Effects of increased testosterone secretion on the behavior of adult male rhesus living in a social group. Selected members of an all-male social group of rhesus monkeys were treated with twice-weekly injections of human chorionic gonadotropin (HCG) to determine the effects of the consequently increased androgen secretion on social behavior. Radioimmunoassay of blood samples confirmed significant increases in serum testosterone levels in each of 4 subject, but only 3 of these showed significant behavioral changes. The basic social structure of the group, including the dominance hierarchy, was not disrupted by the treatment with HCG. Rather, the behavioral changes which did not occur appeared to be intensifications of previously existing social relationship.


Olfactory demarcation of territorial boundaries by a primate--Propithecus verreauxi. Field observations of a troop of sifaka revealed that they scent mark the boundary areas of their territory far more than they do the interior, both during normal daily movements and during battles between adjacent troops. They are active throughout the territory but spend more time and have a higher frequency of marking at the periphery.


Backward and forward masking in Macaca mulatta. 8 highly trained rhesus monkeys were examined on both backward and forward masking to determine the feasility of studying masking in unrestrained monkeys and of studying perceptual phenomenon without the confounding of linguistic variables. It was found that monkeys could respond well under the given conditions. The data indicated a more pronounced effect of the masking stimulus on backward masking than on forward masking.


Influence of sociality following isolation on feeding rhythms of monkeys entrained to artificial light schedules. Socialization following a period of isolation disrupted the feeding rhythms of monkeys entrained to artificial light schedules. When isolation was reinstituted, the performances of the subjects, which had shifted during socialization, reverted to profiles similar to but not the same as they were in the initial isolation period. The strength of the social factor may have derived, in part, from the effects of the isolation which preceded the socialization phase.


Troop male membership changes and infant killing in langurs (Presbytis entellus). Almost all studies of Presbytis entellus have resulted in observations of troop male membership changes. Reports from three studies indicate that in some cases troop male membership change occurs through rapid and complete adult male replacement with attendant infant mortality. Data from other investigations of P. entellus show or suggest a different pattern of troop male membership change with no infant mortality. The phenomena of infant killing and rapid male replacement are evaluated on the basis of review of data from ten P. entellus studies. While these and my own data indicate that male social instability (i.e. frequent changes in troop male membership) is species-typical for P. entellus, they do not support the hypothesis that infant killing represents part of an evolved male reproductive strategy.


Locomotion, limb proportions, and skeletal allometry in lemurs and lorises. The allometric patterns of postcranial scaling in lemurs (Lemuridae and Indriidae) and lorises (Lorisinae) are established and discussed in relation to locomotor and postural adaptations. The scaling trends of each limb and its components vary significantly among these three prosimian groups, but all are functionally explicable in reference to habitual locomotor behavior and details of substrate utilization.


[Hemoglobins, XXX. The amino acid sequence of the monomeric hemoglobin from Lampetra fluviatilis (author's transl)]. The primary structure of the two main hemoglobin components of the river lamprey Lampetra fluviatilis was established. Having a chain length of 149 amino acids the molecular weight of the globin is 16272. The two main components differ only by an N-formyl residue at the N-terminal proline. The sequence was compared with those of two related species. The heterogeneity of Cyclostomata hemoglobins and their possible origin was discussed.


[Rule of antibody structure. Primary structure of a human monoclonal IgAl-immunoglobulin (myeloma protein Tro). VI. Amino acid sequence of the L-chain, lambda-type, subgroup II]. The primary structure of the L-chain of an IgA1-immunoglobulin (Myeloma protein Tro) has been determined by means of cleavage with trypsin and, if necessary, with alpha-chymotrypsin. The tryptic peptides of the variable part were characterized by amino acid analysis, Dansyl-Edman degradation and cleavage with carboxypeptidase; the peptides of the constant part were identified by amino acid analyses and determination of its N- and C-terminal residues. The sequence of the remaining amino acids and the arrangement of the peptides were established in homology to known structures. The protein comprises 216 amino acids. The homology of the variable part clearly characterizes it as belonging to subgroup II of lambda-chains. In positions 27a, b and c, there are the subgroup-specific additional residues and in position 96 is the characteristic deletion. The constant part of the chain is Kern- and Oz- which indicates that it has serine in position 154 and arginine in position 191.


Respiratory syncytial virus infection in owl monkeys: viral shedding, immunological response, and associated illness caused by wild-type virus and two temperature-sensitive mutants. Intranasal inoculation of owl monkeys with wild-type respiratory syncytial virus induced upper respiratory tract disease in each of seven animals. The response of owl monkeys to two highly defective, temperature-sensitive, multiple-lesion mutants was then compared to the pattern seen with wild-type respiratory syncytial virus. These mutants, ts-1 NG-1 and ts-1 NG-16, were derived from the ts-1 mutant that had been remutagenized with nitrosoguanidine (NG). Previously the ts-1 NG-1 and ts-1 NG-16 mutants had been shown to be more temperature sensitive and more stable genetically than their ts-1 parent. Both ts-1 NG-1 and ts-1 NG-16 produced infection that was delayed in onsent compared to wild-type virus infection. However, the mutants were shed from the upper respiratory tract for the same period of time and at the same titer as wild-type virus. The serum neutralizing antibody response to infection with the mutants was nearly equivalent to that elicited by wild-type virus. However, the extent of disease induced by the mutants was significantly less than that seen with wild-type virus. These observations suggest that the mutants are potential vaccine condidates and should be subjected to additional in vivo testing in primates and, ultimately, humans.


Purification and characterization of a hemolysin produced by Vibrio cholerae biotype El Tor: another toxic substance produced by cholera vibrios. A thermolabile direct hemolysin from an El Tor cholera vibrio strain has been isolated and partially characterized as a simple protein of ca. 20,000 molecular weight. In addition to its hemolytic activity, the hemolysin is cytotoxic, cardiotoxic, and rapidly lethal. In these respects it resembles the thermostable direct hemolysin/cytotoxin/cardiotoxin/lethal toxin of Vibrio parahaemolyticus and certain other bacterial hemolysins, although there are other significant differences. Because identical diseases are produced by both hemolytic and nonhemolytic cholera vibrios, the El Tor hemolysin may be presumed to be pathogenetically irrelevant. These observations raise the question of "When is a toxic substance also a toxin?"


Effect of metal ions on diphtheria toxin production. The effect of several metal ions on the production of diphtheria toxin was tested. By using the gel immunodiffusion system for detecting toxin, a wide range of metal ion concentrations was conveniently surveyed. Five divalent cations, Fe2+, Cu2+, Co2+, Ni2+, and Mn2+ inhibited toxin production within a range of concentrations that did not inhibit growth of the producing strain. Growth and toxin production were inhibited at identical concentrations by both Cd2+ and Zn2+, whereas Al3+ and Sr2+ affected neither growth nor toxin production over the range of concentrations tested. The data showed that Fe2+ was the most effective inhibitor on an equivalence basis, followed by Cu2+, Co2+, and Ni2+ in descending order. All eight strains of Corynebacterium diphtheriae chosen from diverse ecological origins responded similarly to all metals at similar concentrations. A mutant strain which produces toxin at Fe2+ concentrations 500 times greater than are inhibitory for the parent strain had simultaneously acquired resistance to inhibitory concentrations of Cu2+, Co2+, Ni2+, and Mn2+. This suggests that there is at least one common point in the activity of all these metal ions, and that toxin may respond broadly to changes in metal ion concentrations in the environment.


Experimental infection of the genital tract of female grivet monkeys by Mycoplasma hominis: effects of different routes of infection. In a previous study we produced acute salpingitis and parametritis in grivet monkeys by inoculation of Mycoplasma hominis directly into the uterine tubes. With the purpose of examining in the same animal model the effect of more natural routes of infection, six female grivet monkeys were inoculated, two by two, with M. hominis by the following methods: (experiment A) into the uterine cavity through the cervical canal; (experiment B) into the uterine cavity by the same route, but after ligature of the isthmus of the uterine tubes had been performed and followed by curettage of the endometrium; and (experiment C) into the cervical epithelium. Whereas the animals in experiment A developed no or only very slight signs of infection, all monkeys in experiments B and C developed pronounced gross and microscopic inflammatory lesions of the uterine tubes and parametria, together with a significant antibody response. It is concluded that ascending M. hominis infection of the genital tract must be preceded by mechanical injury of the epithelial barrier, and that subsequent spread of the infection occurs via blood and lymph vessels rather than by the canalicular route. The inflammatory genital tract disease produced experimentally with M. hominis in grivet monkeys is very similar in its pathogenesis and pathology to the type of salpingitis caused in humans by microorganisms other than Neisseria gonorrhoeae and Chlamydia trachomatis.


Alternate complement pathway activation by group A streptococci: role of M-protein. Avirulent strains of group A streptococci readily activate the complement system in normal human serum via the alternate complement pathway (ACP). Virulent M-positive group A streptococci are much less potent as activators of the ACP. The ability of M-positive streptococci to activate the ACP is enhanced by trypsinization or mild peptic digestion. The latter treatment removes the serologically active and antiphagocytic type-specific moieties of M protein, but retains the surface fuzzy layer. The phagocytosis of avirulent streptococci is markedly enhanced by preopsonization in serum chelated with Mg-ethylene glycol tetraacetic acid (classic complement pathway blocked) but not in serum devoid of heat-labile factors. These studies suggest that the function of M protein as a virulence factor may be mediated, at least in part, by its ability to retard interaction of ACP components with structures present on the streptococcal cell surface.


Antigens of Streptococcus mutans: cellular localization of the serotype-specific polysaccharide of strain AHT and release during exponential growth. The serotype-specific polysaccharide of Streptococcus mutans AHT (serotype a) was shown to be loosely associated with the cell surface of this organism. The antigen was extracted from whole cells by boiling in sodium acetate buffer, pH 4.0, for 10 min. The purified product was found to be a diheteroglycan of galactose and glucose (3.6:1, molar ratio). The antigen possessed serological characteristics similar to the a antigen previously extracted from purified cell walls with hot formamide. Its physicochemical structure was identical to the previously studied wall antigen. Electron micrographs, developed after immunocytological labeling of this antigen on whole cells, revealed it to compose a dense microcapsule surrounding the microbe. Analyses of spent culture fluids indicated that the antigen was released during exponential growth at a rate directly proportional to the increase in culture biomass. It is concluded that the serotype-specific antigen may be a prime immunogen due to its surface localization at both capsule and wall sites.


Partial purification of a specific antigen of Treponema hyodysenteriae. A specific antigen of pathogenic Treponema hyodysenteriae was extracted from lyophilized cells of isolant B169 with hot phenol-water, isolated, and semipurified by starch block electrophoresis.


