) the amount of cells formed was roughly proportional to the amount of transition metal added to the medium; for the formation of 1 g cells (dry weight) approximately 150 nmol NiCl2, 20 nmol CoCl2 and 20 nmol Na2MoO4 were required. A dependence of growth on Cu, Mn, Zn, Ca, Al, and B could not be demonstrated. Conditions are described under which the bacterium grew exponentially with a doubling time of 1.8 h up to a cell density of 2 g cells (dry weight)/l.


Changes in sporulation potential during the growth cycle of Bacillus subtilis. The sporulation potential of Bacillus subtilis as a function of position in the cell cycle was determined by transferring cells from growth medium to sporulation medium at various times during growth. Growth was induced by incubating heat-activated spores in rich medium or by diluting stationary phase vegetative cultures with fresh growth medium. The results supported earlier observations that sporulation potential is cell cycle dependent. The rise in sporulation potential was studied by exposing cultures to the inhibitors of cell wall and protein synthesis, vancomycin and chloramphenicol. The delay in the appearance of the peak of sporulation potential caused by these inhibitors compared with the reported lack of effect of nalidixic acid, indicates that the appearance of sporulation potential requires synthesis of a macromolecular component other than deoxyribonucleic acid. The effect of nalidixic acid in preventing the decline of the sporulation potential was compared with the effect of high temperature on a mutant temperature sensitive for the initiation of DNA replication. It was found that prevention of chromosome completion with nalidixic acid maintained a high sporulation potential, whereas prevention of chromosome re-initiation in the temperature sensitive mutant did not affect the decline in sporulation potential as the cells enter stationary phase.


[A clinical study of the non-Hodgkin's lymphomas according to the Kiel classification (author's transl)]. A study of 138 patients with Non-Hodgkin's lymphomas was performed. The results were compared with retro- and prospective investigations of patients of the Kiel lymphoma study group. The aim of the study is to evaluate the value of the Kiel classification for the therapy and prognosis of malignant lymphomas. The malignant lymphomas of low- and high-grade malignancy differ significantly in their biological behaviour. Especially the clinical progress of the disease and the response to therapy are different. The lymphomas of low-grade malignancy may not need any treatment for long periods in some patients. In contrary, patients with lymphomas of high-grade malignancy need an active treatment immediately after the establishment of the diagnosis. In stages I and II radiotherapy is sufficient, in stages III and IV chemotherapeutic regimens are necessary. Each type of malignant lymphoma may be accompanied by leukemia. Most of the malignant lymphomas, particularly the immunocytomas, may produce monoclonal gammopathies, most of them being macroglobulinemias. Thus the macroglobulinemia Waldenström is just a clinical necessary to evaluate the significance of the subtypes of the Kiel classification for the planning of specific treatment.


[Cerebral seizures, alcoholism, and deliria (author's transl)]. The occurrence of cerebral seizures in alcoholics was investigated in case histories of 84 delirious and nondelirious male patients. Eighteen patients had seizures before they became alcoholics; the frequency of the seizures increased during abuse. Twelve had no deliria at all up to the moment of this investigation; in 66 of the patients the occurrence of seizures was assumed to be caused by alcohol abuse alone. Seventy-one patients had seizures irrespective of deliria and nearly 40% of them had no deliria at all. In 21% we observed only deliria with seizures; in 16% only deliria without seizures; and 24% had deliria both with and without seizures. The remaining 13 patients of 84 had only deliria complicated by cerebral seizures; only 3 had deliria without seizures. The seizures occurred as grand mal in 94% of the alcoholics, in all patients with genuine epilepsy, and in 60% of the patients with post-traumatic epilepsy.


Epidemiology of diarrhoeal diseases in Singapore. Diarrhoeal diseases in the form of bacterial food poisoning are prevalent in Singapore. Common food poisoning organisms implicated were Staphylococcus aureus, non-thypoid Salmonellae and Vibrio parahaemolyticus. El tor cholera, believed to be introduced through intra-regional trade and travel, occurred sporadically with a common source outbreak, probably food-borne, in September 1978. Shigellosis and amoebiasis were mainly confined to areas where poor personal hygiene was prevalent. The incidence of enteric favers (typhoid and paratyphoid) continued to decline with 39.1% of the cases imported in 1978. Non-bacterial diarrhoeal diseases, including rotavirus, constituted 8--90% of the cases. The emergence of multiple antibiotic resistance enterobacteria which gave rise to occasional outbreaks in institutions, is a matter of concern. Control of diarrhoel diseases is directed mainly at food control and legislation and health education, backed by a high standard of environmental sanitation and a well-established system of epidemiological survelliance. The use of oral rehydration solution for the treatment of diarrhoea is being promoted.


Experimental congenital infection of Toxoplasma gondii in Macaca arctoides. Congenital toxoplasmosis was studied in Macaca arctoides as a model for primates. Time-mated female monkeys were assigned to 5 experimental groups and inoculated with 4 different strains of Toxoplasma gondii before pregnancy and during various stages of gestation. All monkeys showed a rise in indirect hemagglutination antibody (IHA) titers following infection, and some had demonstrable parasitemia. Of the 23 progeny, 2 were stillbirths and 2 died soon after birth, but only the one neonate born of a mother inoculated intrauterally succumbed to acute toxoplasmosis. No physical anomalies were observed in any of the progeny. The toxoplasms organism was not isolated from any of the placentas obtained nor from the cord blood. None of the tissues from the stillborn or infant monkeys that died were positive for toxoplasmosis by mouse inoculation. The live progeny were usually born with high IHA antibody titers which were usually soon lost, indicating presence of passively acquired maternal antibody. However, 2 babies, born of mothers infected with tissue cysts on day 79 and 149 of gestation, maintained moderate to high titers for about 8 and 19 months respectively, indicating that an active infection had taken place but no clinical disease was detected. Data obtained suggest that although certain developmental stages of the toxoplasma organism and of the fetus may favor the occurrence of congenital infection, very little neonatal disease results in this primate model.


Serum complement (C3 and C4) levels in the Malaysian adult population. Serum C3 and C4 values were determined in 236 normal adults of three racial groups, using the single radial immunodiffusion techniques. The C3 levels varied from 47 to 119 mg/dl and C4 levels from 16 to 66 ml/dl (mean +/- 2SD). The values were found to be comparable to the normals reported in some Western series. No significant differences in the levels related to sex and race were found.


Pituitary response of prepuberal lambs to oestradiol-17 beta. In two experiments 48 prepuberal Merino ewe lambs were injected with oestradiol-17 beta (E2) or saline to study the effect of E2 on their plasma LH levels and on oestrus and ovulation. In the three groups which received 30 (experiment I), 50 and 30 (experiment II) microgram E2 respectively, 27 out of 28 lambs showed an LH response, the corresponding mean LH peaks being 64.3 +/0 22.5, 153.6 +/-33.4 and 91.7 +/- 16.9 ng/ml at mean intervals of 11.1, 11.2 and 10.5 h, respectively, after injection. None of the 20 lambs in the control groups had an LH level higher than 18 ng/ml 12 h after injection. In the three E2 groups, 41.7, 62.5 and 37.5% of animals showed oestrus within 26 h of injection while in the control groups only one animal showed oestrus. Of 13 animals showing oestrus in the E2 groups, 11 failed to ovulate. The mean pre-injection plasma FSH level in experiment I was 102.7 ng/ml, and in four 5--7-month-old lambs over several weeks uas 155.3 ng/ml. Despite these high pre-injection levels of FSH, it appears that the follicles were unable to respond to the LH peak which followed the E2 injection.


Isolation of inner cell masses from mouse blastocysts by immunosurgery or exposure to the calcium ionophore A23187. Two techniques have been evaluated for their use in routinely isolating inner cell masses from mouse blastocysts by destroying the trophectoderm. The most efficient method of immunosurgery was a 15-min incubation in a 1:50 dilution of rabbit anti-mouse spleen antiserum followed by a 30--60-min incubation in guinea pig complement (1:10). Alternatively, inner cell masses were isolated by incubating blastocysts in 10(-5) M calcium ionophore A23187 in medium devoid of calcium and magnesium ions. Inner cell masses re-exposed to immunosurgery or the ionophore were less susceptible to lysis than the trophectoderm had been. The presence of the zona pellucida reduced trophectoderm lysis by immunosurgery in antiserum dilutions greater than 1:100, but had no effect when in the presence of ionophore. Inner cell masses were consistently isolated from expanded blastocysts which had been collected 78 h after ovulation and cultured in vitro for 24 h before exposure to ionophore or immunosurgery, whereas blastocysts which had developed for the full 102 h in vivo were frequently unaffected.


Use of EDTA blood samples for mannosidosis testing. Preincubation of EDTA plasma with 4mM ZnCl2 restores acidic alpha mannosidase activity, previously inhibited as a result of chelation of Zn++ by EDTA. This allows estimation of acidic alpha mannosidase activity in EDTA plasma samples, followed by granulocyte tests on those samples with low activities.


Hexobarbital blood levels and effects on EEG in the presence and absence of caffeine. The interaction of caffeine and hexobarbital in the rat with spinal cord transection was studied. Duration of hexobarbital effect on the brain was taken as the time from the injection of hexobarbital (i.v.) to the return of pre-injection cortical voltage. Hexobarbital distribution and elimination was estimated by application of a two-compartmental model to values for blood hexobarbital concentration (determined by a direct gas chromatographic method after extraction). Caffeine caused a shift in the dose-response curve for hexobarbital but no changes in hexobarbital distribution and elimination. Results are interpreted on the basis of a central interaction of caffeine with hexobarbital at a brain receptor level.


[Antibacterial activity of mezlocillin and azlocillin/A survey (author's transl)]. A review of the antibacterial activity of 6-[(R)-2-[3-methylsulfonyl-2-oxo-imidazolidine-carboxamido]-2-phenyl-acetamido]-penicillanic acid sodium salt (mezlocillin, Baypen) and 6-[(R)-2-(2-oxo-imidazolidine-1-carboxamido)-2-phenyl-acetamido]-penicillanic acid sodium salt (azlocillin, Securopen) is given on the basis of extensive in vitro studies. The action of each of the preparations is compared with that of important antibiotics of other substance classes from the group of penicillins and cephalosporins.


