Cellular immunity to bacteria: impairment of in vitro lymphocyte responses to Pseudomonas aeruginosa in cystic fibrosis patients. Lymphocyte responses to the mitogens phytohemagglutinin and concanavalin A and to Streptococcus pyogenes, Staphylococcus aureus, Haemophilus influenzae, and Pseudomonas aeruginosa were evaluated in patients with cystic fibrosis and in normal individuals. Lymphocyte proliferation in vitro was stimulated by gentamicin-killed whole bacteria, and the proliferative response was measured by [3H]thymidine incorporation. The in vitro lymphocyte responses to antibiotic-killed bacterial reached maximum thymidine incorporation after 5 days in culture and followed a unimodal dose-response curve for each of the bacteria studied. A significant specific incapacity to respond to P. aeruginosa was detected in cystic fibrosis patients with advanced clinical disease.


Development of antitoxin with each of two complementary fragments of Clostridium botulinum type B derivative toxin. Two fragments with molecular weights of 111,000 (fragment I) and 59,000 (fragment II) were separated from each other by gel filtration of dithiothreitol and urea-treated, trypsinized derivative toxin (molecular weight, 170,000) of the proteolytic Okra strain of Clostridium botulinum type B on a column of Sephadex G-200 (superfine) with a buffer containing dithiothreitol and urea. Upon removal of dithiothreitol and urea by dialysis, the two fragments reassembled to reconstruct the derivative toxin molecule. Both fragments were immunogenic, and both anti-fragments neutralized type B toxin. The neutralizing activities of both anti-fragment I and anti-fragment II were, however, lower than that of the anti-derivative toxin, suggesting that the molecular integrity of derivative toxin is essential for sufficient production of the neutralizing antibody. The immunological difference found between type B toxin from a proteolytic strain and that from a nonproteolytic strain was ascribed to the antigenic difference of fragment I.


Species specificity of attachment and damage to oviduct mucosa by Neisseria gonorrhoeae. Neisseria gonorrhoeae colony type 1 were inoculated into organ cultures of oviducts obtained from various animal species. Gonococci rapidly attached to extensive areas of the mucosa of human oviducts (fallopian tubes), entered the mucous-secreting cells, and caused histological damage to the tissues. In addition, 2 to 4 days after infection there was complete loss of ciliary activity. In contrast, gonococci attached very scantily or not at all to the mucosa of rabbit, porcine, or bovine oviducts. However, the organisms multiplied in the medium of these organ cultures and were located sometimes in the base of mucosal cells and in large numbers in the submucosa. Despit this, there was no histological evidence of damage, and at least 7 days after infection ciliary activity was maintained equally as well as it was in uninfected control cultures. The host specificity of N. gonorrhoeae appears to be determined, at least in part, by a markedly diminished ability of the organisms to attach to and damage the genital mucosa of nonhuman species.


In vitro guinea pig leukocyte reactions to Rickettsia rickettsii. The presence of cell-mediated immunity in Rocky Mountain spotted fever-infected guinea pigs was determined by two in vitro assays: whole blood lymphocyte transformation (LT) and macrophage migration inhibition. Increased LT was detected as early as 1 week in guinea pigs infected with Rickettsia rickettsii and treated with oxytetracycline and was detected by two weeks in infected but untreated guinea pigs. Elevated LT was still detectable at 10 weeks postinfection. Guinea pigs vaccinated with killed rickettsiae failed to develop lymphocyte responsiveness; however, there was a rapid lymphocyte response after challenge with live organisms, suggesting potentiation by the vaccine. Vaccinated guinea pigs that were challenged and then treated with antibiotic failed to develop LT, suggesting that infection is necessary for the observed response. Macrophage migration inhibition was detected in both infected and vaccinated guinea pigs by 1 week after infection, but this response was no longer detected 4 to 5 weeks later. Antibody appeared at 2 to 3 weeks postinfection and was present at low levels through week 10. Antibody-treated rickettsiae were phagocytized and destroyed by guinea pig peritoneal macrophages, whereas normal serum-treated rickettsiae replicated and eventually destroyed the phagocytes.


Biologic assessment of copper-containing amalgams. In order to reduce creep and avoid marginal fractures in amalgam restorations, new alloys containing higher proportions of copper have been introduced. Fillings of these materials were placed in cavities prepared in the deciduous teeth of monkeys or placed in polyethylene tubes and implanted subcutaneously in rats. Conventional silver/tin alloys and zinc oxide eugenol cement were used as reference materials. Despite limitations due to the varying depths of cavities and the small number of animals involved it was concluded that the high copper alloys caused more severe pulp damage than the other materials studied. In the implantation studies many of the high copper specimens were exfoliated before the end of the experimental period. It is concluded that in deep cavities these materials require the use of a non-toxic base or lining material although as they are commonly used in young children's teeth the placement of linings and the isolation of the cavity pose problems.