Immunity in Schistosoma mansoni using antigens of Fasciola hepatica isolated by concanavalin A affinity chromatography. Antigens of Fasciola hepatica adult worms were chromatographed using concanavalin A-Sepharose 4B. Two unbound peaks appeared in the inclusion volume (DT-1 and DT-2), and one peak was eluted with alpha-methylglucoside (E1-1). At least seven peaks were obtained by isoelectric focusing of E1-1. The largest of these peaks, with an average pI of 4.0, contained the antigens reactive with antibodies to Schistosoma mansoni. Mice immunized with DT-2 or E1-1 and challenged with S. mansoni cercariae developed 39 to 82% fewer worms than controls. DT-1 had no protective effect. Combining DT-1 and DT-2 abolished this protection. These experiments demonstrate that F. hepatica glycoprotein antigens induce in mice significant protection to infection with S. mansoni and offer an interesting approach to the study of vaccines in experimental schistosomiasis.


Suppression of antibody response to Leptospira biflexa and Brucella abortus and recovery from immunosuppression after Berenil treatment. Zebu cattle infected with either Trypanosoma congolense EATRO 1800 or Trypanosoma vivax EATRO 1721 had suppressed humoral immune responses to Leptospira biflexa injected intravenously and to attenuated Brucella abortus injected subcutaneously. T. congolense infections were more suppressive than T. vivax infections. In cattle infected with T. vivax, the suppression of immune responses to both bacterial immunogens was abrogated when the animals were treated with Berenil at the time of antigen administration. In cattle infected with T. congolense, simultaneous Berenil treatment at the time of vaccination abolished the suppression of immune response to L. biflexa, and lessened but did not abrogate the suppression of immune response to B. abortus.


Comparison of two antigen detection techniques in a primate model of Haemophilus influenzae type b infection. Rapid diagnosis of Haemophilus influenzae type b meningitis is possible using immunological tests for capsular antigen (polyribophosphate, PRP), such as countercurrent immunoelectrophoresis (CIE) and latex particle agglutination (LPA). We compared two tests in monkeys with evolving, serially quantitated H. influenzae type b bacteremia (n = 23) and meningitis (n = 21). In vitro, the LPA test was sensitive to 0.5 ng of PRP/ml of saline, and the CIE test was sensitive to 1.0 ng/ml; in serum, however, CIE detected 5.0 ng of PRP/ml, whereas the sensitivity of LPA was unchanged. LPA detected PRP earlier in the course of bacteremia (mean, 12 h after onset; range, 4 to 36 h) than did CIE (mean, 45 h; range, 4 to 168 h) (P less than 0.01). A positive LPA test required greater than or equal to 100 bacteria per ml of blood, whereas CIE required greater than or equal to 1,000/ml. PRP accumulated with continuing blood stream infection, aiding detection of low-grade bacteremia. LPA detected antigen in cerebrospinal fluid (CSF) earlier in the course of meningitis and at a lower bacteria density than did CIE. Both methods detected antigen reliably with greater than or equal to 1,000 bacteria per ml of CSF. A close correlation existed between CSF concentrations of capsular antigen and bacteria (r = 0.90, P less than 0.001). We conclude that the LPA method permits earlier diagnosis of H. influenzae type b infection in part because of its greater sensitivity.


Enzymatic induction of interferon production by galactose oxidase treatment of human lymphoid cells. Human lymphocyte cultures produced large amounts of interferon after treatment with the enzyme galactose oxidase. Interferon production was detectable as early as 3 h after enzymatic treatment and reached a level of about 10(4) reference units 20 to 24 h later. Galactose oxidase-induced interferon appeared to be immune interferon on the basis of acid lability, lack of neutralization by antibody to leukocyte interferon, and slow kinetics of activation of the cellular antiviral state. Interferon production was inhibited to the same extent (99%) by pretreatment of the cells with beta-galactosidase or with neuraminidase followed by beta-galactosidase, suggesting that the critical event for activation of interferon production is the oxidation of exposed galactose residues on lymphocyte membrane.


Leishmanial excreted factor: protein-bound and free forms from promastigote cultures of Leishmania tropica and Leishmania donovani. Leishmania spp. growing in culture produce an immunologically active substance called excreted factor (EF), which precipitates antibodies raised against intact cells and has been implicated as the conditioning agent for parasite infection of host macrophages. An improved method for isolation of the material is described, based on Sephadex column chromatography of growth medium which had been boiled at pH 5.0. This procedure allows the detection of differences among the EF molecules of different species, and it overcomes previous shortcomings through the monitoring of immunological activity throughout. Analysis of the products of this procedure revealed that EFs from Leishmania tropica and Leishmania donovani share a common carrier protein, identified as rabbit serum albumin, and are chemically quite similar. Growth medium from L. tropica boiled at acidic pH contains primarily an EF-albumin complex of 75,000 molecular weight. Treated growth medium from L. donovani, on the other hand, contains both the albumin complex and a smaller molecule (less than 27,000 molecular weight) that is not associated with rabbit protein. This material accounts for nearly 20% of the EF of one L. donovani strain, but constitutes only a minute fraction of L. tropica EF. Treatment of the EF-albumin complex with trichloroacetic acid separates the molecule into two major subunits, one having a molecular weight of about 61,000 (without anti-Leishmania activity) and the other having a molecular weight of about 18,000 (with no anti-rabbit activity). The protein-free EF of L. tropica differs from that released by trichloroacetic acid extraction in that it is capable of precipitating antisera of nonhomologous serotypes, whereas the albumin complex and the trichloroacetic acid-treated EF fragment are not. EFs from both species display pH-dependent affinity for certain lectins.


Light chain isotype-associated suppression of normal plasma cell numbers in patients with multiple myeloma: Medical Research Council's Working Party for Leukaemia in Adults and the Oxford Lymphoma Group. The number of plasma cells in the lamina propria of the gut has been assessed in patients with multiple myeloma and other B-cell neoplasms. The total number of these plasma cells was reduced in most patients with myelomatosis and one-third of patients with lymphoplasmacytoid tumours. This reduction was not, however, seen in patients with other neoplasms of B-cell origin, although hypogammaglobulinaemia was common to all groups of patients. The depletion of gut plasma cell numbers was not uniform in myelomatosis patients. They showed selective loss of plasma cells with the same light chain isotype as that produced by the neoplastic clone.


Occurrence of heat-labile Regan type of alkaline phosphatase in hematopoietic tumors. The physiocochemical and immunological properties of alkaline phosphatase extracted from Hodgkin's nodes, non-Hodgkin's lymphoma nodes and leukemic leukocytes have been studied. The alkaline phosphatase from these three tumor types possesses the same biophysical and biochemical properties and immunological determinants as the placental alkaline phosphatase. However, it is more heat-labile than the placental isoenzyme. Immunological experiments indicate that, of these tumor types, Hodgkin's tumor contains the largest amounts of heat-labile Regan type of alkaline phosphatase.


The initiation of tumours on mouse skin by dihydrodiols derived from 7,12-dimethylbenz(a)anthracene and 3-methylcholanthrene. The cis-2a,3-diol and the trans-4,5-, trans-7,8-, trans-9,10- and trans-11,12-dihydrodiols of 3-methylcholanthrene and the trans-3,4, trans-5,6-, trans-8,9. and trans-10,11- dihydrodiols  of 7,12-dimethylbenz[a]anthrancene have been tested, in comparison with the parent hydrocarbons, for their abilities to initiate skin tumours in female CDI mice. Groups of mice received a single topical application (25 micrograms) of a diol or of a hydrocarbon, and 1 week later repeated topical applications (1 microgram) of 12-0-tetradecanoylphorbol-13-acetate were commenced. The results show that the diol of 3-methylcholanthrene and the 3,4-diol of 7,12-dimethylbenz[a]anthrancene were active as initiating agents but that they were no more active than their parent hydrocarbon. The K-region 5,6-diol of 7,12-dimethylbenz[a]anthrancene, which cannot be converted directly into a vicinal diol-epoxide, was also active as a tumour-initiating agent when applied to mouse skin.


Clinical pharmacokinetics of nitroxoline. 14C-Nitroxoline was given orally to the rats, and its distribution as well as plasma and bile levels were determined autoradiographically and by the aid of radioactivity measurements, respectively. Nitroxoline was also given to the human volunteers orally and intravenously in three various doses and the corresponding urine concentrations of unconjugated and conjugated nitroxoline were determined spectrophotometrically. A pharmacokinetical model was generated on the basis of the results. The curve fitting procedure between total nitroxoline cumulative quantities in urine and the model response simulated on analog-hybrid computer enabled the evaluation of the validity of the chosen model as well as of the identification of its parameters.


The influence of caloric restriction on serum prolactin. Nine obese female patients were placed on a liquid formula diet containing 320 kcal (1.34 MJ), 31 g protein, 44 g carbohydrate, 1.5 g fat, vitamins and essential minerals for a period of four weeks under metabolic-ward conditions. The diet was tolerated well in all patients and mean weight loss was 2.49 kg per week during the four-week period. Basal serum prolactin levels were normal (2.9 +/- 0.6 ng/ml) and did not show any change during this period. In six patients a TRH test was performed before and during caloric restriction. The maximal increment of PRL after TRH was significantly lowered during dietary treatment: 53.2 +/- 16.2 vs. 32.3 +/- 11.6 ng/ml; mean +/- s.e.m.; P less than 0.025). No change was observed in the serum TSH responset to TRH before and during caloric restriction. Dietary restriction suppresses prolactin synthesis and secretion as measured in response to TRH in obese female patients.


The role of liver passage for conversion of pteroylmonoglutamate and pteroyltriglutamate to active folate coenzyme. In an attempt to clarify the possible site of conversion of folates to active methylated coenzyme forms mono- and triglutamate was injected rapidly into the portal vein of dogs. Samples of blood were removed at various intervals from catheters placed in the portal vein, the hepatic vein and the femoral artery for assay carried out both with L. casei, S. Faecalis and liquid scintillating counting. Unreduced monoglutamates were extracted in the liver, immediately converted to methyltetrahydrofolate and released again to plasma from which it was slowly taken up by peripheral tissue. Reduced folates were refused by the liver. Triglutamates were extracted exclusively in the systemic circulation, converted to unreduced monoglutamate later to be reduced in the liver. No tissue displacement of folate was demonstrated. Only small amounts of monoglutamates, mainly reduced forms, were lost in the urine during the experiments. The kinetics of unreduced monoglutamate suggest a one compartment system, while the kinetics of methyltetrahydrofolate and triglutamate may implicate a two compartment system.