[Ultrastructural and microbiological studies on the activity of azlocillin against Pseudomonas aeruginosa (author's transl)]. To examine the effect of 6-[(R)-2-(2-oxo-imidazolidine-1-carboxamido)-2-phenyl-acetamido]-penicillanic acid sodium salt (azlocillin, Securopen) on the ultrastructure of Pseudomonas aeruginosa investigations were carried out by electron microscopy on thin sections and on negative-stained preparations. Depending on the concentration and the contact time of azlocillin the bacteria showed distinct alterations. The bacterial cells did not build septa and therefore only grew in length up to 100-fold that of untreated controls. The bacteria diameter remained unchanged. Survival curves showed that these bacterial filaments were unable to build colonies. They were irreversibly damaged.


Pseudomonas infections of the lung treated with azlocillin. Microbiology, pharmacokinetics, clinic. 6-[(R)-2-(2-Oxo-imidazolidine-1-carboxamido)-2-phenyl-acetamido]-penicillanic acid sodium salt (azlocillin, Securopen), has a special activity against Pseudomonas species besides is broadspectrum character. We confirmed the high in vitro potency of that antibiotic in 200 clinical Pseudomonas isolates. Good penetration of azlocillin into bronchial secretions was found, especially through the inflamed bronchial mucosa, but also in the post-inflammatory phase. In 16 patients with serious Pseudomonas infections of the respiratory tract the good efficacy of azlocillin was shown. There were no tolerance problems.


The free amino acid pool composition during growth of the culture of Xanthomonas citri (Hasse) Dowson. Fourteen amino acids were found to constitute the intracellular amino compound pool of the developing culture of Xanthomonas citri (Hasse) Dowson. These were serine, dl-alanine, beta-alanine, leucine/isoleucine, threonine, aspartic acid, glutamic acid, asparagine, glutamine, cystine, histidine, tyrosine, tryptophan, and proline. Of these, beta-alanine, threonine, aspartic acid, glutamic acid, asparagine, and tyrosine could be traced as excretory amino acids in the bacterial culture filtrate, being exuded at the cost of and parallel to the depletion of their amounts in bacterial cell extract. Thus, the composition and pool size of intra- and extracellular amino acids varies considerably during the growth of this pathogen. The wall constituents of X. citri, especially alanine, glutamic acid, and aspartic acid, were prominent among the amino compounds detected.


The ultrastructure of Penicillium chrysogenum in the course of benzyl-penicillin biosynthesis. The find structures of high- and low-yield mutants of Penicillium chrysogenum, producing 100 and 10,000 units/ml of penicillin G, were compared. The cells of both mutants demonstrated a typical eukaryotic ultrastructure. In the cytoplasm nuclei, mitochondira, lipid bodies, endoplasmic reticulum, and Golgi vesicles were observed. In the cells of high-yield mutant, during the biosynthesis of penicillin, the number of lipid bodies decreased. It is possible that the lipids are metabolized in the process of biosynthesis of penicillin. In the cytoplasm more multivesicular bodies and small vesicles, about 40 nm in diameter, could be seen. These Golgi vesicles, present in largest number in cells of high-yield mutant, fuse with the cell membrane and play an important role in the transport of penicillin from the cytoplasm to the cell environment. The cell walls of the high-yield mutant become three times thicker during the antibiotic biosynthesis. No comparable changes were observed in the ultrastructure of the low-yield mutant. The cell wall thickness did not increase, the cytoplasm contained few Golgi vesicles only, and the lipid bodies can be seen in all cells.


The site of benzyl penicillin accumulation in Penicillium chrysogenum. Benzyl penicillin was localized in cells of Penicillium chrysogenum by means of enzymatical and immunological methods, enabling the determination of minute amounts of penicillin. The reactions were performed on ultrathin sections. They allow to determine the antibiotic inside of the cells. The results indicate that benzyl penicillin is present in the vesicles belonging to the Golgi apparatus. Benzyl penicillin is transported from the cytoplasm outside the cell membrane by the Golgi vesicles.


Effect of phenytoin on protein binding of valproic acid. In vitro experiments using the equilibrium dialysis technique were performed to determine the binding of valproic acid to plasma components in the absence and presence of therapeutic concentrations of phenytoin. The free fraction of valproic acid was found to be dependent on the total valproic acid concentration. Phenytoin did not influence valproic acid protein binding.


Sodium valproate in the treatment of cerebellar disorders. Because of the high concentrations of gamma-aminobutyric acid (GABA) in the cerebellar cortex and nuclei, an attempt was made to enhance GABAergic transmission in patients with cerebellar disease. Maximum tolerated doses of sodium valproate, a drug which inhibits the degradation of GABA, failed to influence cerebellar deficits in a double blind crossover study on six patients.


Fractionation of Plasmodium knowlesi-induced antigens of rhesus monkey erythrocyte membranes. In order to characterize parasite-induced host cell membrane antigens, the plasma membranes of Plasmodium knowlesi-infected rhesus erythrocytes have been compared with those of normal red cells and purified schizonts by immunochemical and biochemical techniques. Host cell membranes and schizonts were separated by differential centrifugation following nitrogen decompression. Isolated schizonts were further fractionated into several subcellular compartments. Crossed-immune electrophoresis, against monkey anti-schizont serum, of Triton X-100-solubilized material identified 7 P. knowlesi-specific antigens, of which 4 could be detected only in the host cell membranes. These membranes also contained 3 proteins, with relative molecular masses of 55 000, 65 000 and 90 000 and isoelectric points at pH 4.5, 4.5 and 5.2, respectively, which are lacking in normal membranes. Pulse-chase experiments with ((14)C)-glucosamine showed that these parasite-induced host cell membrane components are glycoproteins.


Immunization of rhesus monkeys with blood-stage antigens of Plasmodium knowlesi. Evaluation of the efficacy of 3 nonviable blood-stage antigens-schizont Ag, merozoite Ag, and "French press" Ag-of Plasmodium knowlesi, emulsified in Freund's complete adjuvant (FCA), was carried out in a study involving 32 rhesus monkeys. After 2 immunizations, administered 6 weeks apart, monkeys were challenged with a variant of P. knowlesi different from that used for immunization. All 8 control monkeys that received either PBS or FCA developed a severe parasitaemia and died of the infection within 12 days after parasite challenge. In the group that received the freeze-thawed schizont Ag, 5 of the 8 monkeys died at the same time as the controls, 1 died a day later, and 2 survived the infection (maximum parasitaemia: 0.7%; 3.2%). In the freeze-thawed merozoite Ag group, 1 monkey died at the same time as the controls, 5 monkeys died between 14 and 17 days after challenge, and 2 monkeys survived the infection (maximum parasitaemia: <0.01%; 2.7%). In the lyophilized French press Ag group, 2 monkeys died at the same time as the controls, 2 died between 12 and 15 days after challenge, and 4 survived the infection (maximum parasitaemia: 1.2%; 0.4%; 0.9%; 0.07%).Immunized monkeys failed to gain weight during the period of immunization and abscess formation at injection sites was comparable in monkeys of the 3 Ag groups. Reticulocytosis and anaemia developed in all monkeys that survived longer than 12 days after challenge, even in monkeys with very low-grade levels of parasitaemia.Various tests were performed to assess humoral or cell-mediated immune responses during the period of immunization and after challenge. No clear-cut relationship could be established between the results of any of these tests and the survival of immunized monkeys after parasite challenge. Some of the results, however, suggest that there might be some relationship between cell-mediated responses and the survival of immunized monkeys beyond the survival time of control monkeys.


Immunization of Aotus trivirgatus against Plasmodium falciparum with irradiated blood forms. Five owl monkeys, Aotus trivirgatus, were immunized against falciparum malaria by the intravenous injection of gamma-irradiated parasitized blood from donor monkeys with high parasitaemias. Each animal received approximately 1 x 10(8) parasitized erythrocytes per immunizing dose. Three monkeys received 4 doses and 2 monkeys received a single dose. After challenge, 3 control monkeys that received saline and the 2 monkeys that received a single dose of parasitized blood died with fulminant parasitaemias. The 3 monkeys that received 4 immunizing doses had either no parasitaemia at all (one monkey) or low-grade (<2.5%) parasitaemias (two monkeys); one of the latter monkeys died after the malaria had run a prolonged course. No striking haematological abnormalities were noted in animals as a result of the immunization procedure. These results confirm that immunity to Plasmodium falciparum can be induced in owl monkeys by inoculation with parasitized erythrocytes that have not undergone culture and without the use of adjuvant.


Duration of immunity following a single vaccination with irradiated sporozoites of Plasmodium berghei. A rodent model of sporozoite immunization against malaria based on a single immunizing dose is described. The duration of protective immunity was measured as a function of dose, and the results suggest that the percentage of protection follows a bimodal distribution. The first peak occurs between days 7-12 after immunization, while the second peak occurs at approximately 28 days. Although the percentage of protection declines steadily after the second peak, some immunity was detectable as long as 140 days after immunization with the higher doses.


Preliminary studies on vaccination of rhesus monkeys with irradiated sporozoites of Plasmodium knowlesi and characterization of surface antigens of these parasites. Studies were conducted to develop an effective method of inducing protection against sporozoite-induced malaria in a primate system and to obtain information regarding the surface membrane antigens of sporozoites. Immunization of rhesus monkeys was performed with gamma-irradiated sporozoites of Plasmodium knowlesi. Levels of antisporozoite antibodies were monitored by immunofluorescence, sporozoite neutralization, and the circumsporozoite precipitate reaction, and appeared to correlate well with protection. Only the intravenous route was effective in inducing both protection and antisporozoite antibodies. Immunization with sporozoites mixed with Freund's complete adjuvant failed completely to induce protection and resulted in a minimal antibody response. Mechanisms of resistance to sporozoites probably involve the interaction of the host's immune system with the parasite's surface antigen(s). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of surface-labelled, partially purified sporozoites followed by autoradiography revealed the presence of a small number of labelled proteins in the extract. Immunoprecipitation with specific antisera to P. berghei detected primarily one of these membrane components, with an apparent molecular weight of 41 000. The molecular weight of this main surface antigen in sporozoites of P. berghei was different from that in sporozoites of P. knowlesi.


Factors affecting the in vitro culture of Plasmodium falciparum and Plasmodium knowlesi. Plasmodium falciparum and Plasmodium knowlesi have been established in continuous culture using the basic method of Trager & Jensen. Various parameters of the culture system have been examined, namely, the gas requirements, serum and red cell requirements, frequency of medium replacement, and a comparison of static and agitated cultures made. The most important factors affecting growth in vitro seem to be the oxygen tension, red cell concentration, the frequency with which old medium is replaced, and the use of appropriate sera. Preliminary results indicate that horse serum may be possible as a replacement for human serum. Initial studies with P. knowlesi indicate that in the course of adapting to culture, parasites may change their antigenic specificity.