The difference in effect of predosing with antiallergic compounds between the rat PCA model and the monkey asthma model. Comparison of the effects of PRD-92-Ea and disodium cromoglycate. DSCG and PRD-92-Ea showed tachyphylaxis in the rat PCA test when a large intravenous dose (40 mg/kg of DSCG and 20 mg/kg of PRD-92-Ea) was given 30 min before administering intravenously the expected effective doses at the time of antigen challenge. This predosing led to a marked loss of effectiveness. Cross-reacting tachyphylaxis was also demonstrated in this model. Predosing rats with DSCG led to a great loss of effectiveness of a subsequent expected active dose of PRD-92-Ea and predosing with PRD-92-Ea had the same effect in preventing the antiallergic action of a subsequent dose of DSCG. The Rhesus monkey (Macaca mulatta) did not exhibit this phenomenon of tachyphylaxis. PRD-92-Ea and DSCG were given intravenously 30 min before and the same dose of 20 mg/kg repeated just before Ascaris suum challenge in the Rhesus monkey asthma model with no loss of their expected effectiveness.


Studies on corynebacterial precipitinogens common to Mycobacteria, Nocardiae and rhodochrous. Ten strains of Corynebacterium were analyzed by means of the comparative immunodiffusion technique employing reference precipitation systems from strains of Mycobacterium, Nocardia and the rhodochrous taxon. The test strains had precipitinogens in common with the reference strains. Two of the intergenerical cross-reacting precipitinogens revealed were labelled x and y. Two other precipitinogens (alpha,beta), earlier found to be common for strains of mycobacteria, nocardiae, and rhodochrous, were not demonstrated in the corynebacterial strains, thus indicating a qualitative or quantitative serological difference between Corynebacterium and the other taxa. It was furthermore shown that the corynebacterial preparations reacted with antisera against ribosomal 30S fractions from mycobacteria. Common precipitinogens in ribosomal preparations from mycobacteria, nocardiae, rhodochrous, and corynebacteria are discussed.


A precipitate adsorption on surface technique: a combination of immunodiffusion and thin-layer immunoassay. A new technique--precipitate adsorption on surface--is described. The technique implies a combination of immunoprecipitation in gel and diffusion-in-gel-thin-layer immunoassay. The principles of the technique are illustrated by a series of model experiments.


Rh antibodies produced by an isoimmunized chimpanzee; reciprocal relationship between chimpanzee simian-type isoimmune sera and human anti-Rho reagents. An isoimmune chimpanzee serum was found to contain, in addition to expected antibodies of the simian-type specificity anti-Lc, a fraction of anti-Rho specificity, as proved by parallel tests with standard human anti-Rho sera as well as by absorption experiments using human and chimpanzee red cells. On the other hand, the two human anti-Rho sera tested in parallel were found to contain not only antibodies of specificity anti-Rho but also antibodies capable of detecting Lc specificity on the chimpanzee red cells. The reciprocal relationship between human and chimpanzee isoimmune sera constitutes the first example of symmetrical cross-reactivity between closely related species. The new source of anti-Rho reagents is expected to contribute additional information on the human Rh-Hr blood group system.


Scintigraphy in diagnosis of osteomyelitis of the jaws. The symptoms of an acute osteomyelitis of the jaws are often uncharacteristic, and typical radiographic changes usually do not appear until after the first weeks of disease. Even when such changes are established it is difficult to distinguish an active infectious disease from lasting changes due to a sterilized osteomyelitis. Scintigraphy with bone-seeking radiopharmaceuticals appears to be a valuable diagnostic technique. A case of osteomyelitis of the mandible is reported.


Computer utilization in an oral biopsy service. In order to improve our knowledge about the biological activity of oral lesions, a computer-based system of oral biopsy data recall has been established. The program is utilized in eight United States dental schools and is adaptable to almost any oral biopsy service with an available computer. This article deals with the implementation of the program at Washington University School of Dental Medicine.


Postoperative jaw pain--floctafenine versus dihydrocodeine. A double-blind single dose trial was made of floctafenine 200 mg versus dihydrocodeine 30 mg in 100 patients complaining of jaw pain following oral surgery under general anaesthesia. There was no statistical difference between the two analgesics for the whole group, but dihydrocodeine was better for the relief of more severe pain.


Extraosseous calcifying epithelial odontogenic tumor (Pindborg tumor). A 20-year-old man presented with a painless enlargement of the maxillary gingiva with no bone involvement. Microscopic examination with special stains confirmed the diagnosis of a clear cell variant of a calcifying epithelial odontogenic tumor (Pindborg tumor). This is the second reported case of such a variant in an extraosseous location.