Adenosine deaminase activity of normal lymphocytes and leukemic cells. Adenosine deaminase (ADA) activity was determined in peripheral blood T cells, thymocytes and blasts of 42 acute leukemia patients. Thymocytes had higher ADA activity than mature peripheral blood T cells. B ALL (acute lymphoblastic leukemia) cells had significantly lower activity compared with T ALL and non-B, non-T cells. The latter had a wide range of enzymatic activities but mostly in the T cell range. Based on these results, as well as on assays of B and T cell lines, it seems that thymus-derived cells--and especially prethymic and immature T cells--have high ADA activity. This finding could be useful in subtyping the heterogeneous group of non-B, non-T ALL.


Biotypes of Brucella abortus and their value in epidemiologic studies of infected cattle herds. Four Texas cattle herds containing cows infected with either Brucella abortus biotype 1, 2, or 4 were studied to determine the probability of transmission of Brucella between adjacent cattle herds, the most probable means by which Brucella was introduced into the herds, and the relative frequency of strain 19 isolation from vaccinated cattle. A total of 1,935 cattle in the four herds were tested for brucellosis; 339 reactors were identified, and isolations of B abortus were made from 143. The biotype of B abortus was used to determine that purchased cattle or reentry of bred heifers into the herds was probably responsible for introducing B abortus and that the biotype was not readily transmitted to adjacent herds. Three (9%) of 32 B abortus isolations from adult-vaccinated cattle were strain 19. The data supported the hypothesis that biotypes can be useful in determining the source of B abortus for cattle and in differentiating field and vaccine strain infections in adult-vaccinated cattle.


Induction of resistance to aurodox by aurodox in the antibiotic-producing culture, Streptomyces goldiniensis. The sensitivity of protein and aurodox synthesis to aurodox was examined in relationship to the development of resistance to aurodix on Streptomyces goldiniensis during fermentation. It was found that the culture remains sensitive to the antibiotic as long as no aurodox is present in the medium. Resistance only develops when aurodox is present, either exogenously added or endogenously synthesized by the culture. These observations suggest that the development of resistance is an inducible process, and evidence is presented indicating that aurodox induces a specific resistance system in S. goldiniensis.


Studies on bacterial cell wall inhibitors. VIII. Mode of action of a new antibiotic, azureomycin B, in Bacillus cereus T. Azureomycin B, a new antibiotic which contains sugar, amino acid and phenol moieties and inhibits Gram-positive bacteria, was found to be a specific inhibitor of peptidoglycan synthesis in bacteria. The antibiotic lysed growing cells of Bacillus cereus T at a concentration of 10 micrograms/ml but did not affect resting cells. Microscopical observation revealed swelling and lysis of the bacterial rods when treated with azureomycin B. The incorporation of [3H]diaminopimelic acid or [14C]glucosamine into acid-insoluble fraction of growing cells of Bacillus cereus T was strongly inhibited in the presence of azureomycin B, but that of [14C]leucine, [3H]thymidine or [3H]uridine were not, at least until 5 minutes after the beginning of the incubation. The antibiotic caused accumulation of a nucleotide precursor in the cells which was identified as UDP-MurNAc-L-Ala-D-Glu-meso-Dpm-D-Ala-D-Ala. Thus the site of action was suggested to lie between this nucleotide and peptidoglycan in the pathway of peptidoglycan synthesis.


Controlled biosynthesis of neoviridogriseins, new homologues of viridogrisein. IV. In vitro synergism between neoviridogrisein II and the antibiotics of the mikamycin A group. Neoviridogrisein II is a homologue of viridogrisein in which the hydroxyproline residue is replaced by proline. Neoviridogrisein II proved to be more active than the parent antibiotic against Gram-positive bacteria and Mycoplasma species. When neoviridogrisein II or viridogrisein was combined with griseoviridin, a non-peptidyl macrocyclic lactone, synergism was observed: maximum synergistic effect was observed for a combination ratio that depended on the test bacterium used. Neoviridogrisein II also exerted synergism when combined with mikamycin A and A-2315A.


In vitro and in vivo laboratory studies on three hydroxyiminophenylacetyl cephalosporins with particular reference to SK&F 80303, an unusually long-acting cephalosporin. Three cephalosporins with 7-(2-hydroxyiminophenylacetamido) side chains (SK&F 79433, 80000 and 80303), differing in their 3-substituents, exhibited similar broad-spectrum antibacterial activity in vitro against strains of Staphylococcus aureus, Streptococcus faecalis and various Gram-negative bacilli. All three were active in vivo (s.c., mouse) against S. aureus, Escherichia coli or Klebsiella pneumoniae, but they differed significantly in serum pharmacokinetic profiles. SK&F 80303 produced high and extremely prolonged serum levels and protected mice when administered up to 24 hours prior to challenge with beta-lactamase-producing S. aureus or K. pneumoniae. It was resistant to hydrolysis by beta-lactamases from S. aureus, and variably so to beta-lactamases from E. coli strains. SK&F 80303 was bacteriolytic to logarithmically growing S. aureus, E. coli, Proteus mirabilis, K. pneumoniae and Enterobacter cloacae (partially). SK&F 80303 illustrates further the effect of the 3-sulfoalkyltetrazole substituent on the pharmacokinetic properties of cephalosporins. Its combined biological properties make it a possible candidate for therapeutic and long-term prophylactic use.


Comparative bactericidal activities of beta-lactam antibiotics determined in agar and broth media. Comparative bactericidal activities were determined utilizing a relatively large number of test strains, in both agar and broth media, with special reference to the time of exposure of the bacteria to certain beta-lactam antibiotics. It was apparent that the activities increase with time. The concentrations producing a 99.9% kill with cephalothin for Escherichia coli, Klebsiella sp., and carbenicillin for Pseudomonas aeruginosa were higher in broth than in agar. In contrast, those of benzylpenicillin for alpha-streptococcus (non-enterococcal) were higher in agar than in broth. If the bactericidal concentrations with 3-hour or 6-hour exposure to antibiotics were used as the criterion, these concentrations of carbenicillin for P. aeruginosa, and benzylpenicillin for alpha-streptococcus were, in particular, unusually high compared with the conventionally determined bacteriostatic concentrations (MICs).


Disodium D-6-[alpha-(1,2,4-triazine-3,5-dione-6-carboxamido)-4-hydroxyphenylacetamido]penicillanate, BL-P1908. The synthesis of a new penicillin, disodium D-6-[alpha-(1,2,4-triazine-3,5-dione-6-carboxyamido)-4-hydroxyphenylacetamido]penicillanate (BL-P1908), is described. The compound shows superior in vitro activity against Pseudomonas aeruginosa compared to carbenicillin and ticarcillin and produces higher intramuscular serum levels in mice than does carbenicillin.


Immunological cross-reactivities of sulfocephalosporins. The immunological cross-reaction among sulfocephalosporins (cefsulodin, SFC-I, SFC-II), cephaloridine, sulbenicillin, penicillin G and 7-aminocephalosporanic acid has been investigated. A specific antibody was produced in rabbits against several structurally related haptens. The sensitization was conducted with a hapten-human serum albumin conjugate, which was prepared in an alkaline solution, with Freund's complete adjuvant. A cross-reaction was claraly demonstrated between sulfocephalosporins and sulbenicillin, but not between sulfocephalosporins and cephaloridine, penicillin G or 7-aminocephalosporanic acid.


Aflatoxin variation among corn samples with varying ratios of Aspergillus flavus-inoculated/noninoculated kernels. Aflatoxin levels and physical properties of corn kernels inoculated with Aspergillus flavus during development and noninoculated kernels were compared in samples with various proportions of the 2 kernel types. The relationship between mean toxin levels and associated standard deviations of 5 samples demonstrated a linear association from the lowest toxin in noninoculated corn through a mixture of 60% inoculated/40% noninoculated. However, at the highest toxin level in the 100% inoculated material, a reduction in sample variation was observed. Examination of individual kernal weights showed that inoculated kernels were distinctly lighter than noninoculated seed. A uniform grinding procedure of the samples yielded heterogeneous particle sizes based on the starting corn. The large particle fraction (greater than 500 micrometers) decreased from 100% noninoculated kernels through the mixtures to the 100% inoculated seed; particles below 150 micrometers were most abundant in the ground samples from inoculated kernels. In addition, the density of particles within a size category varied; lower densities were observed in samples obtained from A. flavus-inoculated kernels.


Reverse phase ion pair high pressure liquid chromatographic determination of ethylenediaminetetraacetic acid in crabmeat and mayonnaise. A method is described for the determination of ethylenediaminetetraacetic acid (EDTA) in crabmeat and mayonnaise. EDTA is extracted from the food sample with water and converted to its copper chelate, which is then quantitated by reverse phase ion pair high pressure liquid chromatography with ultraviolet detection. Maximum sensitivity is obtained with detection at about 254 nm; higher wavelengths may be used for enhanced specificity. Cleanup procedures for crabmeat and mayonnaise were improved by using a radiotracer method. Analyses of crabmeat and mayonnaise samples spiked at 3 different levels showed greater than 90% recovery of EDTA.


Equivalence of microbiological and hydroxylamine methods of analysis for ampicillin dosage forms. Ampicillin formulations were assayed by microbiological and hydroxylamine methods to determine whether thehydroxylamine analytical method is a suitable substitute for the microbiological method. Paired assay results by the 2 analytical methods were obtained on different strengths of tablet, capsule, and suspension, formulations containing ampicillin and ampicillin degradation compounds. Several statistical tests were used to assess the equivalence of the paired assay results. The data analyses indicate that the hydroxylamine method is a suitable substitute for the microbiological method for potency assays and stability studies of ampicillin formulations. The hydroxylamine method yielded slightly higher assay results than the microbiological method for severely degraded formulations.


Reconstituted O-dealkylase systems containing various forms of liver microsomal cytochrome P-450. Among the seven forms of purified liver microsomal cytochrome P-450 tested, P-4501, P-4502 (both from phenobarbital-treated rabbits), P-4504 (from phenobarbital-treated rats), and P-4482 (from methylcholanthrene-treated rats) could reconstitute significant activities catalyzing the O-deethylation of 7-ethoxycourmarin, but remarkable differences were seen among the catalytic properties of the four reconstituted systems. The systems containing P-4501, and P-4504 required the presence of cytochrome b5 for maximal activity, but no such requirement was observed with the other two systems. The Km value of the P-4502 system for ethoxycoumarin was of the order of 10(-7) M, whereas those of the other systems were about 1,000 times higher. The Vmax value determined for the P-4482 system was higher than those for the other systems by a factor of 10. 7-Methoxycoumarin was metabolized in a way similar to ethoxycoumarin by the systems containing P-4501, P-4502, and P-4504, but acted as a strong uncoupler in the P-4482-containing system. In the P-4482 system, however, ethoxycoumarin O-deethylation was almost completely coupled to NADPH oxidation. In the other systems, on the other hand, NADPH oxidation was partially uncoupled to similar extents with respect to the product formation with both ethoxy- and methoxycoumarins as substrates. The four systems could also be distinguished from one another with respect to the effects of several inhibitors. The P-4502-containing system, but not the other three systems, was progressively inactivated during methoxycoumarin O-demethylation and benzphetamine N-demethylation. Such inactivation was not observable during the ethoxycoumarin O-deethylation reaction. It is suggested that the active site of P-4502 reacted with formaldehyde produced by the demethylation reactions and was thus irreversibly inactivated. The results reported in this paper provide a clear example of different catalytic properties of multiple forms of hepatic microsomal cytochrome P-450.