Immunization of experimental monkeys against Plasmodium falciparum: use of synthetic adjuvants. The replacement of Freund's adjuvant by a possible safe adjuvant for effective immunization of owl monkeys (Aotus trivirgatus griseimembra) against a human malaria parasite, Plasmodium falciparum, has been investigated. Experiments involved the use of two synthetic adjuvants: MDP (N-acetylmuramyl-L-alanyl-D-isoglutamine) and stearoyl-MDP (6-O-stearoyl-N-acetylmuramyl-L-alanyl-D-isoglutamine). In both cases, P. falciparum merozoites obtained through short-term in vitro cultivation were used as antigen. MPD was used as adjuvant in 5 owl monkeys; 2 control monkeys died and of the 3 experimental monkeys only 1 survived. In contrast, in another experiment where stearoyl-MDP was used as adjuvant, there was 100% protection of 4 immunized monkeys against a challenge with the homologous strain of P. falciparum. The results of the second experiment are encouraging for the development of an effective and safe vaccine for human malaria.


Malaria vaccination with irradiated sporozoites: serological evaluation of the antigen and antibody responses. Vaccination against Plasmodium falciparum with attenuated sporozoites is the goal of the US Navy's Malaria Vaccine Program. One requirement in the development of this vaccine is an immunological test to study the sporozoite antigen and immune responses it induces. Using an indirect fluorescent antibody test (IFAT) and P. berghei in the mouse or rat as a model, we have made significant progress toward this goal. Four antigens were detected in vaccine preparations: sporozoite-specific antigens, mosquito antigens, antigens on the sporozoite that are common to erythrocytic stages, and bovine serum albumm, an antigenic element of the isolation medium no longer employed. The IFAT was a reliable monitor of vaccination in a mouse and rat model in conjunction with protection to challenge. The test was a sensitive monitor of vaccine quality. Anamnestic responses to bites of infected mosquitos were detected in mice previously immunized with irradiated sporozoites.


Characterization of sporozoite surface antigens by indirect immunofluorescence: detection of stage- and species-specific antimalarial antibodies. Indirect immunofluorescence (IF) was used to localize stage-specific antigen(s) on the surface of the sporozoite membrane. The authors examined the feasibility of using an IF assay to determine whether an antisporozoite response is developed by individuals living in endemic areas. The specificity and sensitivity of the IF assay were first defined by using hyperimmune sera of sporozoite-immunized hosts protected against rodent (P. berghei), simian (P. knowlesi), and human (P. falciparum, P. vivax) malarias. Species- and stage-specific antisporozoite antibodies were detected by this technique when viable or glutaralde-hyde-fixed sporozoites were used as antigen. The sensitivity of indirect immunofluorescence was found to be 5-10 times greater than that of the circumsporozoite precipitation (CSP) assay. Preliminary studies on sera obtained from individuals living in endemic areas showed that the IF assay can be used to detect species- and stage-specific antisporozoite antibodies in sera of naturally-infected human and simian hosts.


In vitro inhibition of the growth of Plasmodium falciparum by Aotus serum. An in vitro assay for measuring growth and reinvasion inhibition of Plasmodium falciparum was developed from a microculture system. Inhibition of growth was observed after parasites had been incubated with Aotus serum obtained from monkeys that had become immune to malaria after repeated exposure to P. falciparum. Immune Aotus serum (IAS), at concentrations as low as 1.25%, showed a marked inhibitory effect upon parasites cultured in 10% normal human serum (NHS). Growth was also inhibited when 5% of normal Aotus serum (NAS) was added to medium containing 10% of NHS. The inhibitory activity of NAS, but not of IAS, could be removed by lowering serum concentrations below 2.5% or by exposing the serum to 56 degrees C for 30 minutes. Further investigations with relatively synchronous microcultures showed that (3)H-hypoxanthine, a nucleic acid precursor, was incorporated preferentially into the more mature parasites of P. falciparum. Additional studies with the microculture system should elucidate the effects of immune serum on different asexual erythrocyte stages and, in general, facilitate the study of immune effector mechanisms in falciparum malaria.


A study of cellular and humoral immune responses in owl monkeys (Aotus trivirgatus) following vaccination against Plasmodium falciparum. VACCINATION OF ANIMALS AGAINST MALARIA PARASITES IS THOUGHT TO INDUCE TWO BASIC IMMUNOLOGICAL RESPONSES: (i) specific recognition of parasite antigens by the host, and (ii) a generalized immune enhancement due to the presence of adjuvant. Immunological techniques were used in this study to monitor cellular and humoral immune changes in owl monkeys prior to and following immunization with a vaccine consisting of merozoite-enriched schizonts of Plasmodium falciparum and one of three adjuvants: N-acetylmuramyl-L-alanyl-D-isoglutamine (muramyl dipeptide), Freund's complete adjuvant, or 6-O-stearoyl-N-acetylmuramyl-L-alanyl-D-isoglutamine. The results showed that most of the immunized animals responded specifically to malarial antigens as demonstrated by the fact that peripheral blood lymphocytes underwent blast transformation in vitro in the presence of parasite antigens and that substantial antibody titres were produced as detected by indirect immunofluorescence. By use of the in vitro inhibition test, it was found that sera from immunized owl monkeys collected after challenge greatly inhibited merozoite reinvasion. Generalized nonspecific immune responses observed in owl monkeys following vaccination included an increase in the number of white blood cells and the proportion of T and B cells in the peripheral blood. Responses to mitogen stimulation with phytohaemagglutinin, concanavalin A, and pokeweed mitogen, however, did not appear to be appreciably affected by immunization.


Use of attenuated sporozoites in the immunization of human volunteers against falciparum malaria. Three human volunteers were successfully protected against sporozoite challenge by immunization with attenuated sporozoites of the Tamenie strain of Plasmodium falciparum from Ethiopia. The immunizing sporozoites were attenuated by exposing infected Anopheles stephensi mosquitos to X-rays at a dose of at least 120 Gy (12 000 rad). These irradiated, infected mosquitos were allowed to feed upon volunteers, thereby inoculating sporozoites into their blood stream. During the 10- to 38-week period of immunization, volunteers were exposed 6-8 times, at irregular intervals, to a total of 440-987 irradiated infected mosquitos. Protection against challenge by nonirradiated infected mosquitos lasted for at least 8 weeks, but not 16 weeks, after the last immunization with irradiated sporozoites. By contrast, volunteers who were exposed to a total of 200 or fewer irradiated infected mosquitos on 2-4 occasions were not protected upon challenge. Immunization by a sufficient number of irradiated mosquitos infected with the chloroquine-sensitive Tamenie strain from Ethiopia also protected against challenge with the chloroquine-resistant Marks strain of P. falciparum from Viet Nam. The results obtained in these studies suggest that immunization with attenuated sporozoites may be a useful method of protecting small groups of nonimmune individuals living in endemic areas. These findings should encourage further efforts to develop a sporozoite vaccine against human malaria.


Isolation of stages of the human parasite Plasmodium falciparum from culture and from animal blood. Procedures for isolation of various forms of the asexual erythrocytic stages of the human parasite Plasmodium falciparum are outlined. The procedures employ the plasma expander Physiogel, which is composed of a chemically modified, partially hydrolysed gelatin dissolved in Ringer's lactate. Based on the observation that parasitized cells which are easily separable by this technique differ appreciably at the ultrastructural level, a mechanism by which separation occurs is proposed.


The use of membrane screen filters in the isolation of Plasmodium berghei sporozoites from mosquitos. An improved procedure is presented for the isolation of Plasmodium berghei sporozoites from host mosquitos. The method employs filtration through a series of Nuclepore membranes followed by two consecutive centrifugations of the filtrate layered over Renografin-60 solutions of different densities. A Coulter Counter was used to compare isolations prepared by this technique with those prepared by a routinely employed discontinuous gradient method. When the sporozoite concentration in each preparation was standardized at 300 sporozoites per ml, isolations prepared by the new technique were significantly cleaner than isolations prepared by the discontinuous gradient method, containing an average of 1706 total particles per ml compared with 46 107 total particles per ml. The latter procedure was more effective, however, in removing viable microorganisms. Sporozoites isolated by both techniques were similar in immunogenicity and virulence.


Low-temperature preservation of sporozoites of Plasmodium berghei. Large numbers of biologically active sporozoites are needed as a source of potential antigen in the development of a malaria vaccine and the most practical method of accumulating sufficient numbers of these forms would be to freeze and store them at low temperature. The purpose of this work was to determine the feasibility of preserving the infectivity of frozen and thawed sporozoites. The results indicate that sporozoites of Plasmodium berghei exhibit a typical response to freezing over a wide range of cooling rates; the distribution around the optimum was a normal one and both the magnitude and position of the peak of their infectivity depended upon the preservative used. The optimum cooling rate with preservatives of high relative molecular mass was between 20 degrees C and 60 degrees C per min, but varied with the preservative used. A new apparatus was designed and built to yield reproducible controlled cooling rates over the range studied. A comparison of various commonly used preservatives showed mouse serum alone to be effective, but the combination of serum and hydroxyethyl starch proved to be the best cryopreservative of those examined.


Localization and chemical characterization of Plasmodium knowlesi schizont antigens. The identification of malarial antigens that induce protective immunity could provide a rational basis for developing an effective antimalarial vaccine as well as specific serodiagnostic tests indicative of clinical immune status. Since protective immunity is probably induced by stage-dependent rather than stage-independent antigens, the antigenic composition of different stages of Plasmodium knowlesi has been compared, and a limited chemical characterization undertaken. This information should provide some insight into the types of preparative procedure appropriate for the purification of functionally important malarial antigens.


[Enzymes catalyzing the reduction of aldopentoses and the oxidation of pentitols in Mycobacteria]. M. phlei, grown on synthetic Sauton medium (with 6% glycerol as carbon source), had NADPH- and NADH-aldopentose reductase, as well as NAD-pentitol dehydrogenase activities; some of their properties are studied. These activities are not present in BCG grown on the same medium. All experiments of aldopentose-reductase induction in BCG on a D(+)xylose medium were negative.