Scleroma (rhinoscleroma) in a Nigerian maxillo-facial practice. Review and case reports. Scleroma (rhinoscleroma), a chronic granulomatous disease of the upper respiratory tract, is endemic within a confined distribution of geographic foci. Sporadic cases and those arising in poorly defined foci may be unexpected and difficult to diagnose. Three cases from a maxillo-facial unit in northern Nigeria are described, of which two simulated intranasal neoplasms and the third had atypical clinical features. These cases form the first substantial description of scleroma in Nigeria, a country that is part of an indefinite focus of the disease. The epidemiology, clinical features, pathology and diagnosis of scleroma are reviewed.


Pilomatrixoma in the differential diagnosis of pathologic calcification of the paraoral soft tissue. As an incidental finding a firm nodule was palpated in the skin overlying the right angle of the mandible in a 78-year-old female. The radiograph showed a dense calcification. Histologically the lesion proved to be a pilomatrixoma. A brief review of the literature of the pilomatrixoma is presented. Finally, some comments are made about calcification in the paraoral region, especially in the lateral aspect of the neck.


A simplified model for teaching minor oral surgery. A teaching model, specially constructed for the purpose of training senior dental students in the use of surgical instruments, is described. The advantages of this model are inexpensiveness and reproducible simulated surgical situations securing a uniform teaching objective.


The prophylactic use of Thio-TEPA, an adjuvant in the treatment of bladder tumours. In order to reduce the recurrence rate of bladder tumours, 78 patients with bladder tumours have been treated since November 1973 with topical Thio-TEPA bladder instillations. This method was used as an adjuvant to other forms of therapy. At the time of this review 20 of the patients have been followed up for at least one year following the onset of Thio-TEPA treatment. The total number of recurrences in these patients seems to be reduced. Advantages of topical therapy and indications for treatment are discussed.


Isolation of transitional epithelial cells from the rat urinary bladder. A method was developed to isolate transitional cells from the mucous lining of the rat urinary bladder by treatment with ethylene diaminetetraacetic acid and sonication. The isolated cells were examined by phase contrast microscopy, Giemsa staining, and scanning electron microscopy. Two types of isolated cells were distinguished: large ones which were characterized as superficial cells and smaller ones which seemed to be intermediate basal cells.


Isolation and characterization of a defective measles virus from brain biopsies of three patients in Iran with subacute sclerosing panencephalitis. Three cytopathic strains of subacute sclerosing panencephalitis (SSPE) virus were isolated from brain biopsies of three patients. These strains were isolated and maintained by cocultivation of infected brain cells with fresh Vero cells. The biological characteristics of two strains were studied. It was found that these strains remain cell-associated after repeated cocultivations with Vero cells and produce plaques under fluid medium or tragacanth overlay. The correlation with measles virus was demonstrated by the plaque reduction test as well as by the immunofluorescence test. Large numbers of nucleocapsids were observed in the cytoplasm of infected cells but none in nuclei. Intracerebral inoculation of monkeys, adult guinea pigs, newborn and adult hamsters or mice was followed by acute encephalitis and death.


Simultaneous presence in serum of HBsAg and anti-HBs of different specificities. The simultaneous presence in serum of HBsAg and anti-HBs of different specificities is reported in a patient who had suffered from the third attack of acute viral hepatitis. The antigen was of ayw subtype, while the antibody showed anti-d specificity.


Double contrast myelography.

Double contrast myelography. Myelography was performed with a combination of air and an aqueous myelographic contrast media. Excellent demonstration of the arachnoid sac and cord was possible with double contrast myelography in primates. A technique for double contrast myelography is described. Clinical applications are discussed.


A model for certification of need for long-term-care beds. A model relating bed supply and utilization is presented in the context of the match between need and service, which is controlled by the screening process that allows or denies access to beds. The conventional cost-minimization approach to certification of need, that of seeking to reduce inappropriate use, is contrasted with a service-delivery approach that seeks to promote appropriate use of facilities. The model expresses the quality of the screening process and the sensitivity and specificity of utilization in terms of bed supply, utilization, and need for service, which allows it to be used for needs assessment. The model is applied to data on supply and use of beds in Massachusetts skilled nursing facilities, with screening quality estimated by Monte Carlo methods; the results suggest that need and bed supply are positively associated and that the regional variation in skilled-nursing beds in Massachusetts may reflect real variations in need.


Studies on the ionophorous antibiotics. XI. The artifacts and the degradation products of lysocellin. Lysocellin is a new polyether antibiotic produced by Streptomyces cacaoi var. asoensis and is characterized as a broad spectrum ionophore having a higher complexation affinity for divalent cations than for monovalents and also having an ability to transport biological amines. The structures of two artifacts designated L1 and M1 have been elucidated based on spectral evidence, and the formation mechanism of these compounds was discussed with respect to the reactivity of the antibiotic. In addition, a number of degradation and modified products were prepared in order to examine their biological activity and to utilize as the model compounds for 13C-NMR assignment.