Comparison of the 95,000 molecular weight protein from Limulus sperm with muscle alpha-actinin. The 95,000 molecular weight protein (95K protein) of the false discharges of Limulus sperm, purified by means of preparative gel electrophoresis in the presence of sodium dodecyl sulfate, was compared with a 95K protein from Limulus muscle and chicken gizzard alpha-actinin. The results were as follows. 1) One-dimensional peptide mapping using four different proteases showed differences among these proteins. 2) Two-dimensional peptide mapping using trypsin showed that about 30% of the peptides in the digest of the sperm 95K protein were similar to those of chicken gizzard alpha-actinin and about 50% of the peptides were similar to those of the Limulus muscle 95K protein. 3) The sperm 95K protein contained relatively large amounts of Gly, Pro, and Ser and relatively small amounts of Glu and Leu compared to the muscle proteins. 4) Antibodies against the sperm 95K protein did not cross-react with the Limulus muscle 95K protein or chicken gizzard alpha-actinin. These results suggest that the 95K protein of sperm is different from alpha-actinin in primary structure.


Isolation and characterization of antithrombin III from human, porcine and rabbit plasma, and rat serum. 1. Human, porcine, rabbit, and rat antithrombin III have been purified by affinity chromatography using heparin-agarose. The amino acid and carbohydrate compositions, amino-terminal sequences, immunological cross-reactivities, and inhibitions of human thrombin were studied. 2. Human, porcine, rabbit, and rat antithrombin III are single-chain glycoproteins containing hexose, glucosamine, and neuraminic acid. 3. The total carbohydrate contents were 17, 16, 14, and 15% for human, porcine, rabbit, and rat antithrombin III, respectively. 4. Molecular weights estimated from the migration in sodium dodecyl sulfate (SDS)-poly-acrylamide gel electrophoresis were 59,000, 58,000, 63,000, and 63,000 for human, porcine rabbit, and rat antithrombin III, respectively. 5. These four proteins have similar amino acid compositions, although some minor differences were noted. 6. Human, porcine, and rabbit antithrombin III have a histidine residue at the amino-terminus, while rat antithrombin III contains an amino-terminal asparagine residue. 7. The amino-terminal sequences up to the first 17 residues showed high homology among the four proteins. 8. Some immunological cross-reactivity was observed only between human and porcine antithrombin III. 9. The apparent dissociation constants (KI) for the complexes between human thrombin and human, porcine, rabbit, and rat antithrombin III were about 1.2 x 10(-10) M, 9.5 X 10 (-9) M, 1.4 X 10(-7) M, and 2.8 X 10(-9) M, respectively.


Postnatal development of peroxisomal and mitochondrial enzymes in rat liver. Subcellular organellles from livers of rats three days prenatal to 50 weeks postnatal were separated on sucrose gradients. The peroxisomes had a constant density of 1.243 g/ml throughout the life of the animal. The density of the mitochondria changed from about 1.236 g/ml at birth to a constant value of 1.200 g/ml after two weeks. The peroxisomal and mitochondrial fatty acid beta-oxidation and the peroxisomal and supernatant activities of catalase and glycerol-3-phosphate dehydrogenase were measured at each age, as well as the peroxisomal core enzyme, urate oxidase, and the mitochondrial matrix enzyme, glutamate dehydrogenase. All of these activities were very low or undetectable before birth. Mitochondrial glutamate dehydrogenase and peroxisomal urate oxidase reached maximal activities per g of liver at two and five weeks of age, respectively. Fatty acid beta-oxidation in both peroxisomes and mitochondria and peroxisomal glycerol-3-phosphate dehydrogenase exhibited maximum activities per g of liver between one and two weeks of age before weaning and then decreased to steady state levels in the adult. Peroxisomal beta-oxidation accounted for at least 10% of the total beta-oxidation activity in the young rat liver, but became 30% of the total in the liver of the adult female and 20% in the adult male due to a decrease in mitochondrial beta-oxidation after two weeks of age. The greatest change in beta-oxidation was in the mitochondrial fraction rather than in the peroxisomes. At two weeks of age, four times as much beta-oxidation activity was in the mitochondria as in the peroxisomal fraction. Peroxisomal glycerol-3-phosphate dehydrogenase activity accounted for 5% to 7% of the total activity in animals younger than one week, but only 1% to 2% in animals older than one week. Up to three weeks of age, 85% to 90% of the liver catalase was recovered in the peroxisomes. The activity of peroxisomal catalase per g of rat liver remained constant after three weeks of age, but the total activity of catalase further increased 2.5- to 3-fold, and all of the increased activity was in the supernatant fraction.


Potentiation by p-fluorophenylalanine of the division-delaying effects of high hydrostatic pressure in synchronized Tetrahymena pyriformis. Cultures of heat-synchronized Tetrahymena pyriformis, growing in a proteose peptone medium, were subjected to short pulses of the amino acid analogue, p-fluorophenylalanine, and high hydrostatic pressure. The pulses of these agents were chosen so that, when applied individually, they did not appreciably delay cell division. However, combined treatments, analogue pulse followed by pressure pulse, produced a pronounced synergism. The results are interpreted as further evidence to support the inclusion of analogue division proteins in pressure labile assemblies during the progression of Tetrahymena into division.


DNA synthesis: evaluation of a hydrodynamic method for measuring the rate of replication fork movement. When replicating DNA is labeled sequentially with radioactive and density tracers and analyzed by equilibrium centrifugation, the fraction banding at heavier than normal density is inversely proportional to the rate of replication fork movement if there is a sharp transition from one tracer to the other on the newly synthesized chains (Painter and Schaefer, '69). Primate CV-1 DNA labeled for 5 to 30 minutes with 3H-dThd and then for three hours with BrdUrd in the presence of FdUrd bands in a bimodal distribution in alkaline CsCl, rather than in a continuous distribution with a skew toward heavier density seen when FdUrd is omitted and centrifugation is in neutral CsCl. The heavy density peak represents interspersion of both tracers in the DNA and is caused by slow transition from dThd to BrdUrd incorporation when the tracers are switched in the labeling medium. This may result from preferential uptake and incorporation of dThd over BrdUrd. Because of the interspersion, calculation of the rate of replication fork movement is inaccurate. Reversal of the labeling sequence with administration of the long density pulse before the radioactive pulse reduces the problem of interspersion. Using this sequence of labeling, estimates of the rate of fork movement of 0.36-0.38 micrometer/min are obtained when the 3H pulse time is long enough to allow accurate measurement of the fraction of heavy DNA. Analysis by fiber autoradiography yields a rate of 0.56 micrometer/min in the same cell line. If appropriate precautions are taken to minimize mixing of the two tracers in the precursor pool and to ensure that the fraction of heavy DNA is measured accurately, the hydrodynamic technique provides an objective method of measuring rate of fork movement that gives values only slightly lower than those obtained by autoradiography.


Rapid alterations in cellular morphology and plasma membrane structure induced in rat thymocytes by mitogenic stimuli. Under conditions where a maximum stimulation of 3-O-methyl-glucose transport is observed, three thymocyte mitogens (concanavalin A, ionophore A23187 and hydrogen peroxide) cause cell rounding and a decrease in the density of intra-membrane particles on the plasma membrane. The early effects of mitogens on the thymocyte plasma membrane are similar to those of osmotic shock.


Co-operativity of lectin binding to fibroblasts and its relation to cellular actomyosin. We have investigated the binding of 125I-concanavalin A (125I-Con A) and 125I-succinyl concanavalin A (125I-s-Con A) to rat fibroblasts (16C line) as a function of the concentration of added lectin, and the alterations to this binding behaviour caused by drugs which modify the cytoskeleton. The changes in cell behaviour which occur at different levels of binding have also been studied. As shown previously for some other systems, the binding of Con A is complex and partly co-operative. Three phases can be distinguished in our system: (i) pre-nucleation binding, (ii) binding which shows a small positive slope in a Scatchard plot and a Hill coefficient greater than unity, and which therefore is incipiently co-operative, and (iii) post-co-operative binding. The co-operative phase of binding is paralleled by progressive inhibition of EGTA-mediated cell detachment from substrata, with inhibition being complete when this phase of binding is complete. Likewise, the phagocytosis of latex spheres is progressively inhibited up to a threshold which coincides with the completion of co-operative binding. Thirdly, cells pretreated with Con A round up with colchicine (10(-5) M) if co-operative finding is complete, but adopt broad epithelial shapes if it is not. s-Con A does not show cooperative binding, and correspondingly does not inhibit EGTA-mediated cell detachment, or show a distinct threshold in the inhibition of phagocytosis, or promote the 2 types of shape change with colchicine. The pattern of Con A binding is drastically altered by pretreatment of cells with cytochalasin B or azide. The Scatchard and Hill plots show that the co-operative phase remains and is complete at about the same level of binding, but that it is more readily nucleated and takes place against a changed number and/or distribution of receptors. Pretreatment of cells with colchicine causes changes in the pattern of binding which are different from those observed with cytochalasin B or azide and are more difficult to interpret. We conclude that a reciprocal relationship exists between the cellular actomyosin and the state of cell surface receptors. Perturbation of actomyosin by cytochalasin B or azide can enhance the freedom of some receptors to participate in a co-operative rearrangement which facilitates the binding of further molecules of lectin. Vice versa, the co-operative event has a feedback influence on the cellular actomyosin to cause alterations of cellular response.


Specific contact-dependent cell-to-cell communication during preconjugant interactions of the ciliate Euplotes crassus. A system has been developed to study cellular interactions between cells of complementary mating types prior to mating in the ciliate, Euplotes crassus. The presumptive mates were distinguished by using singlet and doublet cells of appropriate mating types in the mixtures. Cells of a given mating type were prelabelled with [3H]leucine and mixed with unlabelled complementary cells. Exchange of [3H]leucine-labelled material from donor to recipient cells was monitored through the various stages of the preconjugant interaction. A label transfer between the mating type complementary cells was detected from the beginning of the visible mating reaction, which occurs after a waiting period from the time of cell mixing and involves ciliary agglutination prior to cell body fusion. Complementary cells which were prevented from physically contacting each other and cells which were not competent to mate appeared unable to take up the labelled material. It is suggested that this material consists of some substance(s) playing an important role in the preconjugant cell-to-cell interactions of E. crassus.