[Increase in activity of natural killer cells (NK cells) by serum thymic factor]. The serum thymic factor (FTS) and one of its structural analogues have been administered to adult normal Mice for 2 to 12 weeks. This treatment induced a significant increase in spleen cell natural killer (NK) activity, evaluated by a cytotoxicity assay against YAC cells.


[Measles and temporary increase in masculine births: coincidence or causality?]. Data from three West-African populations shows significant increase of sex-ratio. In two of them a considerable excess of male births came from conceptions the year following an epidemic of measles. This is limited to the villages affected by this epidemic. These facts seem to be similar to those related to hepatitis and sex-ratio. They suggest similarities between measles virus and surface antigens of Y sperms. This hypothesis could be tested by immunological investigation.


[Detection and localization of a ribonuclease activity associated with vaccinia virus]. A ribonuclease associated with vaccinia virus can be detected when reduced concentrations of nucleotides are used for an in vitro RNA synthesis assay. The non-viral origin of this ribonuclease may be inferred from its external location and from its variable activity on different purified virus stocks. The detection of this ribonuclease activity on purified virus grown without foetal Calf serum may suggest that this enzyme is of cellular origin.


[Identification of a virus similar to hepatitis B virus in non-A non-B hepatitis]. Hepatitis B virus-like particles (including DANE particles) with DNA polymerase activity but negative for HBs Ag have been identified in NON-A, NON-B hepatitis sera positive for HC Ag. Although specifically associated with the particles, HC Ag is not a surface antigen of the hepatitis C virus identified here for the first time. The relationship of this agent with HBV seems obvious, and deserves further study.


[Importance of respiratory route in experimental salmonellosis in Balb/c mice]. Infection of Mice with S. typhimurium, was conducted by respiratory, oral and peritoneal routes. The results obtained showed that infection occurred more quickly by i. p. than by other routes. However i. n. inoculation is the more efficient concerning Bacteria concentration per Mousse to obtain the DL 100. A study of Salmonella growth in vivo was conducted in lungs, spleen, liver and blood. The results are discussed in relation to the calf respiratory Salmonellosis actually observed.


[Diagnosis of the presence of meconium in amniotic fluid by dispersion curves of the longitudinal relaxations of protons]. Dispersion curves of the longitudinal relaxation T1 of protons in healthy amniotic fluid in a meconium solution are distinct at low Larmor frequencies (V0 less than 100 kHz). We are thus able to distinguish these fluids by T1 measurements in this range.


[Seasonal variations in the sexual activity of the male genet (Genetta genetta L.)]. The Genet testis shows a spermatogenic activity throughout the year. The plasmatic androgen level presents two periods where the values are higher; one in spring the other in autumn. The spring testosterone rise coïncides with a mating period.


[Morphological differentiation of neuroblastoma by 1 methyl cyclohexane carboxylic acid (CCA) and some C1 derivatives]. We describe the induction of neuroblastoma morphological differentiation by 1 methyl cyclohexane carboxylic acid (CCA) and by some C1 derivatives of CCA. This induction proceeds in a medium which allows, in the absence of inducer, a normal growth of neuroblastoma cells and contains 7.5% fetal Calf serum. In order to establish a clear-dose response relationship and to compare the relative potencies of different drugs, the morphological changes were assessed by examining "long neurite-bearing cells".


[Interpretation of the gingival inflammatory infiltrate]. The investigation of tooth eruption in dermoid cysts of the ovary allows one to observe in the chorion at the level of the epithelial attachment, an anatomical component that seems to correspond to a local system of defense. Up until now this has been considered to be a secondary inflammatory response resulting from the bacterial or mechanical assault. This component is found to form when the tooth erupts and constitutes an important element in the physico-pathogical process and of the immune response of the dental organ.


[Phospholipid and fatty acid composition of lamellar bodies isolated from chicken lungs at different stages of development]. In order to elucidate whether the lamellar body content represents functional surfactant as soon as these bodies appear in lung epithelium during fetal development, lamellar bodies were isolated from lungs of Chicken at various developmental stages. Their content was analyzed and compared to extra-cellular functionnal surfactant. The results show that this content undergoes gradual changes in the phospholipid/protein ratio, as well as in the relative phosphatidylcholine amount. Thus, in the Chicken, nascent lamellar bodies do not represent the final form of extracellular surfactant.


[Oculomotor reflex evoked by vibrassae stimulation in the cat]. The oculomotor effects of vibrissae stimulation have been studied in the "encéphale isolé" non-anaesthetized Cat. Intracellular recording of lateral rectus motoneurones showed an excitatory projection via the ipsilateral infraorbital nerve to these motoneurones. The afferent fibers are type A alpha, recuited with a low threshold (0.2-0.5 mA) and convey information about vibrissae displacement.


[Vitamin D and reproduction in Wistar rats]. Females have received from weaning a semi-synthetic diet  (0,35% Ca; 0,32% P; 0,03% Mg), with or without vitamin D (+/- D). Fertility of these females was not changed. At weaning, the young from mothers--D had lower weight, calcemia and phosphatemia, but the ash content of the femur was the same as for young from mothers + D. Young rats from mothers +/- D were given the diet with or without vitamin D. 19 days after weaning, the diet of the mother (+/- D) appears to be the principal factor for growth, calcemia and the ash content of the femur; the diet of the young (+/- D) produced only slight differences between ash contents.


[Radioautographic identification of dopaminergic axons in the rat CNS. In vitro marking with tritiated dopamine or noradenaline]. A new radioautographic technique for the visualization of central dopaminergic fibers is described. This technique is based upon in vitro uptake of triatiated norepinephrine or tritiated dopamine in the presence of monoamine oxidase inhibitor and specific uptake inhibitor in noradrenergic fibers. After a short step in glutaraldehyde, the use of osmium vapours as a fixative, instead of osmic acid immersion, allows a satisfactory retention of triatiated amines within dopaminergic fibers.


[Effect of progesterone on male and female sex behavior induced by steroid hormones in adult ovariectomized ewes]. Ovariectomized adult Ewes daily treated with I/M injections of testosterone propionate (10 mg/day) or oestradiol benzoate (200 microgram/day) show male-like sexual behaviour and simultaneously a permanent female receptivity. Additional daily treatment with 100 mg of progresterone results in a rapid decrease in male sexual responses both when Ewes are treated with testosterone and with oestradiol. No effect is observed on the female receptivity. Cyproterone acetate (100 mg/day) given to Ewes in addition to daily injection of testosterone propionate (10 mg/day) has effects similar to those of progesterone. These results show that progesterone has antiandrogen and antioestrogen properties and that this hormone administered at high doses is able to inhibit male activity.


[Lewis blood group activity in Fasciola hepatica]. Lewis blood group specificities are detected in the liver Fluke: Fasciola hepatica. Such specificities had never been reported in nature, other than in the human body. The investigation of this phenomenon might contribute to a better understanding of the mechanisms involved in parasite-host relationship.


Providing saline reacting anti-D cell typing reagent. Since 1969, when anti-D specific IgG was made available to protect recently delivered D negative women from sensitisation by fetal D positive cells, the supply of saline acting anti-D, suitable for reagent use has diminished. The same immunized male blood donors that provide the anti-D for the production of the specific IgG used for prophylaxis may, when bled up to fourteen days after restimulation, provide a source of saline anti-D suitable for reagent use. The results of such a procedure in 12 donors are described.


Growth, plaque assay and immunofluorescent studies on Tataguine virus in cell culture. The growth characteristics of Tataguine virus were studied in Cercopithecus monkey kidney (Vero); rhesus monkey kidney (LLC-MK2), baby hamster kidney (BHK-21); porcine kidney (PK-15), mouse fibroblasts (L-929) and Aedes albopictus cell monolayers. The virus replicated without producing any cytopathology in Vero, BHK-21 and Aedes albopictus: but not in the other three cell culture systems. Two or three subsequent serial blind passages in those cultures supporting the growth of the virus did not produce any appreciable increase in virus titre. Immunofluorescent staining of inoculated Vero cells demonstrated the presence of Tataguine virus antigen in the cytoplasm of infected cells. Plaques 1--1.5 mm in diameter were produced only in Vero cell culture. In neutralization tests performed on Tataguine virus, immune mouse and hamster sera, higher antibody titres were obtained by plaque reduction than mouse protection tests.


Respiratory function of blood of the avians. Individual blood samples were collected from wing veins of eight adult pigeons, two adult chickens and four adult geese and were analyzed for oxygen dissociation curves by hemo-scan, an instrument measuring PO2 and O2 saturation simultaneously. The findings agree with the common belief that avian blood in general has a low affinity for oxygen. Blood pH, PCO2, concentration of 2,3-DPG, lactic acid, pyruvic acid, red blood cell, hemoglobin and hematocrit were also determined, the related parameters have been calculated. Normal values of those variables were reported and the possible effects of some of those parameters on oxygen affinity are postulated.


Some problems in the bioassay of haematological drugs. The principles of biological standardization are well understood and generally accepted. However, discrepancies in potency estimates due to inter-method and inter-laboratory variations in assay hinder general agreement on potency values for certain biological drugs used in treating disorders of haemostatis. While attempts are made to minimize these differences, the magnitude of existing inter-laboratory discrepancies should be acknowledged.


Long-term effects of segmental alveolar osteotomy. The long-term effects of segmental alveolar osteotomy are reviewed. Thirteen patients attended a follow-up appointment between 5 and 9 years after operation and investigations related to the teeth, their supporting structures and the osteotomy bone cuts. The teeth involved in the osteotomies which gave a vital response after 1 year with ethyl chloride all maintained this response in the long term and about half the number of the teeth not responding at 3 years had recovered this response after 5 years or more. The incidence of periodontal pocketing, non-vital teeth and tooth mobility was low. There was radiographic evidence of the osteotomy bone cuts in more than half of the cases in the long term, although clinically, union had occurred in all cases. The occlusion was stable in all except one class II div. 2 case, which had been treated by lower segmental osteotomy only, where relapse had occurred.


Preprosthetic vestibular sulcus extension by the operation of Edlan and Mejchar. A 2-year follow-up study-I. Nineteen patients suffering from denture retention problems and/or pain on function due to atrophy of the mandibular alveolar ridge were operated on by the method of EDLAN & MEJCHAR. All patients felt an improvement of denture retention. Three out of seven still had pain on function after the operation. An average of 80% of the surgically created sulcus extension remained after 2 years.