Nocardicin A, a new monocyclic beta-lactam antibiotic III. In vitro evaluation. Nocardicin A, a new monocyclic beta-lactam antibiotic, exerts a comparatively potent antimicrobial activity against gram-negative organisms, especially Pseudomonas aeruginosa, the indole-positive and indole-negative Proteus groups (except Pr. morganii), Serratia marcescens and the Neisseria groups. The in vitro antimicrobial activity of nocardicin A against clinical isolates of Ps. aeruginosa was about twice that of carbenicillin. The mean MICs of nocardicin A for Pr. mirabilis, Pr. rettgeri and Pr. inconstans ranged from 3.13 to 12.5 microgram/ml and were 25 similar to 50 microgram/ml for Pr. vulgaris. Nocardicin A in concentrations of 12.5 similar to 50 microgram/ml inhibited 30 strains (48 percent) of S. marcescens usually resistant to beta-lactam antibiotics. However, nocardicin A had no significant in vitro activity against Staphylococci and Escherichia coli. No cross-resistance was seen between nocardicin A and other beta-lactam antibiotics. This antibiotic was stable to beta-lactamase. The in vitro activity of nocardicin A against Ps. aeruginosa and Pr. mirabilis was greatly influenced by the assay media used. Nocardicin A was bactericidal and appeared to act synergistically with serum bactericidal factors against Ps. aeruginosa and with polymorphonuclear leukocytes against Ps. aeruginosa, E. coli and Pr. vulgaris. The bactericidal activity of nocardicin A against the above 3 organisms, therefore, increased markedly in the presence of fresh serum and polymorphonuclear leukocytes.


Nocardicin A, a new monocyclic beta-lactam antibiotic IV. Factors influencing the in vitro activity of Nocardicin A. Factors influencing the in vitro antimicrobial activity of nocardicin A against Pseudomonas aeruginosa and Proteus mirabilis were investigated. Sodium chloride was identified as a major inhibitor. Some of the amino acids, sugars and divalent cations were found to be minor inhibitors. The presence of potassium phosphates enhanced nocardicin A activity against P. aeruginosa, but antagonized the activity against P. mirabilis.


Environmental synchronizers of squirrel monkey circadian rhythms. Various temporal signals in the environment were tested to determine if they could synchronize the circadian timing system of the squirrel monkey (Saimiri sciureus). The influence of cycles of light and dark, eating and fasting, water availability and deprivation, warm and cool temperature, sound and quiet, and social interaction and isolation was examined on the drinking and activity rhythms of unrestrained monkeys. In the absence of other time cues, 24-h cycles of each of these potential synchronizers were applied for up to 3 wk, and the periods of the monkey's circadian rhythms were examined. Only light-dark cycles and cycles of food availability were shown to be entraining agents, since they were effective in determining the period and phase of rhythmic variables. In the presence of each of the other environmental cycles, the monkey's circadian rhythms exhibited free-running periods which were significantly different from 24 h with all possible phase relationships between the rhythms and the environmental cycles being examined.


Chromosome replication in Myxococcus xanthus. The rates of DNA synthesis during the cell-division cycle were measured in Myxococcus xanthus growing in three different media permitting a twofold variation in doubling time. In all three media, simple DNA cycles were observed. Synthesis of DNA occurred during 85% of the cell-division cycle, independent of generation time, from 5 to 11 h. Cells were observed to contain one bacterial nucleoid at birth that later divided synchronously midway through the cell cycle. Nucleoid segregation appeared to begin before chromosome replication was completed. The DNA content of exponential-phase bacteria was determined to be about 20 +/- 3 X 10(-9) microgram per cell; newborn bacteria contained about 14 +/- 2 X 10(-9) microgram of DNA per cell. Exponential-phase bacteria showed about a 50% increase in DNA in the presence of chloramphenicol (50 microgram/ml). The number of randomly segregating chromosomes present in exponential-phase bacteria was determined by following the fate of prelabeled DNA during outgrowth in nonradioactive media. The results are consistent with a model in which cells are born with exactly one complete unreplicated chromosome. The molecular weight of such a chromosome is about 8.4 +/- 1.2 X 10(9).


Defeat of colicin tolerance in Escherichia coli ompA mutants: evidence for interaction between colicin L-JF246 and the cytoplasmic membrane. Escherichia coli ompA mutants are tolerant to colicin L-JF246. This tolerance can be overcome by a variety of treatments that have as their target the outer membrane or the peptidoglycan layers of the cell envelope. Thus, increasing the concentration of colicin L, releasing lipopolysaccharide from the outer membrane by treatment of intact cells with ethylenediaminetetracetic acid (EDTA), converting cells to spheroplasts by treatment with lysozyme-EDTA or penicillin, or trypsin, treatment of intact cells will result in an increased colicin sensitivity. These treatments alter the outer membrane of ompA mutants and suggest that the altered outer membrane may allow the penetration of at least a portion of the colicin L molecule to a site of action located within this barrier. To substantiate this, we have demonstrated that membrane vesicles prepared from ompA mutants are sensitive to colicin L and that 14C-labeled colicin L binds rapidly to both the outer and inner membrane fractions of the cell.