Intracellular distribution of lead in Tetrahymena during continuous exposure to the metal. Lead acetate (0.1--0.2%) forms a precipitate with the organic growth medium. The Tetrahymena cells ingest this lead-containing precipitate and cell growth is resumed after a variable lag period. Ingested lead is observed as electron-dense material in food vacuoles. Soon after exposure, cytoplasmic lead (preserved with certain fixation only) is revealed as electron-dense particles in cilia and in a halo around digestive vacuoles. Later the lead particles pervade the entire cell but after the lag period they are confined to membrane-bound spaces. In dilute growth medium, high concentrations of lead inhibit food-vacuole formation and cell growth. Under these conditions lead is deposited in alveoli of the pellicle and is also found in autophagic vacuoles and other membrane-limited structures. The study has revealed that lead enters Tetrahymena through the membrane of digestive vacuoles and through the cell surface. The change in distribution of lead during the lag period indicates that a mechanism is activated for removal of lead into membrane-bound spaces. The final storage of lead seems to be in lysosomes.


High-performance liquid chromatographic analysis of biogenic amines in biological materials as o-phthalaldehyde derivatives. A remarkably sensitive, simple and selective reversed-phase high-performance liquid chromatographic (HPLC) method has been developed, allowing, for the first time, the direct measurement of histamine, norepinephrine, octopamine, normetanephrine, dopamine, serotonin and tyramine in a single sample of plasma (2 ml), tissue (0.2 g), or urine. The biogenic amines were modified by pre-column derivatization with o-phthalaldehyde which stabilizes the molecules, aids in extraction, and improves HPLC detection at the nanogram level. To minimize losses during the sampling procedure a careful collection procedure was designed. We developed a simple sample cleanup in which the samples were thawed, neutralized with KOH, immediately derivatized, extracted into ethyl acetate (EtOAc) and then chromatographed by HPLC. The derivatives were stable in EtOAc for more then 24 h. Interfering amino acids were removed from the EtOAc by partitioning twice with Na2HPO4 buffer (pH 10.0). Complete separation was achieved in ca. 60--90 min on a muBondapak phenyl column using a stepwise gradient of acetonitrile and/or methanol-phosphate buffer (pH 5.1). A variable wavelength fluorometer with a 5-microliter flow-cell was used (excitation 340 nm; emission 480 nm). Linearity ranged from 200 pg to 50 ng onto the column. Precision (R.S.D.) for retention times was 1% and for derivatization and injection 2.5%. Recoveries of the seven biogenic amines from plasma spiked with 25 ng/ml averaged 70%, with a relative standard deviation of 6%. Separation studies were also done using a muBondapak C18 column. The effects of various eluents are presented. Gas-liquid chromatography was also investigated but lacked the sensitivity achieved by HPLC. The HPLC method is used routinely for the determination of biogenic amines in plasma from pigs with malignant hyperthemia and thermally stressed bovine. Significant differences in levels of biogenic amines were noted between stressed and non-stressed animals. Data on rat brain tissue samples were compared with the trihydroxyindole method and canine heart tissue was analyzed for ventricular norepinephrine and dopamine. Application of the method to urine from normal persons and a patient with a brain tumor has been demonstrated.


Serological grouping of meningococci and encapsulated Haemophilus influenzae strains by latex agglutination. The latex agglutination method, utilizing antibody-coated latex particles, was adapted for serogrouping of Neisseria meningitidis and serotyping of encapsulated Haemophilus influenzae strains from agar plates. It was found to give more clear-cut results than conventional slide agglutination. A 100% agreement with the antiserum agar method was found for all strains isolated from blood or cerebrospinal fluid. Many meningococcal strains from nasopharyngeal carriers are autoagglutinable, but some of these gave a positive reaction with the group B latex reagent, although they were negative by the antiserum agar method. The latex agglutination method has several advantages over others: the lack of autoagglutination, easy performance, easy interpretation, and very low consumption of antisera.


Comparison of Minitek and conventional methods for the biochemical characterization of oral streptococci. Fifty-six strains of viridans streptococci were tested in the Minitek system (BBL Microbiology Systems, Cockeysville, Md.), and the results were compared with those obtained by conventional methods. An overall agreement of 98.9% was obtained when the Minitek tests were performed as follows. (i) All disks were incubated anaerobically for 48 h except for esculin and arginine, which required 5 to 7 days. (ii) The arginine disks were overlaid with 0.1 ml of sterile mineral oil even though incubated anaerobically. (iii) The Voges-Proskauer tests were performed under aerobic and anaerobic conditions. (iv) All tests for carbohydrate fermentation except for raffinose and salicin were read after the addition of 2 to 3 drops of 0.025% phenol red (pH 7.2). Of all the tests performed, only the fermentation of starch resulted in such poor agreement as to be considered unreliable. Results for inulin (unavailable in the Minitek system) were obtained by conventional methods.


Serodiagnosis of ram epididymitis by counterimmunoelectrophoresis, using Brucella ovis surface R antigen. The counterimmunoelectrophoresis (CIE) technique was developed as a diagnostic procedure for ram epididymitis caused by Brucella ovis. CIE test results with sera from naturally and experimentally infected male sheep compared favorably with those obtained by gel diffusion and complement fixation employing the same B. ovis surface R antigen. The main advantage of CIE over gel diffusion consists in a significant reduction of the time required to detect precipitin formation, whereas both methods obviate several of the difficulties encountered with complement fixation tests for B. ovis.


Identification of clinical isolates of mycobacteria with gas-liquid chromatography alone. Identification of 18 mycobacterial species was performed by analysis of profiles obtained by using gas-liquid chromatography. Organisms were saponified in methanolic NaOH, and the reaction mixture was treated with BF(3) in methanol and extracted with a hexane-chloroform mixture. An identification scheme was developed from 128 stock strains and tested against a collection of 79 clinical isolates. By using gas-liquid chromatographic profiles alone, 58% of specimens were correctly identified to species level, and an additional 41% were correctly identified to a group of two or three organisms. Use in a clinical laboratory over a 2-month period proved chromatography to be as accurate as and more rapid than concurrent biochemical testing. Of 81 isolates tested, 64% were identified to species level by chromatography alone. An additional 35% were differentiated to the same groups of two or three organisms as found in our analysis of stock strains. These groups consisted of: Mycobacterium tuberculosis, M. bovis, and M. xenopi; M. avium complex, M. gastri, and M. scrofulaceum; or M. fortuitum and M. chelonei. Identification to species level from these groups could usually be done by colonial morphology alone and could always be done by the addition of one selected biochemical test. This study demonstrated the practical application of gas-liquid chromatography in the identification of mycobacteria in a clinical laboratory. In particular, all strains of M. gordonae and M. kansasii were identified to species level. M. tuberculosis was definitively identified in 85% of cases. When it could not be definitely identified, the only alternatives were M. bovis and M. xenopi, both of which are rare causes of infection.


Isolation of acid-fast organisms from surgical specimens. A total of 204 surgical specimens positive for acid-fast organisms yielded 169 strains of Mycobacterium tuberculosis, 15 of which were isolated by Middlebrook 7H-9 broth medium only. Mycobacteria other than M. tuberculosis were isolated from 33 specimens, and Nocardia asteroides was isolated from 2 specimens.


Establishing and maintaining clinically healthy gingivae in Rhesus monkeys. The aim of the present investigation was to determine a method and a minimal frequency of plaque removal for the establishment and maintenance of clinically healthy gingivae in Rhesus monkeys. Six adult, male monkeys were used and divided into three groups. In each group different methods were used for plaque removal. Frequencies of plaque removal of one, two and three times per week for consecutive weeks were tested. The results showed that gingival health could be established and maintained with any of the methods used (toothbrushing and interdental flossing: toothbrushing and a 3-min application of 2% chlorhexidine gluconate) but only when plaque was removed 3 times a week. This frequency of plaque removal sessions is discussed in relation to the necessity for sedation of the animals with possible adverse side effects.


Magnetic susceptibility of microorganisms. Total magnetic susceptibility of 13 species and varieties of bacteria was investigated using the relative method of Guy. It has been established that the index of magnetic susceptibility is a constant characteristic of bacteria. Total magnetic susceptibility ranged from --0.3295.10(-6) in Escherichia P678 to --0.4965.10(-6) in Proteus. It has also been established that magnetic susceptibility changes during long-term passages of bacteria in fluctuating +/- 0.1 Oe) magnetic field. This is suggestive of a low threshold of their magnetic susceptibility and permits a rough assessment of the importance of fluctuations of the geomagnetic field for the viability of microbes.


Serum thyroid hormone concentrations and weight loss relationships in eight obese women during semistarvation. Eight obese female patients were studied over a period of 15 days whilst on 300 kcal diet. Serum levels of thyroxine and free throxine index were not altered significantly by semistarvation. A TRH test performed before and after the diet showed no appreciable change. Weight loss was intially rapid but later slowed despite good patients compliance. Serum concentrations of T 3 and reverse T 3 (rT3) early decreased (p less than 0.01) and increased (p less than 0.05) respectively, but returned towards control levels even before discontinuation of semistarvation. There was a positive correlation between the percentage decrease in body weight and the percentage increase in serum rT 3 (p less than 0.001), and a negative correlation between decrease in body weight and decrease in serum T 3 (p less than 0.001). Our results do not suggest that the variations in serum triiodothyronines limit the weight loss; it is probable, on the contrary, that the weight loss promotes the observed variations in thyroid hormones by as yet unknown adaptive metabolic forces.


The presence of antibody-coated anaerobic bacteria in asymptomatic bacteriuria during pregnancy. Quantitative anaerobic culture of urine samples obtained from 593 pregnant women by suprapubic bladder aspiration was performed to establish the involvement of anaerobic bacteria in asymptomatic urinary tract infections. The fluorescent antibody (FA) test was applied to the sediments of bladder aspirates to determine the site of infection. Anaerobic bacteriuria (greater than or equal to 10(4) microorganisms/ml of urine) was found in 34 patients, of whom five were FA-positive. These anaerobes were identified as Lactobacillus minutus, Veillonella parvula (two patients). Clostridium putrefaciens, and Peptostreptococcus anaerobius. Aerobic bacteriuria (greater than 10(4) microorganisms/ml of urine) was detected in 27 patients, of whom 13 were FA-positive. In 10 women with mixed aerobic/anaerobic bacteriuria, no FA-positive bacteria were found. The finding of FA-positive anaerobes may indicate that these organisms are involved in silent renal infection.