Profile changes of bone and soft tissues following vestibular sulcus extension by the operation of Edlan and Mejchar. A 2-year follow-up study-II. The progression of alveolar resorption and the alteration of the soft tissue profile of the chin were studied in 15 patients operated on by the vestibular sulcus extension technique of EDLAN & MEJCHAR. Lateral cephalograms and tracings exhibited an apparently accelerated bone resorption of the alveolar crest. Furthermore, a significant reduction of lower lip height and increase of mental soft tissue protrusion have been recorded.


Swelling and trismus after third molar removal. A comparison of two techniques. The linqual split technique is associated with less external swelling than a method of mandibular third molar removal confined to a buccal approach.


Subperiosteal transmucosal porous ceramic/titanium implants. Clinical experience from three cases. Porous ceramic (Al2O3) material was used in combination with titanium fixtures as subperiosteal transmucosal implants in three patients at the Department of Oral Surgery and Oral Medicine, Dental Faculty, University of Oslo. Ceramic pieces were furnished with titanium receptacles and matching threaded posts. The implantation approach was based upon two stages, the first for subperiosteal placement of the ceramic onlay segment containing the titanium receptacles, and the second procedure for perforation of the mucosa and insertion of the posts intended for prosthetic use. A healing period of 8 weeks was planned between the two procedures. In one of the patients, however, the implants were removed only a few weeks after placement because of inflammation and soft tissue breakdown over the ceramic material. Reoperation was performed, however, with the same negative results. The other two individuals retained the implants, with uneventful postoperative healing periods. Between 6 and 8 weeks after mucosal perforation and insertion of the titanium posts, however, subjective and objective symptoms of infection were registered in both patients. In addition, one of the cases exhibited dehiscence and exposure of the ceramic. The implants were removed in both individuals.


Penetration resistance of dental injection needles. A laboratory test of the sharpness and penetration resistance of different brands of dental needles was undertaken. Both properties are essential when pain during injection is discussed. Using a Strograph in combination with a piece of stretched chamois leather resembling the mucous membrane, graphs were obtained for each needle tested and evaluated with reference to maximum penetration resistance at the actual point of puncture and to the smoothness of the further insertion through the leather. The findings gave a relative characteristic of each product based on a trial of 20 needles selected at random from each batch of 100 needles.


Ameloblastic fibroma.

Ameloblastic fibroma. Four cases of ameloblastic fibroma appearing in the posterior maxilla and the cupsid area of the mandibula are presented. One of the lesions appeared in the wall of a cyst and another was found in connection with a cyst. The treatment is conservative but the tumor needs close follow up for possible recurrence.


Incomplete branchial arch syndromes, branchial cleft cyst and vascular hamartoma in a patient with multiple neurofibromatosis. A case report with simultaneous occurrence of neurofibromatosis, incomplete branchial arch syndromes, a branchial cleft cyst and a pseudocyst in connection with a vascular hamartoma anterior to the right ear of a 38-year-old woman is presented. A possible common pathogenesis of the vascular hamartoma and the incomplete branchial arch syndromes as well as that of the neurofibromatosis is suggested. The pseudocyst is interpreted as a branchial cleft cyst showing inflammatory changes due to a pharyngitis shortly before the preauricular tumor appeared.


Abnormal root development, probably due to erythema multiforme (Stevens-Johnson syndrome). A case report is presented of a patient in whom nearly all teeth of the permanent dentition still present had short roots, while some teeth had never been formed at all. It may be concluded from the typical differences in length between the roots of the various teeth that this condition must be due to complete cessation of the growth of the teeth at the age of 7 or 8 years. Since the patient suffered a fulminant attack of erythema multiforme (STEVENS-JOHNSON syndrome) at this age, and since no other possible explanation of the short roots has been found, it is concluded that this clinical condition may have been the reason for the short root anomaly found. Damage or even destruction of the epithelial root sheath during the disease may be assumed to be the direct cause of the failure of full root development.


Benign osteoblastoma of the mandible. A clinical-pathologic review and report of a case. Benign osteoblastoma is an uncommon, solitary, osteoid and bone-producing tumor which is characterized by prevalent active osteoblasts and rich vascularized delicate fibrous stroma, previously regarded as malignant. The term benign osteoblastoma was recently proposed by different authors to separate this lesion from other solitary benign bone tumors. It most often involves long bones and vertebral column and other bones, and also occurs in jaw bones. There is a close histopathologic similarity between benign osteoblastoma and osteoid osteoma; consequently, much debate about them exists. Benign osteoblastoma has a good prognosis and is best treated by curettage or conservative surgical excision. Recurrence is rare. Because of its rarity in jaw bones, one case of benign osteoblastoma of the mandible is reported.


Immunological features and corticosteroid therapy of midline granuloma. This paper presents treatment of midline granuloma with corticosteroids and immunological features of this disease. The cellular immunity state of this patient was examined by counting T cell subpopulation and lymphocyte blastformation index (BI) by phytohemagglutinin at various time intervals during the corticosteroid therapy. Consequently, the percentage of T cells and BI became greatly increased according to the subsiding of the primary lesion, whereas non-T cell production and its function were depressed. In this communication we discuss a possible autoantibody mechanism and impared lymphocyte transformation in midline granuloma.


Splitting the condyle--a method of removing the condyle in cases of fibrous ankylosis. A method used to remove a fibrous ankylosed condyle is discussed. To overcome the complications of the conventional condylectomy, the risk of opening the neurocranium and laceration of the internal maxillary artery, the condyle is split in a latero-medial direction. Four patients have been operated upon successfully.


Enzyme-linked immunosorbent assay for immunological diagnosis of human tularemia. The enzyme-linked immunosorbent assay (ELISA) was applied for immunological diagnosis of human tularemia, using lipopolysaccharide from Francisella tularensis as antigen. Sera collected from patients, healthy individuals, and vaccinated volunteers were investigated for antibodies against F. tularensis by ELISA and tube agglutination. In ELISA all sera were titrated with a polyspecific anti-immunoglobulin enzyme conjugate. A limited number of consecutive sera from individual patients were also investigated for immunoglobulin G (IgG) and IgM antibodies by means of immunoglobulin class-specific conjugates. On an average ELISA was more than 10-fold as sensitive as tube agglutination. Two weeks after onset of disease, sera from patients had significantly higher titers in ELISA than sera from healthy controls. High titers persisted after more than 2 years. Significant amounts of both IgG and IgM antibodies were present within 1 to 2 weeks after infection. The antibody activity increased during the first month, without any significant change of the relation between IgG and IgM titers. After 2.5 years the IgG/IgM titer ratio of sera from patients was significantly increased. Within 6 weeks after vaccination sera from about half of the vaccinees had significantly elevated titers in ELISA. Titers observed after vaccination were generally lower than those found after infection. An elevated ELISA titer can be of diagnostic importance by the end of the first week of illness. A significant increase of titer in consecutive serum samples indicates a diagnosis of tularemia. Determination of IgG and IgM antibodies may be of value in determing whether a positive titer of a single serum sample is of longstanding or recent origin.


Effect of osmotic stabilizers on radiometric detection of cell wall-damaged bacteria. The effect of osmotic stabilizers on the 14CO2-dependent radiometric detection of cell wall-damaged Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa was studied in BACTEC 14C-labeled blood culture medium. The organisms were previously exposed to cefamandole or carbenicillin at 63 to 80% of the minimum inhibitory concentrations. The addition of 10% sucrose, 2.2% glycerol, and 2.2% ethylene glycol to the medium failed to reduce the time required for detection and diminished the amounts of 14CO2 released by the growing cultures. Viable counts made after 4 to 7 h of incubation showed a decreased culture density in osmotically stabilized media as compared with saline or Ficoll controls. Sucrose and Ficoll had little or no inhibitory effect on 14CO2 evolution by P. aeruginosa. The osmotic stabilizers tested did not seem to improve the survival of the bacterial inoculum and failed to increase the sensitivity of the radiometric system of detection.


Indirect hemagglutination test for detection of antibody to Rickettsia rickettsii in sera from humans and common laboratory animals. Antibody production in humans and three species of laboratory animals infected with Rickettsia rickettsii was determined with the indirect hemagglutination test. Rabbits, guinea pigs, and mice were inoculated with R. rickettsii and bled at intervals. Antibody which agglutinated both fresh and glutaraldehyde-fixed sheep erythrocytes sensitized with antigen prepared either from purified rickettsiae or from infected yolk sacs was found in rabbit sera at all intervals tested (10 to 59 days postinfection). Antibody which agglutinated fresh but not glutaraldehyde-fixed erythrocytes sensitized with either of the above antigens was detected in guinea pig sera obtained 7, 14, and 28 days postinfection. Antibody was found in mice inoculated with 5.6 x 10(6) plaque-forming units of R. rickettsii but not in mice given 5.6 x 10(2) plaque-forming units. Peak indirect hemagglutination titers occurred in nonvaccinated human Rocky Mountain spotted fever patients about 3 weeks after onset of illness, and antibody was still detectable after 1 year. Both human immunoglobulin G and human immunoglobulin M antibodies agglutinated sensitized cells, but immunoglobulin M antibodies apparently were more efficient. The indirect hemagglutination test is useful for the titration of human, rabbit, guinea pig, and mouse antibodies when the appropriate erythrocytes are used.


Adaptation of enzyme-linked immunosorbent assay to the avian system. A microplate enzyme-linked immunosorbent assay was developed to detect chicken anti-rovirus antibodies. Studies of the parameters which affect the outcome of the assay with avian serum revealed two aspects for a successful assay. First, enzyme-antibody conjugates prepared by the periodate oxidation technique were found to have retained far more immunological activity than conjugates produced by a glutaraldehyde cross-linking. Second, the results indicated an unusually high affinity of chicken immunoglobulin for the microplate plastic which was mostly eliminated by a pretreatment technique with fixed fetal calf serum. The enzyme-linked immunosorbent assay compared favorably with the latex passive agglutination test, yielding a titration endpoint of 1:511,000, or approximately 1,300 times more sensitive than the latex passive agglutination assay. The assay proved not only to be sensitive to less than 1 ng of specific antibody, but also to have low to moderate variance and high reliability.


Evaluation of a killed Rocky Mountain spotted fever vaccine in cynomolgus monkeys. A nonhuman primate model of Rocky Mountain spotted fever infection was developed in cynomolgus monkeys (Macaca fascicularis) infected by the subcutaneous route or by aerosol. Clinical responses, hematology and serum chemistry values, and pathological findings were similar to those found in humans ill with Rocky Mountain spotted fever. The clinical model was then used to test the efficacy of a killed Rocky Mountain spotted fever vaccine grown in chicken embryo cells. Monkeys were immunized with varying dilutions of the vaccine with a two-dose schedule and then challenged at 2 months with virulent Rickettsia rickettsii by the subcutaneous route or by aerosol. The undiluted vaccine totally protected monkeys against both challenges, even at extremely high doses.