Peptide utilization in Pseudomonas aeruginosa: evidence for membrane-associated peptidase. A methionine auxotroph of Pseudomonas aeruginosa grew on methionine-containing peptides as a source of the required amino acid. Amino-terminus-blocked peptides would not serve as growth substrates, despite the fact that peptidases active on these blocked peptides were readily detectable in cell extracts. No evidence was found for extracellular enzymes capable of degrading the oligopeptides investigated. The degradative enzymes were not found in the periplasmic space of the cellular envelope. A high proportion of cellular peptidase activity was associated with the particulate (membrane) fraction of the cell lysate.


Effects of detergents on ribosomal precursor subunits of Bacillus megaterium. Cell extracts prepared by osmotic lysis of protoplasts were analyzed by sucrose gradient sedimentation. In the absence of detergents, ribosomal precursor particles were found in a gradient fraction which sedimented faster than mature 50S subunits and in two other fractions coincident with mature 50S and 30S ribosomal subunits. Phospholipid, an indicator of membrane, was shown to be associated with only the fastest-sedimenting ribosomal precursor particle fraction. After the extracts were treated with detergents, all phospholipid was found at the top of the gradients. Brij 58, Triton X-100, and Nonidet P-40 did not cause a change in the sedimentation values of precursors; however, the detergents deoxycholate or LOC (Amway Corp.) disrupted the fastest-sedimenting precursor and converted the ribosomal precursor subunits which sedimented at the 50S and 30S positions to five different classes of more slowly sedimenting particles. Earlier reports on the in vivo assembly of ribosomal subunits have shown that several stages of ribosomal precursor subunits exist, and, in the presence of the detergents deoxycholate and LOC, which had been used to prepare cell extracts, the precursors sedimented more slowly. Our data are consistent with the hypothesis that those detergents selectively modify the structure of ribosomal precursors and lend further support to the hypothesis that the in vivo ribosomal precursor subunits have 50S and 30S sedimentation values. In addition, these data support the idea that the ribosomal precursor particles found in the fast-sedimenting fraction may constitute a unique precursor fraction.


Genetics of the alpha-ketoglutarate dehydrogenase complex of Bacillus subtilis. The enzymatic defects in a number of Bacillus subtilis mutants of the alpha-ketoglutarate dehydrogenase complex lacking activity have been investigated. Mutants in the citK locus, as well as a series of deletions of unknown length covering the citK locus, are deficient in E1 of the complex, alpha-ketoglutarate dehydrogenase, but have normal activities of E2, dehydrolipoyl transsuccinylase, and E3, lipoamide dehydrogenase. The citK mutants and the citL22 mutant show in vitro complementation of alpha-ketoglutarate dehydrogenase complex activity. The citL22 mutant is severely deficient in lipoamide dehydrogenase activity, and, as a result, lacks activity for both the alpha-ketoglutarate and the pyruvate dehydrogenase complexes. Thus, the E3 components of both complexes are identical. The citL22 mutation maps between ura and metC on the chromosome.


Temperature-sensitive nature of the rodB maturation in Bacillus subtilis. The Mg2+ requirement of a morphological mutant of Bacillus subtlis, rodB strain 104 was highly temperature sensitive in the presence of halide or nitrate anions. Likewise the morphological change from rod shapes to spheres was dependent upon temperature, the same anions, and the Mg2+ concentration. The three factors interacted. Other rodB mutants behaved similarly. If the rodB strain 104 in its rod form was treated at high temperatures in the absence of either protein or peptidoglycan synthesis and restored to lower temperatures with the syntheses restarted, a partial temporary change toward cocci occurred. In the absence of halides or in the presence of Cl- but not Br-, the cells increased in volume when they changed from rods to cocci.


Characterization of a Bacillus cereus protease mutant defective in an early stage of spore germination. Temperature-sensitive sporulation mutants of Bacillus cereus were screened for intracellular protease activity that was more heat labile than that of the parental strain. One mutant grew as well as the wild type at 30 and 37 degrees C but sporulated poorly at 37 degrees C in an enriched or minimal medium. These spores germinated very slowly in response to alanine plus adenosine or calcium dipicolinate. During germination, spores produced by the mutant rapidly became heat sensitive, but released dipicolonic acid and mucopeptide fragments more slowly than the wild type and decreased only partially in density while remaining phase white (semirefractile). In freeze-etch electron micrographs, the mature spores were deficient in the outer cross-patched coat layer. During germination, the spore coat changes associated with wild-type germination occurred very slowly in this mutant. Although the original mutant was also a pyrimidine auxotroph, reversion to prototrophy did not alter any of the phenotypic properties discussed. Selection of revertants that germinated rapidly or sporulated well at 37 degrees C, however, resulted in restoratin of all wild-type properties (exclusive of the pyrimidine requirement) including heat-stable protease activity. The reversion frequency was consistent with an initial point mutation, indicating that a protease alteration resulted in production of spores defective in a very early stage of germination.