Kinetics of antibody production to group A and group C meningococcal polysaccharide vaccines administered during the first six years of life: prospects for routine immunization of infants and children. A cohort of infants, previously immunized once or twice between three and 12 months of age with vaccines containing polysaccharide of groups A and C Neisseria meningitidis, received boosters at two and five and one-half years of age with bivalent A/C vaccine. Antibody concentrations were measured by a radioactive antigen-binding assay. Concentrations of antibody to groups A and C N. meningitidis were 5.59 and 2.86 microgram/ml, respectively, by four years of age. After booster immunization at five and one-half years of age, concentrations of antibody to groups A and C N. meningitidis increased to 15.67 and 7.59 microgram/ml, respectively. Protective levels of antibody to group A meningococci may be achievable throughout early childhood by routine immunization with the A vaccine. Although the group C vaccine is effective in control of epidemics, the rapid decline in the concentration of antibody to group C meningococci following immunization of young children suggests that protection may not be long-lasting.


The role of natural IgG and complement in the phagocytosis of type 4 Neisseria gonorrhoeae by human polymorphonuclear leukocytes. The role of human serum components in the phagocytosis of logarithmic-phase type 4 Neisseria gonorrhoeae by human polymorphononuclear leukocytes was investigated. The requirement of fresh normal human serum (FHS) for optimal phagocytosis and the fixation of human immunoglobulin (IgG) and complenet (C3) to the gonococcal cell surface suggested that both serum factors participate in the phagocytosis of these organisms. The percentage of neutrophils containing ingested organisms was directly proportional to the concentration of IgG purified from FHS. Absorption studies suggested that this natural IgG binds to a trypsin-sensitive surface protein on type 4 gonococci and cross-reacts with stationary-phase type 2 N. gonorrhoeae, group C Neisseria meningitidis, and Branhamella catarrhalis, but not with logarithmic-phase type 2 gonococci or other Neisseria species. Although complement alone did not promote phagocytosis, it enhanced IgG-mediated ingestion. Studies using C2-deficient serum or serum chelators indicated that the alternative complement pathway participates in this interaction.


Mutagenicity studies with 5-thio-D-glucose. Cytogenetic investigation in mice, and the sex-linked recessive lethal gene test in Drosophila. Cytogenetic studies in bone marrow cells and dividing spermatocytes I in male mice and the sex-linked recessive lethal gene test in Drosophila melanogaster were initiated to evaluate the mutagenic potential of 5-thio-D-glucose, a reversible male contraceptive in some mammals. The compound was administered by oral intubation to mice at 40 or 60 mg/kg for various time periods. Oregon-R D. melanogaster males were fed medium containing 600 or 1200 ppm of the substance for 72 hours and mated with virgin Muller-5 females to test for gene mutation. The compound did not enhance structural and numerical chromosomal aberrations or elevate sex-linked recessive lethals in D. melanogaster. It is concluded that under the conditions of the present study 5-thio is a nonmutagenic male antifertility agent.


A solid-phase radioimmunoassay for the measurement of antibody to Aspergillus in invasive aspergillosis. A solid-phase radioimmunoassay (SPRIA) to measure antibody responses to Aspergillus fumigatus and Aspergillus flavus antigens in invasive aspergillosis (IA) was developed and compared with immunodiffusion (ID) and counterimmunoelectrophoresis (CIE). SPRIA detected significant elevations in levels of aspergillus antibody in 15 (79)% of 19 patients with IA. Fewer patients with IA were positive by ID (give of 19) or CIE (four of 19). Only seven )8%) of 58 subjects with other fungal or bacterial infections were positive by SPRIA, as was one (5%) of 20 by ID and CIE. Weak cross-reactivity between aspergillus and candida antigens was demonstrated by SPRIA. IgG levels in patients with IA and control subjects were equivalent. Thus, an antibody response to Aspergillus can be detected in a greater percentage of patients with IA by SPRIA than by ID or CIE. Although a few patients without aspergillosis had elevated levels of aspergillus antibody, a rise in antibody level was specific for IA.


Acquired immunity to human non-A, non-B hepatitis: cross-challenge of chimpanzees with three infectious human sera. Chimpanzees that had recovered from non-A, non-B hepatitis transmitted by inoculation of serum from each of three chronically infected humans were challenged by inoculation with a second of the three infectious sera to determine whether recovery from infection caused by one serum afforded protection against later infection by another. None of the challenge inoculations caused recognizable non-A, non-B hepatitis in any of the chimpanzees, a finding suggesting that either one agent or several agents sharing a common or similar antigen were responsible for the original non-A, non-B hepatitis in fections in these chimpanzees. Although circumstantial evidence in the literature suggests the existence of more than one agent of non-A, non-B hepatitis, the fact that the three inocula were obtained from humans residing in different geographic areas of the eastern United States suggests that one agent or a group of related agents may be the cause of many cases of non-A, non-B hepatitis in the United States.


Results of lithium- 131I treatment of thyrotoxicosis. The role of lithium as an adjunct to 131I therapy of thyrotoxicosis has been assessed. Seventeen lithium- 131I treated patients and 16 control- 131I treated patients have been followed for almost three years. Five lithium- 131I and 3 control- 131I patients have become hypothyroid with the lithium treated patients developing earlier thyroid failure. Ten lithium and 10 control patients remain clinically and biochemically euthyroid. Two lithium- 131I and 2 control- 131I patients remain on low dose antithyroid drugs.


Clinical responses of silvered leaf monkeys to infection with selected strains of Rickettsia tsutsugamushi. Minimal clinical and hematologic signs were observed in silvered leaf monkeys inoculated intradermally with four strains of Rickettsia tsutsugamushi, both virulent and avirulent for laboratory mice. The clinical response of the monkeys to the infection was related to neither the virulence of the strains in mice nor the antigenic characteristics of the strains.


Dynamics of correlated genetic systems. VI. Variation in recombination rates in experimental populations of Drosophila melanogaster. Recombination fractions among four chromosome 3 enzyme loci were measured over a 20 generation period in experimental populations of Drosophila melanogaster. The experimental populations were initiated with complete pairwise linkage disequilibria among the marker loci, thereby providing an initial structure favorable to selection for recombination modification. The recombination data show no evidence of directional changes in recombination rates among the marker loci, although variation in estimates, both between families within generations and between generations was large. These materials also yielded much lower estimates for recombination fractions for the centromeric region of the third chromosome than previously reported. Taken in total, the experimental results show no evidence for recombination modification, although the results do indicate ample genetic variance for recombination rates.


Bacteriology of rattlesnake venom and implications for therapy. Although the incidence of infection secondary to the bites of venomous snakes remains unknown, the routine use of prophylactic antimicrobial therapy is advocated. In this study, the venom from 15 rattlesnakes was cultured, and 58 aerobic and 28 anaerobic strains of bacteria were isolated. The most common species isolated were Pseudomonas aeruginosa, Proteus species, coagulase-negative staphylocci, and Clostridium species. Bacteroides fragilis was also recovered. When the fang sheaths of four additional rattlesnakes were retracted and the fangs of these snakes decontaminated, 50% of the samples of venom had no bacterial growth (P = 0.035). Until a clinical study is performed, the use of antimicrobial therapy that reflects that complex oral flora of rattlesnakes is still recommended in most cases of envenomization.


Inhibitory effect of heat-labile serum factors on detection of staphylococcal, pseudomonas, and hepatitis B surface antigens by solid-phase radioimmunoassay. Specific antibodies have previously been found to impair detection of staphylococcal antigen by solid-phase radioimmunoassay. The inhibitory effect of heat-labile serum factors on the detection of antigen is now described. Heating of serum at 56 C for 30 min prior to addition of antigens of Pseudomonas aeruginosa or Staphylococcus aureus improved sensitivity by eight- to 32-fold. Heating was less effective when performed after addition of antigen. Treatment with zymosan reduced the inhibitory effect of serum, although less effectively than did preheating. Antigen in buffer was detected with 16 times less sensitivity in antibody-coated tubes exposed to fresh serum than in tubes exposed to heated serum. These findings suggest that complement factors can interact with antibody in coated tubes and at least some antigens and thereby inhibit detection of antigen. The finding that heat treatment improved the sensitivity of Ausria II -125 (Abbott Laboratories, North Chicago, Illinois) for hepatitis B surface antigen is of potential immediate clinical applicability.


[Immunologic studies concerned with high dose radiotherapy for osteosarcoma (author's transl)]. The 5-year cumulative survival rate was measured in 28 cases of osteosarcoma treated with high dose radiation since 1969 is 48.8% in our clinic. It can be said that high dose radiotherapy has a significant survival effect compared to early amputation therapy for the patient with osteosarcoma. The difference of the prognosis between both therapies may be related to immunological reactions. In order to obtain further information on this possibility, experimental studies on mice suffering from tumors have been performed. Results revealed that spleen cell migration inhibition reaction, as a specific immunity, became negative and anti-tumor properties were eliminated as a results of the amputation of the limb bearing the tumor. Also, when BCG as well as irradiated tumor cells were administered to tumor-afflicted mice, an improved rate of survival among the mice was observed. As a result of the study of patients with osteosarcoma that has been treated with high dose radiation related to changes in their immunity, it was disclosed that there was a marked tendency to diminution in peripheral blood lymphocytes or T cells in cases with poorer prognoses. In cases of long survival, both showed high values. Lymphoblastgenesis by PHA and PWM showed higher values in cases with better prognoses than in those with poor prognoses. Furthermore, in many of the cases in which the tuberculin skin reaction became negative, a short survival period was noted.


Acquisition of prerequisites to conservation by macaques. Through a progressive series of learning sets, two rhesus macaques were trained to discriminate transformations that produce change of substance from other transformations and manipulations. Several tests were administered to assess the generalization of this discrimination to a variety of objects and shapes and to assess the effects, if any, of size cues, experimenter familiarity, and various methods to problem presentation. After training was completed, performance was not greatly affected by differences in problems and presentations. The results of these experiments suggest that monkeys can acquire this prerequisite for substance conservation as a result of learning set training.


Stimulus similarity and order as factors in visual short-term memory in nonhuman primates. The short-term retention of nonhuman primates for a single sample or for two successively presented samples was assessed in four delayed matching-to-sample experiments with delays of .03, 4, 8, 16, and 32 sec. The single sample tasks included one (Experiment 1) or two (Experiment 4) distractor stimuli in the choice set (matching test). In the two successive samples tasks, the animals matched (reconstructed) the order of presentation of two samples with (Experiment 3) and without (Experiment 2) a distractor stimulus. Also, the possible combinations of eight stimuli (four colors and four shapes) were arranged to test the effects of sample set and choice set similarity. Taken together, analyses of the errors indicated that both sample and choice set similarity were significant sources of confusions in delayed matching. Order errors occurred independently of similarity but were a source of forgetting primarily at the longest delays (16 and 32 sec). Two exceptions to the similarity effect (second response errors in Experiment 3 and errors of an inexperienced group in Experiment 4) were observed. Possible reasons for the exceptions and several implications of these findings for theories of short-term memory are discussed.