[The outcome in pregnancies resulting after the induction of ovulation 519 pregnancies (author's transl)]. Some geneticists have been against induction of ovulation because there is a greater incidence of chromosome abnormalities in spontaneous abortions that occur after induced pregnancies. We have found the number of early abortions (15.6  p. 100) and of malformed fetuses (4.8 p. 100) is not greater than average when we have studied a personal prospective series of 140 pregnancies and a series of 519 pregnancies collected following an enquiry made among some of our colleagues. The number of late abortions and of premature labours is much higher (3.4 p. 100 and 14.8 p. 100 respectively) and they correspond particularly with the higher incidence of multiple pregnancies (12.3 p. 100) which have been obtained especially after the use of HMG. This has a direct repercussion on perinatal mortality (4.3 p. 100).


Ciliate protozoa in the rumen of Brazilian water buffalo, Bubalus bubalis Linnaeus. Total numbers and distribution of genera, subgenera and species were determined for the ciliate protozoa in rumen contents of 4 Brazilian water buffalo Bubalus bubalis Linnaeus. The fauna of one animal, housed in close proximity to European and zebu-type cattle, differed considerably from that of the remaining animals, which were somewhat isolated on a large ranch. Several of the protozoan species observed in the semi-isolated animals were first described in rumen contents from humped Indian cattle, and their subsequent occurrence in other hosts and geographic locations has been limited or absent. In all, 49 different species of protozoa were found, 8 of which have not been previously described. Three of the new species belong to the genus Entodinium: E. ciculum sp. n., E. spinonucleatum sp. n. and E. triangulum sp. n.; 4 to Diplodinium (Ostracodinium): D. (O.) brazili sp. n., D. (O.) esalqum sp. n., D. (O.) nucleolobum sp. n., and D. (O.) tiete sp. n.; and one to Diplodinium (Eudiplodinium): D. (E.) bubalus sp. n.


Extrusion of DNA from the macronucleus of Tetrahymena pyriformis GL. Administration of the thymidine analog 5-bromodeoxyuridine to exponentially growing cultures of Tetrahymena pyriformis GL in chemically defined medium results in inhibition of cell multiplication by at least one generation before DNA synthesis stops. Cell multiplication can be restored in these cultures, if they are transferred to fresh growth medium, but although most of the cells in the culture contain close to a G2-amount of DNA, a full DNA replication round is a prerequisite for renewed cell multiplication. Large extrusion bodies are found at the first division after transfer to fresh growth medium. Autoradiographic analysis has revealed that the DNA in the extrusion body is a representative of the DNA in the macronucleus indicating a random distribution of DNA between daughter nuclei and extrusion body.


Synchronization of division in vitamin B12-starved Euglena gracilis. Vitamin B12 deficiency arrests cell division in Euglena gracilis. B12 starvation for short periods made it possible to induce synchronous growth by addition of the vitamin. Culture conditions were established to optimize replenishment synchrony. The DNA content of E. gracilis in steady state culture and vitamin B12 deficiency culture was measured by flow cytofluorometry and was consistent with colorimetric determinations. The cell volume and DNA distributions of E. gracilis in synchronous culture were analyzed and the sequential changes during the division cycle were computed. Synchronous culture permits more definitive studies of shifts in cell volume and DNA distributions, in which the biochemical events required for cell division are presumably synchronized.


Absolute DNA determinations by flow microfluorometry. The DNA content of Euglena gracilis and murine leukemic cell line L-1210 were varied by growth under abnormal conditions. DNA measurements by colormetry and by flow microfluorometry were compared and absolute calibration of the flow device was established.


Antigenic analysis by direct immunofluorescence of 114 isolates of Rickettsia tsutsugamushi recovered from febrile patients in rural Malaysia. One hundred and fourteen Rickettsia tsutsugamushi isolates, recovered from febrile patients in central Peninsular Malaysia, were antigenically analyzed by direct immunofluorescence using eight prototype strains. Twenty-nine antigenic types were detected. The TA763, TA716, Karp and TA686 strains were the most common and occurred singly or in combination with each other or other strains in 86% of the isolates.


Comparison of experimental Rickettsia tsutsugamushi infections in silvered leaf (Presbytis cristatus) and cynomolgus (Macaca fascicularis) monkeys. Both silvered leaf and cynomolgus monkeys were infected with the Gilliam, Karp and Kato strains of Rickettsia tsutsugamushi. The two species developed similar clinical syndromes, but the antibody responses were greater in cynomolgus monkeys. In both species of monkeys, the Gilliam strain induced more severe clinical manifestations. At 10 months post-infection, silvered leaf monkeys were immune to homologous intradermal (id) challenge. Cynomolgus monkeys, at 15 months post-infection, were relatively resistant to homologous intravenous challenge, but not to a homologous or heterologous id challenge.


Effect of vanadate on gill cilia: switching mechanism in ciliary beat. Lateral (L) cilia of freshwater mussel (Margaritana margaritifera and Elliptio complanatus) gills can be arrested in one of two unique positions. When treated with 12.5 mM CaCl2 and 10(-5) M A23187 they arrest in a "hands up" position, ie, pointing frontally. When treated with approximately 10 mM vanadate (V) they arrest in a "hands down" position, ie, pointing abfrontally. L-cilia treated with 12.5 mM CaCl2 and 1 mM NaN3 also arrest in a "hands down" position; substitution of 20 mM KCl and 1 mM NaN3 causes cilia to move rapidly and simultaneously to a "hands up" position. The observations suggest that there are two switching mechanisms for activation of active sliding in ciliary beat one at the end of the recovery stroke and the other at the end of the effective stroke; the first is inhibited by calcium and the second by vanadate or azide. This is consistent with a model of ciliary beating where microtubule doublet numbers 1, 2, 3, and 4 are active during the effective stroke while microtubule doublets numbers 6, 7, 8, and 9 are passive, and the converse occurs during the recovery stroke.


Glycosaminoglycans of the plasma membranes of renal tubule and liver cells. Glycosaminoglycans were isolated from plasma membranes of hepatic and renal tubule cells of guinea pig. Plasmalemma of renal tubule cells contained more total glycosaminoglycans, hyaluronic acid, chondroitin-4 sulfates and chondroitin-6 sulfates, and less dermatan sulfates and heparin sulfates than liver plasma membranes. These glycocalyx components, owing to their polyanionic properties, may have a role in the transport of water, ions, and macromolecules across the cell membrane.


7-Acetylcytochalasin B: differential effects on sugar transport and cell motility. Cytochalasin B (CB) is a potent inhibitor of sugar transport and cell motility in animal cells. We have synthesized and characterized the CB derivative 7-acetylcytochalasin B (CBAc) and have found that it has differential effects on transport and motile processes in fibroblasts. The derivative inhibited sugar transport in human red cells, 3T3 cells, and chicken embryo fibroblasts at micromolar concentrations, although it was less potent than its parent compound. Unlike CB, which causes fibroblasts to round up and arborize at less than 10 microM, CBAc had no effect on fibroblast morphology and membrane ruffling at concentrations as high as 90 microM. Competitive binding experiments using [3H] CB showed that the affinity of CBAc for sites related to sugar transport in the red cell membrane is about one-fourth of that of CB. In contrast, similar experiments using [3H] dihydrocytochalasin B (a derivative which inhibits cell motility but not sugar transport) showed that the affinity of CBAc for sites associated with red cell spectrin and actin is only about 1/20 of that of dihydrocytochalasin B. This study demonstrates that acetylation of the C-7 hydroxyl group of CB reduces its effect on cell morphology and motility much more than its ability to inhibit sugar transport. This observation, together with our earlier work with dihydrocytochalasin B, establishes that the pharmacologic effects of CB on fibroblasts result from the binding of the drug to two distinct classes of receptors and that these receptors interact with different parts of the cytochalasin molecule.


Computer-assisted mapping with the light microscope. With the system described specific features in a histological section such as neuronal degeneration, specific cell types, or the silver grains of autoradiography can be accurately localized. It is simple to operate, and allows for data storage and retrieval. The system consists of a Zeiss Universal microscope equipped with stepping motor drives on the X and Y axes and interfaced to a PDP-12 computer equipped with a printer/plotter. The user controls the system through a joystick control box and teletype. In spite of magnification changes all points in the field remain constant relative to a point of origin selected at the beginning of the mapping sequence. Therefore, the outline and major features of a section are entered at low magnification while the detail of neuronal degeneration, for example, is added in its precise location at a higher magnification. The plotting of points can be done in either random fashion or in a systemized series of horizontal or vertical scans. A graphic display is available throughout the procedure and all maps are both printed on a Versatec printer/plotter and stored on a disk for future recall and analysis. Use of the system in plotting neuronal degeneration is described.


A method of reversible inactivation of small regions of brain tissue. A method is described for reversibly inactivating small, precisely localized regions of brain tissue with injections of nanoliter quantities of the local anesthetic lidocaine hydrochloride. The injections are made through a combined recording-injection probe consisting of a glass micropipette onto whose outer surface is plated a metallic cylinder for recording extracellular action potentials. The recording cylinder for recording extracellular action potentials. The recording cylinder, located a known distance from the pipette tip, picks up a continuous, large-amplitude, multiunit response which can be used to accurately position the tip according to physiological criteria. It also provides a means of determining the duration of the anesthetic block and estimating its spreat. This devise has been used to selectively block 200--400 micrometers regions of individual laminae of the lateral geniculate nucleus centered within 50 micrometers of a retinotopically defined target site. The blocks last from 3 to 10 min and can be repeated many times at the same location.