Structural arrangement of polymers within the wall of Streptococcus faecalis. The structure of the cell wall of Streptococcus faecalis was studied in thin sections and freeze fractures of whole cells and partially purified wall fractions. Also, the structures of wall preparations treated with hot trichloroacetic acid to remove non-peptidoglycan wall polymers were compared with wall preparations that possess a full complement of accessory polymers. The appearance of the wall varied with the degree of hydration of preparations and physical removal of the cell membrane from the wall before study. Seen in freeze fractures of whole cells, the fully hydrated wall seemed to be a thick, largely amorphic layer. Breaking cells with beads caused the cell membrane to separate from the wall and transformed the wall from a predominantly amorphic layer to a structure seemingly made up of two rows of "cobblestones" enclosing a central channel of lower density. Dehydration of walls seemingly caused the cobblestones to be transformed into two bands which continued to be separated by a channel. This channel was also observed in isolated wall preparations treated with hot trichloroacetic acid to remove non-peptidoglycan polymers. These observations are consistent with the interpretation that both peptidogylcan and non-peptidoglycan polymers are concentrated at the outer and inner surfaces of cell walls. These observations are discussed in relation to possible models of wall structure and assembly.


Sequential changes in cell volume distribution during vitamin B12 starvation of Euglena gracilis. During vitamin B12 starvation of Euglena, a new peak appears in the cell volume distribution. Some cells are inhibited at a unique point in the cell cycle between the initiation of DNA synthesis and nuclear division. The mechanism of inhibition of other cells differs.


Enhanced sensitivity to the photodynamic effect of 3,4-benzpyrene in ethylenediaminetetraacetate-treated Escherichia coli. Ethylenediaminetetraacetate treatment of Escherichia coli B enhanced cell sensitivity to the lethal effect of the photodynamic action of 3,4-benzpyrene. The alteration of the shape of the survival curve suggests that such photodynamic action may be modified by membrane barriers to the drug.


Chemotaxis in Pseudomonas aeruginosa.

Chemotaxis in Pseudomonas aeruginosa. A chemotaxis system for Pseudomonas aeruginosa was defined by using the method of Adler. Cells were attracted to compounds in the order ammonium chloride greater than amino acids greater than organic acids. Two sugars were assayed and elicited no response. Comparisons with other model systems are discussed.


Position of the mutation in beta-galactosidase ochre mutant U118. The Escherichia coli lacZ ochre mutant strain U118 was converted to an amber mutant and suppressed with supF, which inserts tyrosine. Enzymatically active beta-galactosidase was isolated. It contained tyrosine at residue number 17 instead of glutamic acid as in wild type.


Monkey glutathione S-aryltransferases. II. Properties of the major enzyme purified from the liver. 1. The molecular and enzymatic properties of the major component (Fraction IV) of glutathione S-aryltransferases [EC 2.5.1.13] purified from the liver of monkey (mainly rhesus monkey) have been investigated. The enzyme had a molecular weight of about 48,000 and was composed of two subunits of apparently identical molecular weight (ca. 24,000) bound to each other non-covalently. Each subunit contained one SH group. The amino acid composition showed characteristic high contents of leucine and glutamic acid residues. No amino-terminal residue was detected by the dansyl method. 2. The enzyme showed a rather broad optimum pH range from 7.5 to 9 with 1,2-dichloro-4-nitrobenzene as a substrate. It was moderately stable below 40 degrees C at pH 7.5. However, it showed an anomalous instability at pH around 4.2. It was reversibly denatured at least partially by urea or guanidine hydrochloride and irreversibly denatured by sodium dodecyl-sulfate. It was significantly inhibited by Zn2+, Cd2+, and Hg2+, and also by benzene hexachloride. It was extensively inactivated by reaction with phenylglyoxal or 2,4,6-trinitrobenzene sulfonate, whereas several SH reagents were without marked effect on the activity under the reaction conditions employed.


Studies on drug-induced lipidosis. VIII. Correlation between drug accumulation and acidic phospholipids. The effect of 4,4'-bis(beta-diethylaminoethoxy)alpha,beta-diethyldiphenylethane (DH) on lipid metabolism in the liver differed considerably in different animal species, humans, monkeys, and rats, because of differences in drug-metabolizing ability. Monkeys retain considerable drug-metabolizing ability as compared with humans, but the DH-hydroxylating activity in monkeys seems to be much lower than in rats. The hydroxyl derivative was the major substance which accumulated in rat liver following the administration of DH, while DH itself and its N-dealkylated substances accumulated in monkey liver. N-Dealkylated substances were also observed in human liver, but the amount was much smaller than in monkeys. Bis(monoacylglyceryl)phosphate (BMGP), which is characteristic of this kind of drug-induced lipidosis, did not increase as much in monkey liver as in human liver, but a marked increase in phosphatidyl inositol (PI) was observed in monkey liver during administration of DH. The concentration of acidic phospholipids (BMGP + PI) in liver showed a close correlation with the accumulation of the drug (DH + its metabolites), irrespective of species differences. Among subcellular particles isolated from a monkey liver following administration of DH, the crude mitochondrial fraction, including lysosomes, was richest in BMGP.