Effects of lysozyme on Bacillus cereus 569: rupture of chains of bacteria and enhancement of sensitivity to autolysins. Bacillus cereus 569 is known to be resistant to lysis by lysozyme because of the presence of deacetylated glucosamine residues in its peptidoglycan, and cultures continued to grow even in the presence of lysozyme at 200 microgram ml-1. However, lysozyme caused rupture of the chains of bacteria and promoted the rate of autolysis in a non-growing cell suspension, causing a doubling of the rate of release of radioactively labelled wall material. Heat-inactivated cells did not autolyse and were not lysed by lysozyme unless they were supplemented by unheated cells or cell-free autolysate. Enhancement of autolysin activity could also be effected by pre-treatment of heated cells with lysozyme. The action of lysozyme on isolated cell walls released some free reducing groups, indicating limited breakage of the polysaccharide chains of peptidoglycan, and it was concluded that lysozyme modified the peptidoglycan and made it more susceptible to autolysin(s). Lysozyme also enhanced the rate of septum separation and the probable significance of the results in relation to the control of cell separation is discussed.


Asbestos fibers: 1. A modified preparation of tissue samples for analysis by electron microscopy. 2. Presence of fibers in tissues of baboon fed chrysotile asbestos. A specimen preparation technique is described which yields uniformly dispersed particles on transmission electron microscope (TEM) grids. It is relatively rapid and probably incurs no fiber loss. This technique was used to test the hypothesis that orally administered chrysotile asbestos may pass through the wall of the gastrointestinal tract (GI) of the neonate baboon. A test neonate baboon was bottle-fed chrysotile asbestos which was suspended in a milk formula. Control and test baboons were sacrificed at the end of a 9-day feeding period, and several tissues were removed and preserved. Kidney, liver, and spleen tissues were prepared for TEM analysis. Tissues from the control animal showed no fibers. The number of chrysotile fibers recovered from the test kidney cortex was significantly higher than the level found in the control kidney cortex (p less than 0.005). The frequency distribution of fibers lengths in the kidney cortex was very similar to that of the feeding suspension. This study suggests that a wide range of fiber sizes can penetrate the GI tract and migrate.


Differing contribution of polymorphonuclear cells and macrophages to protection of mice against Listeria monocytogenes and Pseudomonas aeruginosa. Bacterial growth and lethality of Listeria monocytogenes in mice were augmented by carrageenan-treatment and X-irradiation (8 J kg-1), whereas growth and lethality of Pseudomonas aeruginosa were augmented by X-irradiation but not by carrageenan-treatment. Protection against L. monocytogenes, at least in the early phases, appears to depend mainly on macrophages, since carrageenan depletes macrophages but not polymorphonuclear cells (PMN), whereas protection against P. aeruginosa appears to depend mainly on PMN. Ineffectiveness of PMN in elimination of L. monocytogenes is supported by histological examination and observation of intracellular killing in vitro.


Chemotaxis towards sugars by Bacillus subtilis. Many sugars and derivatives were tested in the capillary assay for their attraction of Bacillus subtilis. The major attractants were 2-deoxy-D-glucose, D-fructose, gentiobiose, D-glucose, maltose, D-mannitol, D-mannose, N-acetylglucosamine, alpha-methyl-D-glucoside, beta-methyl-D-glucoside, N-acetylmannosamine, alpha-methyl-D-mannoside, D-sorbitol, L-sorbose, sucrose, trehalose and D-xylose. Only glucose chemotaxis was completely constitutive. Competition experiments were carried out to determine the specificities of chemoreceptors. There were 25 instances of no influence of two sugars on each other's taxis, 92 instances of one sugar interfering non-reciprocally with chemotaxis towards another and 49 instances of two sugars reciprocally competing. However, in most of the last instances, other sugars were identified that interfered with chemotaxis towards one member of the pair but not the other. Thus, nearly all sugars and related compounds appear to be detected by their own chemoreceptors, but many secondary interactions exist.


Partial purine deprivation causes sporulation of Bacillus subtilis in the presence of excess ammonia, glucose and phosphate. In strains of Bacillus subtilis able to synthesize purines de novo, massive sporulation is suppressed by the combination of excess ammonia, glucose and phosphate. Purine auxotrophs, blocked in the general or the guanine-specific portion of the branched purine pathway, sporulated in such a medium when the purine required for normal growth was removed from the medium. The resulting spore titre and the sporulation frequency increased with the residual growth rate in the purine-free medium, i.e. with the leakiness of the purine mutation. Sporulation was further increased by allowing residual growth in growth-limiting amounts of guanosine. None-leaky purine mutants blocked before 5'-phosphoribosyl-5-amino-4-imidazole carboxamide also sporulated well when supplied with 5-amino-4-imidazole carboxamide at concentrations (2 mM) that supported growth at a suboptimal rate.


Lysine regulation of penicillin biosynthesis in low-producing and industrial strains of Penicillium chrysogenum. The inhibitory effect of L-lysine on penicillin biosynthesis by Penicillium chrysogenum has been compared in a low-producing strain (Wis. 54-1255) and a high-producing strain (ASP-78). Lysine inhibited total penicillin synthesis to a similar extent in both strains. However, in the high-producing strain the onset of penicillin synthesis occurred even at a high lysine concentration, whereas in the low-producing strain lysine had to be depleted before penicillin production commenced.


Lipoquinones of some spore-forming rods, lactic-acid bacteria and actinomycetes. The respiratory quinones of 73 strains of Gram-positive bacteria including spore-forming rods, lactic-acid bacteria and actinomyctes were examined. Menaquinones with seven isoprenoid units (MK-7) were the main quinone type found in representatives of the genus Bacillus and in Sporolactobacillus inulinus. However, a strain of B. thuringiensis produced MK-8 in addition to MK-7, and strains of B. lentus and B. pantothenticus appeared to produce MK-9 and MK-8, respectively, with no MK-7. In the clostridia and lactic-acid bacteria, no quinones were found, except in Pediococcus cerevisiae NCTC 8066 and Lactobacillus casei subsp. rhamnosus ATCC 7469, which contained menaquinones, and Streptococcus faecalis NCTC 775 and HIM 478-1, which contained demethylmenaquinones, in relatively low concentrations. Menaquinones were also found in the actinomycetes (except Actinomyces odontolyticus and Bifidobacterium bifidum which did not produce any quinones) and in Protaminobacter alboflavus ATCC 8458, the so-called Actinobacillus actinoides ATCC 15900 and Noguchia granulosis NCTC 10559.


Preparation of protoplasts and whole cell ghosts from Mycobacterium smegmatis. Cell wall-deficient forms of Mycobacterium smegmatis were produced in growth medium containing D-cycloserine and horse serum. These cells were transformed into protoplasts with EDTA and lysozyme. Subsequent lysis by nucleases followed by osmotic shock produced membrane vesicles (whole cell ghosts).


Biochemical and antigenic comparison of the envelope glycoproteins of Venezuelan equine encephalomyelitis virus strains. Pulse-chase experiments after synchronous initiation of translation indicate that the larger Venzuelan equine encephalomyelitis (VEE) virus membrane glycoprotein E2, is derived by proteolytic cleavage of the precursor, PE2. The structural proteins of VEE virus strains representing each of the antigenic subtypes and varieties have been compared by discontinuous SDS-polyacrylamide gel electrophoresis. Nucleocapsid proteins of all isolates were similar in size (mol. wt. 35 to 36 X 10(3). The mol. wt. of E1 varied from 48 to 51 X 10(3) and the mol. wt. of E2 glycoproteins ranged from 53 to 59 X 10(3). Pixuna virus contained a third envelope glycoprotein of 59 X 10(3) mol. wt. in addition to the two major glycoproteins of mol. wt. 53 X 10(3) and 48 X 10(3) respectively. The isoelectric points (pI) of E1 and E2 for all VEE strains studied were approx. 7 and 9 respectively. Both glycoproteins of TC-83 virus induced precipitating antibodies which reacted only with the homologous purified E1 and E2 glycoproteins. Antibodies to E2 protein of each virus neutralized virus infectivity and inhibited the agglutination of goose erythrocytes by virions. Haemagglutination-inhibition tests using antisera to E2 glycoproteins of prototype viruses, representing each of the antigenic subtypes and varieties, differentiated the viruses into subtypes I, II, III and IV with subtype I divided into variants 1AB, 1C, 1D and 1E.


An enzyme-linked immunosorbent assay (ELISA) for PBS Z1, a defective phage of Bacillus subtilis. The sensitivity, reproducibility and specificity of an enzyme-linked immunosorbent assay (ELISA) for the defective phage PBS Z1 of Bacillus subtilis have been investigated. It was shown that phages in concentrations between 10(8) and 2.5 X 10(10) particles/ml could be assayed with this method. The coefficient of variation for concentrations between 5 X 10(8) and 5 X 10(9) particles/ml was approx. 10%. From some other Bacillus phages tested, only the defective phages resembling PBS Z1 in morphology were detected efficiently with the ELISA for PBS Z1. A comparison is made between ELISA and other assays for PBS Z1.


Serologic response in human hepatitis A: detection of antibody by radioimmunoassay and immune adherence hemagglutination. An indirect solid-phase radioimmunoassay (RIA) for detection of antibody to the hepatitis A antigen (anti-HAV) was developed using polystyrene pearls as the solid phase and hepatitis A antigen (HAAg) extracted from marmoset livers. This RIA was compared to an immune adherence hemagglutination assay (IAHA) which employed HAAg derived from the stools of chimpanzees collected during acute hepatitis A. Anti-HAV was detected in the sera of 15 humans with naturally acquired hepatitis A infection. Sensitivity and specificity were greater using the RIA, permitting the detection of anti-HAV as early as the time of onset of jaundice. Either seroconversion or a significant increase in the titer of anti-HAV was demonstrated following hepatitis A exposure in paired sera from six patients by both techniques. No significant difference in anti-HAV responses was noted between patients with icteric compared to anicteric hepatitis A or between children and adults with hepatitis A.


Adenine arabinoside inhibition of adenovirus replication enhanced by an adenosine deaminase inhibitor. The inhibition of adenovirus multiplication by adenine arabinoside was determined by yield reduction in one-step multiplication cycle. Inhibition was greatly enhanced by an adenosine deaminase inhibitor (2-deoxycoformycin) in concentrations down to 10 ng/ml. Adenovirus types from four subgroups showed similar results. However, the enhancing effect of adenosine deaminase inhibitor was great in HeLa cells, moderate in human fibroblasts, and negligible in Vero cells. This difference could be explained by different concentrations of adenosine deaminase found in cell homogenates.