Discrepancies in the extracellular space of sympathetic ganglia measured using different isotopes of mannitol and sucrose. The extracellular space of rat superior cervical ganglia in vitro was measured using mannitol and sucrose labelled with tritium and carbon-14. The volumes of distribution of the 3H-labelled derivatives, especially [3H]mannitol, exceeded those of the 14C-derivatives. The divergence increased with increasing lengths of incubation. Thus, after 30 min incubation, 'spaces' (ml . g-1) were: [14Cu]mannitol, 0.407; [3H]mannitol 0.447; [14C]mannitol, 0.458; [3H]mannitol, 0.645; [14C]sucrose, 0.430; [3H]sucrose, 0.497. Using thin layer chromatography, it was shown that an average of 22% of the label in ganglia incubated for 120 min with [3H]mannitol, but only 4% with [14C]mannitol, was not associated with the parent compound. Both [3H]- and [14C]sucrose appeared to be metabolized by 11%. It is concluded that mannitol and sucrose can be metabolized in isolated ganglia and that this may lead to substantial errors in estimating the extracellular space, particularly when [3H]markers are used.


A dual-purpose device for protecting brain implants of baboons. A device is described which is relatively easy to construct and which can be used to protect brain implants affixed to the skull of large primates. The device is essentially a stainless steel ring cemented around the brain implant. A plexiglass dome can then be attached to the ring to protect the implant. Further, a second device can be constructed which attaches to the ring and which provides gentle leverage to the brain implant to remove adhered obturators.


Influence of A-factor on the ultrastructure of the A-factor deficient mutant of Streptomyces griseus. The fine structure of the A-factor deficient mutant strain of Streptomyces griseus grown in the absence or in the presence of this factor was studied by transmission electron microscopy. The submerged 96 h culture grown in the presence of the A-factor was characterized by the formation of a large amount of spores accompanied by mycelium disintegration. A number of essential ultrastructural changes were observed in the mycelium in the course of development and streptomycin biosynthesis after the A-factor addition. Thus, the cell wall was transformed in another manner at various developmental stages. For a long time the associated ribosomes were preserved. The number of intracytoplasmic membrane structures increased. New, unusual organelles appeared which consisted of parallel cylindrical structures. The possible nature of these organelles is discussed.


Serological properties of cellular and extracellular glycerol teichoic acid antigens of Streptococcus mutans. Cell-free supernatant of Streptococcus mutans cultures was concentrated by 50% ammonium sulphate precipitation to 1/50 of the original culture volume. Most extracellular glucosyltransferase activity was detected in the supernatant. Furthermore, the supernatant reacted with serotype specific antiserum, showing immunological identities with the serotype specific polysaccharide antigen obtained from whole cells by hot saline or NaOH extraction in double diffusion tests. Immunoelectrophoretic analysis also revealed that the supernatant antigen possessed significant quantities of lipid-bound and free glycerol teichoic acid antigens. The lipoteichoic acid (LTA) was obtained from whole cells by phenol-water extraction. The lipid-teichoic acid linkage was easily split by mild acid or alkaline hydrolysis. The supernatant and phenol antigens spontaneously sensitized sheep red blood cells. These antigens were adsorbed to hydroxyapatite powder. The extracellular LTA and glucosyltransferase were found to be closely associated and difficult to separate chromatographically. In vivo significance of these properties of LTA and GTase is discussed in terms of the ecology of S. mutans and other oral streptococci.


Effect of ions on the activity of the enzyme alkaline phosphatase from Plectonema boryanum. Alkaline phosphatase extracted from P. boryanum with lysozyme or polymyxin B treatment was used in a comparative study of cell bound and cell free enzyme. The effects of various ions on enzyme activity were tested. Calcium was found to enhance activity to the greatest degree stimulating the cell bound alkaline phosphatase 100% and cell free enzyme four-fold. Magnesium and potassium also stimulated the activity of cell bound and cell free enzyme. Other ions were found to be inhibitory to varying degrees.


Effects of microwaves on organisms. Ptychodiscus brevis--Gomphosphaeria aponina interactions. The two referenced organisms have been subjected to microwave irradiation (2,450 MHz CW). Initial experiments indicated no statistically significant loss of cell numbers as a result of simple temperature rise (6 degrees C) or experimental manipulation. Cell survival for P. brevis one week after irradiation was related to total energy absorbed, not power level used. Over a 48 h period, a first-order decrease in cell numbers was observed. All cells were destroyed at a threshold level of 0.1 kJ/cm3. In contrast, the blue-green alga G. aponina in log phase of growth showed no loss of cells. In the presence of G. aponina, P. brevis suffered cell lysis, though G. aponina thrived. Irradiation of the mixed cultures appeared to enhance the lysis effect.


Salmonella choleraesuis proteins and their relation to proteins from bacteria of heterologous sero-groups. A diversity of proteins was identified in the material isolated from S. choleraesuis with the help of sera prepared in rabbits with this material. The sera displayed, in agar-gel diffusions, numerous superimposed precipitation lines against proteins from: Salmonellae, Shigellae and E. coli. In contrast to proteins from S. paratyphi C, sharing identical identical 'O' 'factors, the serological activity of the S. choleraesuis proteins was impaired by heating. The immunochemical analysis of the sera before and after exhaustive absorptions with heterologous proteins exhibited a stronger relation of S. choleraesuis with S. thyphimiurium and S. Newport than with S. paratyphi C. The antibodies induced against free proteins with S. paratyphi C specificity, present in the mosaic of proteins isolated from S. choleraesuis, were removed by the respective absorption without substantial modifications of the homologous precipitation. In contrast, the absorption of the serum with proteins from either S. newport or S. typhimurium removed almost all the homologous induced antibodies. The strong relations found among species belonging to different serogroups underline the non-conformity of the empirical established serofactors.


Interaction of the mannosephilic lectins of Pseudomonas aeruginosa with luminous species of marine enterobacteria. The marine bacteria Beneckea harveyi and Photobacterium leiognathi were shown to bear mannose-containing binding sites for the mannosephilic lectins of Pseudomonas aeruginosa and concanavalin A (Con A). The interaction between the lectins and the marine bacteria was demonstrated by the bacteriagglutination test, by adsorption of the lectins onto the bacteria and by mannose-specific peroxidase-binding to the lectin-coated bacteria. Treatment of the bacteria with formaldehyde, phenol, ethanol or boiling them for 15 min, did not alter their ability to adsorb the lectins. The growth rate of the marine bacteria was unaffected when either the Pseudomonas lectins or Con A was added to the culture medium.


Effect of varying concentrations of EDTA on the antimicrobial properties of cutting fluid preservatives. The potentiating effects of EDTA and its disodium, trisodium and tetrasodium salts were studied in combination with several cutting fluid preservatives. It was found that a ratio of 1 part preservative to 2 to 4 parts chelating agent produced marked increases in antimicrobial action.


Influence of potassium chloride upon the binding and antibacterial activity of chlorhesidine diacetate and (ethoxy)5 octyl phenol (Triton X45) towards Bacillus megaterium KM-. Chlorhexidine (0.5-0.65 microM) and Triton X45 (30-40 microM) added to exponential phase Bacillus megaterium KM- cultures was growth inhibitory. The presence of KCl (0.05-0.35 M) in the medium did not significantly affect growth rate in the absence of drug, yet reduced the growth inhibitory activity of the chlorhexidine and enhanced that of Triton X45. These effects were maximal at KCl concentrations of 0.2 M and above, when complete protection towards chlorhexidine and lysis of the cultures in the presence of Triton X45 were observed. Time-survivor curves in the presence of chlorhexidine (0.7-1.0 microM) gave LT90 values of 1.5-2.0 h in the absence of KCl, yet its inclusion (0.35 M) totally inhibited this low level bactericidal activity. Drug absorption by whole cell and isolated cell wall preparations was determined in the presence and absence of KCl (0.35 M). Chlorhexidine uptake by intact cells was reduced by approximately 50% in the presence of salt whereas that of Triton X45 increased by a similar fraction. Uptake of chlorhexidine by the cell wall fraction accounted for approximately 50% of that for the whole cells and was relatively unaffected by the presence of KCl. Conversely, absorption of Triton X45 by the cell wall fraction accounted for most of the uptake by whole cells and increased markedly in the presence of salts.


Variations in attachment of Neisseria gonorrhoeae to vaginal epithelial cells during the menstrual cycle and early pregnancy. An in vitro system was used to study the ability of virulent gonococci to adhere to vaginal epithelial cells obtained from healthy donors during the pre- and postmenstrual phases, and from those in early pregnancy. It was found that more gonococci adhered to the cells from donors in the postmenstrual phase than to cells from those in the premenstrual one. This difference was statistically highly significant. The attachment rate of gonococci to vaginal epithelial cells was similar in early pregnancy and in the premenstrual phase.


[Studies on tissue affinity of serogroups of Pseudomonas aeruginosa (author's transl)]. The distributions of serogroups of 316 Pseudomonas aeruginosa strains isolated from respiratory secretions (134), urines (105) and wound secretions (77) of patients in various hospitals show significant accumulations of 0-groups 1 (wound secretions), 4 and 5 (respiratory secretions). The statistical prevalence of these serogroups is due to their overproportional frequency in specimens from particular hospitals and, therefore, caused by local epidemiologic situations. There is no evidence that P. aeruginosa strains with certain 0 antigens have a special affinity to wounds, the respiratory, or the urinary tract.


Properties of [3H]taurine release from crude synaptosomal fractions of rat cerebral cortex. The release of previously accumulated [3H]taurine and [14C]GABA from crude synaptosomal (P2) fractions isolated from rat cerebral cortex was studied using a superfusion system. The spontaneous efflux of [3H]taurine and [14C]GABA was stimulated by elevated concentrations of K+ (15--133 mM) in a concentration-dependent manner. This K+-stimulated relase of [14C]Gaba but not of [3H]taurine was enhanced in the presence of Ca2+. However, addition of 3 mM Ca2+ to the superfusion medium in the presence of the ionophore A 23187 resulted in a stimulation of the release of both [3H]taurine and [14C]GABA. These results are discussed in connection with the cellular localization of taurine in the central nervous system.


Myosin autoantibodies reacting with selective muscle fiber types. Myosin in striated muscle exists in at least two immunologically distinct forms. Human autoantibodies specific for either form show selective muscle fiber reactivity. One group of antibodies reacts with type 2b (white) muscle fibers, the other with type 2a (red) and type 1 (intermediate) fibers. All of these antibodies react with the A band of glycerinated myofibrils. Both groups of antibodies react with cardiac muscle. Muscle-fiber-specific antibodies are detected in approximately 1% of sera submitted for diagnostic screening. Sera from eight patients were studied further. The two patients with antibodies against type 2b (white) muscle myosin had rheumatoid arthritis. The six patients with antibodies against type 2a (red) and type 1 (intermediate) myosin had clinical features which included various autoimmune and infectious disorders.