Biosynthesis of a prostaglandin by a plant enzyme. Lipoxygenase-2 from soybeans catalyzes the oxygenation of arachidonic acid to form significant amounts of a prostaglandin product. Results obtained with the dithionite-reduced derivatized product upon combined gas chromatography-mass spectroscopy are consistent with structures of the stereoisomers of 9,11,15-trihydroxyprosta-5,13-dienoic acid. A portion of the material formed by reduction of the enzyme product with dithionite reacts positively in a radioimmunoassay against rabbit anti-prostaglandin F2alpha antibody. This appears to be the first report of the synthesis of a prostaglandin by a non-animal enzyme.


Purification and characterization of human erythrocyte purine nucleoside phosphorylase and its subunits. Purine nucleoside phosphorylase (EC 2.4.2.1; purine nucleoside:orthophosphate ribosyltransferase) from fresh human erythrocytes has been purified to homogeneity in two steps with an overall yield of 56%. The purification involves DEAE-Sephadex chromatography followed by affinity chromatography on a column of Sepharose/formycin B. This scheme is suitable for purification of the phosphorylase from as little as 0.1 ml of packed erythrocytes. The native enzyme appears to be a trimer with native molecular weight of 93,800 and the subunit molecular weight of 29,700 +/- 1,100. Two-dimensional gel electrophoresis of the purified enzyme under denaturing conditions revealed four major separable subunits (numbered 1 to 4) with the same molecular weight. The apparent isoelectric points of subunits 1 to 4 in 9.5 M urea are 6.63, 6.41, 6.29, and 6.20, respectively. The different subunits are likely the result of post-translational modification of the enzyme and provide an explanation of the complex native isoelectric focusing pattern of purine nucleoside phosphorylase from erythrocytes. Three of the four subunits are detectable in two-dimensional electrophoretic gels of crude hemolysates. Knowing the location of the subunits of purine nucleoside phosphorylase in a two-dimensional electropherogram allows one to characterize the purine nucleoside phosphorylase in crude cell extracts from individuals with variant or mutant purine nucleoside phosphorylase as demonstrated in a subsequent communication. Partial purification of the phosphorylase from 1 ml of erythrocytes on DEAE-Sephadex increases the sensitivity of detection of the subunits to the 0.3% level.


Development of action potential in larval muscle fibers in Drosophila melanogaster. In the presence of tetraethylammonium or barium ions, the larval muscle fibers of Drosophila melanogaster were found to produce an all-or-none action potential operated by the calcium channels. The development of this distinctive membrane property during the maturation of muscle cells was studied by measuring the maximum rate of rise of the action potential in the larval muscle fibers at different stages of development from the sixteenth to ninety-sixth hours after hatching. The value increased significantly with age until a peak was reached at the sixty-fourth hour, although it became lower again as puparium formation neared at about the ninety-sixth hour. This suggests that during larval development the muscle fibers develop the ability to generate an action potential due to an inward current through the calcium channels, although the ability became lower at the later stage of larval development.


Fine structure and RNA synthesis of Tetrahymena during cytochalasin B inhibition of phagocytosis. Cytochalasin B inhibits the formation of normal-sized food vacuoles in Tetrahymena but the cells do not starve. Treated cells differ from starved cells in that they retain a high rate of incorporation of tritiated uridine. Large numbers of smaller vacuoles, about 1 micrometer in diameter, are formed, presumably by pinocytic activity of the cytopharyngeal membrane. This effect may perhaps be due to interference with the mechanism by which food vacuoles are sealed off at the cytostome, in which microfilaments may participate. Inhibited organisms may form tubes continuous with the cytopharynx instead of separate food vacuoles. It is not clear, however, why the formation of the small vacuoles is resistant to the drug.


The fixation of nuclei in glutaraldehyde. The production of an artifactual network in the nuclear sap of the salivary glands of Drosophila has been investigated. Mechanical stress to the cells, 3% glutaraldehyde containing more than 4 mM calcium, tannins, or cacodylate buffer or whose temperature is above 10 degrees C all enhance this artifactual effect. Over the range pH 6.65-7.4 there is no significant effect of pH. The inclusion of sucrose, and adenine nucleotides or other 'chelating' agents in the fixative reduces the effect, especially at low temperature (6-10 degrees C). These influences are additive and can abolish the artifactual effect. So too can 8% glutaraldehyde, but this is irrespective of temperature.