The occurrence of mononuclear cells at sites of osteoclastic bone resorption in experimental periodontitis. The placement of silk ligatures around the necks of teeth and into the gingival sulcus causes a rapid, acute inflammatory response leading to vigorous osteoclastic resorption of alveolar crestal bone. Associated with the large numbers of osteoclasts are mononuclear cells, predominantly fibroblast-like cells and macrophages. Some fibroblast-like cells contain intracellular collagen fibrils. It is suggested that in periodontal disease these mononuclear cells may compliment the action of osteoclasts by ingesting and degrading matrix molecules mobilized from bone but not ingested or degraded by osteoclasts.


Early post traumatic seizures in adults. Study of 84 cases. In a series of 2.980 brain-injured patients, 84 (2.8%) showed epileptic seizures. Epilepsy occurred with a higher incidence rate in operated patients (11.6%) than in non operated ones (1.2%). Seizures appeared within the first week since trauma in 91% of the cases. They have been single in 48%, recurring in 34%, with figures of status epilepticus in 16% partial in 61%, and GM in 33%. Prognosis quoad vitam of early epilepsy seems to be favourable, however, in severely brain-injured patients, seizures occur more frequently and may determine a worsening of the clinical picture even if they are not responsible for the death.


Effect of adenosine triphosphate and some derivatives on cerebral blood flow and metabolism. 1. Responses of cerebral blood vessels to peri- and intravascular doses of ATP (adenosine triphosphate) and some derivatives were studied in cat and baboon. 2. Perivascular application of ATP to cat pial arterioles gave a threshold dilatory effect at a concentration of 10(-11) M. This figure is comparable to the amount of ATP calculated to be released from electrically stimulated brain slices. 3. It is concluded that adenine nucleotides have a major role to play in the local control of cerebral blood flow. 4. Intracarotid injection of ATP showed a calculated threshold effect at 4 x 10(8) M in the cat and 4 x 10(-9) M in the baboon. 5. The threshold response of the vasculature to intracarotid adenosine lay between 4 x 10(-7) M and 4 x 10(-6) M in the baboon. Little effect was produced with AMP, pyrophosphate and inorganic phosphate. 6. Intracarotid ATP increased the oxygen consumption of the baboon brain parenchyma. This effect was attributed in part to an elevation of the cellular cyclic AMP levels. 7. Osmotic disruption of the blood-brain barrier in baboon did not affect the vasodilatory or metabolic effect of intracarotid ATP. 8. It is postulated that circulating purine compounds mediate a form of metabolic communication inthe body. Also, release of purine compounds from active local nerves might influence cerebral blood flow.


[Double contrast studies of the colon. Use of acetyl-cystein or mucofluid? (author's transl)]. The authors propose a replacement product for acetylcysteine which had been suggested for use in order to obtain good double contrast studies of the colon after retrograde barium introduction. Mucofluid gives even better results at a much lower cost.


Intrauterine infections in nonhuman primates. Gross and histologic examination of five nonhuman primate placentas revealed inflammatory processes, either of the ascending type, with chorioamnionitis and fetal vasculitis, or of the hematogenous type with villitis. These reactions were similar to those occurring in man, with known implications for perinatal outcome.


Amniocentesis and antenatal sex determination in the rhesus monkey (Macaca mulatta). Antenatal sex was correctly identified in 87.5% of the rhesus monkeys studied by evaluation of sex chromatin bodies in the exfoliated cells present in amniotic fluid. Fluid samples were obtained by either amniocentesis, amnioscopy, or by cesarean section at 80--120 days of gestation. Amniotic fluid concentrations of progesterone, total estrogen, and testosterone were correlated with fetal sex. Concentrations of these hormones decreased over the time period examined. Amniocentesis is a simple procedure which provides for routine fetal sex determination.


Menetrier's disease in a rhesus monkey (Macaca mulatta). Gross and microscopic features closely resembling those found in Menetrier's disease in man are described in a 20-month-old rhesus monkey. The gastric lining was characterized by greatly enlarged rugae caused by mucosal hypertrophy and hyperplasia along with outfolding of the muscularis mucosa and the submucosa. The mucosa and submucosa were infiltrated with inflammatory cells, mainly lymphocytes and plasma cells.


Alteration in the distribution of organ perfusion following profound hypothermia and circulatory arrest in the baboon. The physiologic consequences of profound hypothermic circulatory arrest in infants are incompletely understood. Immature baboons underwent surface cooling, followed by core cooling using cardiopulmonary bypass, circulatory arrest for 30 min, and perfusion rewarming. Blood flow to and within organs was studied using the multiple-radionuclide-label microsphere technique. Marked redistribution of total and regional myocardial and cerebral flow occurred during cooling and rewarming.


Altered lymphocyte response in adjuvant-induced arthritis. Evaluation of lymphocyte response has been undertaken in adjuvant-induced arthritis in rats. Lymphocyte reactivity was studied by evaluating lymphocyte transformation in mixed lymphocyte-chondrocyte cultures as well as the response to PHA and M. tuberculosis. It was found that animals immunized with Freund's complete adjuvant showed increased transformation when challenged with M. tuberculosis powder. However, there was a depressed response to PHA and a markedly depressed response in mixed lymphocyte-chondrocyte cultures.


Persistence of impaired reversal learning in young monkeys exposed to low levels of dietary lead. Lead acetate in milk was fed daily to infant rhesus monkeys at doses averaging 0 (control), 0.287 (low-Pb), or 0.880 (high-Pb) mg/kgd for the first year of life. Pb concentrations in whole blood (PbB) averaged 4.15, 31.71, and 65.17 microgram/dl for the control, low-Pb, and high-Pb groups, respectively, during the year of treatment and declined toward control levels when Pb dosing was stopped. Behavioral observations during the year of treatment had shown that both experimental groups were retarded in their acquisition of object-cue discrimination reversal learning sets. At 4 yr of age, when PbB levels in all animals were normal, the ability of the same monkeys to acquire a series of 3 spatial-cue reversal learning sets was examined; these data form the basis for this report. In the first problem, the high-Pb group was significantly retarded in acquisition of the original discrimination and of most reversals, and the low-Pb group was retarded on reversal 1 only. These deficits declined in severity across the three problems administered, in a manner similar to that seen in the tests given during the first year of life. These data demonstrate that reversal learning retardation, observed early in life, can recur in postadolescent primates with a history of chronic, low-level Pb intoxication during infancy.


Mammary tumorigenesis in the rat following prenatal exposure to diethylstilbestrol and postnatal treatment with 7,12-dimethylbenz[a]anthracene. Pregnant rats were injected with vehicle or 1,2 microgram diethylstilbestrol (DES) during wk 2 or 3 of gestation; their female offspring ( approximately 50 d old) were fed 7,12-dimethylbenz[a]anthrocene (DMBA). The survivors (27 per group) were sacrificed 30 wk later. The three groups did not differ in the number of tumor-bearing animals; however, significantly more palpable mammary tumors arose in both DES-exposed groups than in controls. When DES was given during the second trimester, palpable tumors appeared earlier than in the other two groups. Thus, transplancental exposure to DES potentiated the action of a known carcinogen (DMBA) on rat mammary tissue. These results raise the possibility that, for young women, DES exposure in utero may have affected tissues other than the vagina. Further investigation is warranted, with special emphasis on the effects of DES on mammary and other estrogen-sensitive tissues.


Alpha-chain disease--a report of eleven patients from Iran. About one hundred cases of alpha-chain disease have been reported to date from various Mediterranean and Middle-Eastern countries. We report eleven Iranian patients with primary lymphoma of the duodenum and upper small intestine and positive alpha-chain protein in their sera. The clinical, laboratory and pathological findings in these patients are discussed. The initial favourable response of some of these patients to antibiotic treatment is noted. It is concluded that the optimal treatment of alpha-chain disease is not yet well established, and the clinical improvement might not coincide with histological and immunological remission.


The management of penicillin resistant gonococcal urethritis in Northern Nigeria. Forty-five men, with acute gonococcal urethritis, were treated with a single dose regime of Probenecid 1 gm and Procaine penicillin, 4.8 mega-units, yielding a cure in 97 per cent. Multiple antibiotic resistance was noted in 69 per cent of 28 Neisseria gonorrhoeae isolates, and of 19 isolates tested for precise penicillin sensitivity, 58 per cent were resistant to greater than 2 i.u./ml. Serum penicillin levels measured in 3 patients showed adequate duration of plateau levels. The advantages of adopting such a regime are discussed.


Serum enzyme, bilirubin, uric acid, and cholesterol concentration changes during 7 days of total parenteral nutrition. Lactic dehydrogenase, glutamic oxalacetic transaminase, and alkaline phosphatase remained within normal limits during 7 days of total parenteral nutrition. Five patients showed a rise of total bilirubin up to 1.1 mg/100 ml on the fifth postoperative day, with no change in serum enzymes. Uric acid concentration fell continuously up to the third postoperative day (but within normal limits) and remained decreased until the end of infusion. During the preoperative nutrition period and after surgical intervention, cholesterol concentration showed a steep fall. Very low cholesterol concentrations can cause an alteration of erythrocyte membrane synthesis and should, therefore, be avoided.


Intravenous protein-sparing therapy in patients with gastrointestinal disease. Two groups (each of 6 moderately ill, protein-depleted patients) were infused daily for 7 days. Mean 7 day nitrogen (N) balances with infusions of 0.83 and 1.83 g of a defined amino acid mixture (containing further nutrients but no other source of energy)/kg ideal body wt/day were -3.66 and +1.54 g/day, respectively (P less than 0.025) when adjusted for changes in body urea and estimated miscellaneous N losses. Concentrations of plasma free fatty acids, immunoreactive insulin and glucagon, and of blood glucose, pyruvate, lactate and glycerol were indistinguishable on corresponding treatment days in the 2 groups but blood ketone bodies were lower in the 1.83 g/kg group. Blood amino acid concentrations of alanine, valine, leucine, and isoleucine were similar, whereas those of phenylalanine, histidine, serine, and arginine were higher, and glutamine lower, in the 1.83 g/kg group. The data confirm that not only can body protein mass be maintained, but a net positive N retention achieved, in such patients, through provision of exogenous amino acids and concurrent mobilization of endogenous energy stores. Of note is that this fat mobilization can occur without plasma free fatty acids and/or significant blood ketone body elevations. An infusion of 2, rather than 1 g/kg/day seems suitable in the situation examined.