The occurrence of new retinoblastomas after treatment. New separate tumors have developed in 11% of eyes containing retinoblastoma treated primarily with external beam radiation. The tumors were primarily found in the inferior retinal periphery and averaged 1.5 disc diameters in size. All of the tumors were controlled, with photocoagulation and cryopexy being the most frequently used treatments.


Neuroanastomosis of orthotopically transplanted palmaris longus muscles. Palmaris longus (PML) muscles of rhesus monkeys were transplanted, with or without anastomosis of the median nerve, to the nerve stump of the autograft. Because PML autografts revascularize spontaneously, vascular anastomoses were not performed. Muscle fibers regenerated in all autografts with neuroanastomosis, but in only three of eight autografts without neuroanastomosis. Five autografts without neuroanastomosis were replaced by noncontractile connective tissue. Growth and differentiation of muscle fibers into three fiber types and development of capillarity were analyzed histochemically, and succinate oxidase activity of whole-muscle homogenates was determined. None of these measures reached values for control PML muscles within 100 days of transplantation. In comparison to control muscles, autografts had slower times to peak tension and less absolute tension, but similar tension per square centimeter of muscle fiber cross-sectional area. Monkey PML autografts with neuroanastomosis were similar in structure and function to cat extensor digitorum longus autografts that had not had neuroanastomosis.


[Demonstration of a serum and liver antigen in acute or chronic non A-non B viral hepatitis]. A new precipitating antigen-antibody system possibly was demonstrated by immuno-diffusion in the serum of patients suffering from non A-non B hepatitis. The antigen appears during the first four weeks of transaminases elevation. In acute cases was transient antigenemia (average 3 weeks). Antibodies appeared rapidly after the disappearance of antigen. The same antigen was also detected, by immunodiffusion and by immunofluorescence, in the liver nuclei of infected hepatocytes. This antigen specific appears for non A-non B hepatitis since it is neither found in the serum of normal subjects nor in that of patients with cirrhosis toxic or viral that hepatitis A or B. The hypothesis of a virus associated antigen is the most likely explanation.


[Economic aspects of varices surgery. A plea for ambulatory treatment]. Health expenses have increased in recent years in the majority of western countries. The treatment of varicose veins of the lower limbs occupies an important position from the budgetary point of view, and the duration of hospitalisation is clearly a determining factor. Apart from other methods currently being investigated--Dextran, intermittent pneumatic compression, anti-aggregants--low dose heparin therapy appears to be 80% effective in the prevention of post-operative thromboembolic complications. In the same way local complications after varicose surgery--haematomas, lymphoedema--have disappeared, as a result of progress in surgical technique (fine, atraumatic dissection, rigorous haemostasis) and improvement in methods of bandaging. In the light of our last fifty ligations and stripping of the internal saphenous vein, notably without thromboembolic complications, it seems reasonable to consider short hospital stays, of three days, with the exception of "high risk" patients. "Out-patient" treatment (24 hours) can be considered for patients who are "active and motivated".


[Results of personal experience in the long-term treatment of chronic lymphatic edema]. The author sets out his comments on the satisfactory results obtained in the treatment of lymphatic oedema of the lower limbs. He puts much stress on the need always to undertake these treatments, which even when instituted late, bring about a definite improvement. Long term surveillance is absolutely indispensable.


[Critical study of the therapeutic effects of phlebotropic drugs on chronic venous insufficiency]. The authors describe two types of test. Their aim was to verify the therapeutic potency of drugs used in the treatment of chronic venous disorders. The tests were carried out using both a single prescription and using successive prescriptions. The results showed that with strict clinical surveillance, phlebotropic drugs had definite therapeutic effects both on the subjective disturbances and on certain objective parameters.


Dentin formation in formocresol pulpotomized primary monkey teeth studied by tetracycline and 3H-proline incorporation. The influence of formocresol treatment on the pulp tissue of 24 primary monkey teeth was studied using tetracycline and 3H-proline labeling techniques. Six untreated monkey teeth served as controls. The tetracycline labeled teeth were examined between 352 and 600 d, following treatment. The 3H-proline labeled teeth were observed over a period of 22-607 d, the isotope being administered 15 d before extraction. The labeling was evaluated in the coronal, middle and apical area of the roots. Dentin formation as indicated by tetracycline labeling was observed in both control teeth and successfully treated teeth, as well as in some of the unsuccessfully treated teeth. The average dentin formation rate per day varied from 1 micrometer in the control teeth to 0.14 micrometer in the pulpotomized teeth (P less than 0.01). Success of treatment was of significant importance for the amount of dentin formation (P less than 0.001). Labeling with 3H-proline, indicating collagen synthesis, could be observed in the pulp and predentin of the majority of areas judged to be normal, degenerated or inflamed. Labeling was not observed in fixed or necrotic tissue. In degenerated pulp tissue the proline labeling was clearly reduced. The findings indicate that dentin formation and collagen synthesis may take place in formaldehyde influenced pulp tissue although at a decreased rate.


A scanning electron microscopic study of the effect of various cleansing agents on cavity walls in vitro. The appearance of prepared cavity walls after treatment with different cavity cleansers and cleaning procedures was studied. Class II and V cavities were prepared on extracted human and monkey teeth and the cavities were exposed to the different agents for 1 min. Where commercial products were used, the manufacturers' directions for use were followed. The cavity walls were examined in the scanning electron microscope and representative photos were taken from dentinal and enamel areas. Only demineralizing solutions producing an etch pattern were able to remove the smeared layer from the cavity walls.


Microleakage along composite restorations in human premolars after treatment with different cleansing agents. Cavities prepared on extracted human premolars wereeee treated with different cleansing agents and by different procedures. The cavities were filled with a composite restorative material and microleakage was assessed by recording the penetration of basic fuchsin dye. Only acid agents which removed the smeared layer, producing an etch pattern on the enamel cavity walls, were able to prevent microleakage.


The epidermal melanocyte system in patients with malignant melanoma. Quantitative and qualitative investigation of dopa positive melanocytes. The results from this study indicate that in 5 out of 22 patients increased activity of the melanocytes in the vicinity (a few mm to 1 cm) of the tumour is to be found. In 8 out of 22 patients suspect cells were also found a few mm to 1 cm from the tumour; a finding which accounts for the frequent recurrence following close excision of the melanoma. Furthermore, it was ascertained that the total melanocyte population is increased in 14 out of 22 patients. An increased melanocyte population can contain a greater number of instabile cell forms that can easier be malignantly transformed under the influence of external factors.


Prolactin secretion and nodular goiter. (Hypothetical correlations). In a group of 129 female patients with nodular goiter, Graves' disease and primary hypothyroidism, the PRL and TSH secretions were studied in parallel. Concurrent changes in the two hormones were evident only in conditions of marked hyper- or hypothyroidism. In the patients with non-toxic nodular (single or multiple) goiter the TSH showed low or normal values, while the PRL appeared normal or elevated. From these results two important conclusions can be drawn: 1. the T3 and T4 levels interact with PRL secretion concomitantly with TSH only when they undergo a huge deviation from the normal range; 2. the goitrogenic action of PRL that has been reported in experimental animals cannot be excluded in man.


Antibodies to Candida species after operations on the large intestine: observations on the association with oral and faecal yeast colonization. The occurrence of agglutinating and precipitating antibodies to Candida species was determined in 50 patients undergoing operations on the large intestine. The number of patients with oral or faecal yeast colonization and the amount of faecal colonization increased during the period after operation. No patient developed overt fungal infection but 15 patients developed precipitins to mannan antigens and 9 patients developed percipitins to cytoplasmic antigens during this period. The association between the formation of precipitins and the extent of faecal colonization is discussed.


Enrichment of human heterokaryons by Ficoll gradient for complementation analysis of iduronate sulfatase deficiency. Ficoll gradients have been used to enrich for heterokaryons in cultures of human skin fibroblasts following polyethylene glycol (PEG) induced fusion. These gradients provide a simple and consistent method for obtaining populations of multinucleated cells, at least twofold greater than those resulting from fusion alone. Formation of glucose-6-phosphate dehydrogenase (G6PD) heteropolymers has been used as a functional assay for the presence of heterokaryons. Analysis of cell populations enriched for multinucleated cells has revealed complementation leading to iduronate sulfatase activity in heterokaryons derived from iduronate sulfatase-deficient fibroblasts expressing the Hunter and multiple sulfatase-deficiency mutations.


A simplified, cost-effective approach to the diagnosis of pheochromocytoma. Plasma catecholamines have been used for diagnosis and computerized tomography scanning for localization in 21 patients with proved pheochromocytoma. Blood catecholamines were obtained in 20 of the patients and were elevated significantly in every instance. Successful localization of the tumor was accomplished by excretory urography in only 47% of the cases, by selective angiography in 85% and by abdominal computerized tomography scanning in 88%. In 7 patients preoperative identification and localization of the tumor were determined correctly by computerized tomography scanning alone. Data in our series suggest that the diagnosis and localization of pheochromocytoma can be accomplished usually by a simple blood test and body scan. More elaborate and sometimes hazardous studies should be necessary only in exceptional cases, usually with tumors less than 3 cm. in diameter.


An experimental approach to the study of intraocular Toxocara canis. An experimental study of nematode endophthalmitis due to T canis and review of the literature has been presented. Six owl monkeys were infected either by nasogastric tube using embryonated T canis eggs or by carotid or intravitreal injection of second stage larvae. The clinical manifestations, especially ocular, were observed and various diagnostic tests performed. Only minimal or no intraocular changes were seen after systemic infection but significant abnormalities such as retinal hemorrhages and venous dilation were noted after intravitreal infection. Motile larvae were observed in the lenses of three eyes and in the vitreous of five eyes and, probably a sixth, after intravitreal injection. The intensity and timing of the intraocular reaction seemed to correlate with the infecting dose and apparent disappearance of larvae from the eye. Pathologic confirmation of larvae in the lens was obtained in one eye. A marked inflammatory reaction occurred in eyes receiving intraocular infection but none was seen in eyes with only systemic infectin. Various laboratory and serologic studies were performed, including the ELISA test, which were used to evaluate systemic as well as intraocular responses to infection with T canis. The two monkeys infected by nasogastric tube gave a positive ELISA response in the serum but intraocular fluids gave a negative response in all monkeys including those infected syst:mically and/or intraocularly. Problems in the understanding of clinical aspects of the disease, laboratory diagnosis and treatment are discussed. The need for future experimental studies is emphasized.