Thermal transitions in the adhesiveness of HeLa cells: effects of cell growth, trypsin treatment and calcium. The intercellular adhesiveness of density-inhibited (D.I.) and fast-growing (F.G.) HeLa cells and of trypsin-treated preparations of these, has been measured at temperatures between 37 and 6 degrees C. In EDTA-containing buffer medium, F.G. cells differ from D.I. cells in that only the former display an increase in adhesiveness below 30 degrees C. This increase is prevented by previous treatment with trypsin. The presence of Ca2+ in the buffer medium causes a narrowing of the thermal transition region of intact F.G. cells. On intact D.I. cells Ca2+ causes an increase in adhesiveness at temperatures below 20 degrees C. Previous trypsinization of F.G. cells diminishes the effect of subsequent Ca2+ addition. The adhesiveness of trypsinized D.I. cells is indifferent to changes in temperature in Ca2+-containing buffer medium. The results are considered evidence for the occurrence of a phase transition in the glycoprotein domains of the plasma membrane of fast-growing cells. The transition is influenced by growth rate, trypsinization and Ca2+. The fluidity of the membrane glycoproteins is considered to be higher on density-inhibited cells than on fast-growing cells. No phase transition could be detected after incorporation of the fluorescent compound 1,6-diphenyl 1,3,5-hexatriene into the lipid domain of the plasma membrane. The fluidity of the membrane lipids is lower on density-inhibited cells than on fast-growing cells.


Synthesis of proteoglycans by suspension and monolayer cultures of adult chondrocytes and de novo cartilage nodules-the effect of hyaluronic acid. Chondrocytes were isolated from adult laryngeal cartilage by an enzymic procedure that included 6 h digestion with collagenase. The level of 35SO4(2-) incorporation into cetylpyridinium chloride-precipitable material by these cells depended upon the subsequent culturing conditions. Suspension cultures incorporated more 35SO4(2-)/cell than monolayer cultures. Hyaluronic acid in the medium inhibited 35SO4(2-) incorporation only when the cells were in primary suspension cultures. It had no effect on monolayer cultures, or monolayers organized into nodules, or suspension cultures derived from monolayers. Mild pretreatment with EDTA, however, rendered these susceptible to hyaluronic acid inhibition. In contrast EDTA abolished the inhibitory effect of hyaluronic acid on primary suspension cultures. Oligosaccharides, derived from hyaluronidase digestion of hyaluronic acid that were larger than decassaccharide, had some inhibitor effect on 35SO4(2-) incorporation by monolayer cultures. The total 35SO4(2-) incorporation was less in primary suspension cultures of chondrocytes isolated after 12 h than after 6 h digestion of cartilage and the inhibition by hyaluronic acid was also less. These differences persisted during 12 days of culture. It is suggested that the method of isolating chondrocytes and subsequent culture conditions may modify the cell surface and mask or abolish specific binding sites for hyaluronic acid.


Dissociation of prolactin responsiveness to TRH and chlorpromazine in women with isolated gonadotropin deficiency. The hormonal response to luteinizing hormone releasing hormone (LHRH) thyrotropin releasing hormone (TRH) and chlorpromazine has been evaluated in eleven female subjects with the syndrome of isolated bihormonal gonadotropin deficiency (IGD). Following LHRH, all subjects had elevations of both LH and FSH, but the gonadotropin responses were attenuated relative to those observed in normal female subjects studied in the early proliferative phase of the cycle. Similarly, peak TSH levels after TRH were significantly less in subjects with IGD relative to normal controls. Basal prolactin levels were low in the patient group. Prolactin levels following TRH increased at least two-fold in control subjects and in the group with IGD. Conversely, chlorpromazine failed to induce elevations of prolactin in eight of nine females with IGD.


The clinical and biochemical implications of pyruvate carboxylase deficiency. A 10 month old female infant was evaluated for severe lactic acidosis. Clinically she was well nourished and had a substantial amount of adipose tissue despite recurrent episodes of acidosis. Her psychomotor development was retarded, her movements were dystonic and generalized seizures punctuated her course. Metabolic abnormalities included elevated blood concentrations of lactate, pyruvate, beta-hydroxybutyrate, acetoacetate, alanine, proline and glycine, decreased blood concentrations of glutamine, aspartate, valine and citrate, and intermittent elevations of serum cholesterol. A trial on a high-fat diet worsened the clinical condition and intensified the ketoacidosis and hyperalaninemia. Analysis of hepatic tissue obtained by open biopsy revealed increased concentrations of lactate, alanine, acetyl-CoA and other short-chain acyl-CoA esters, and decreased concentrations of oxaloacetate, citrate, alpha-ketoglutarate, malate and aspartate. The blood and tissue metabolic perturbations reflected a deficiency of hepatic pyruvate carboxylase. The apparent Km of hepatic citrate synthase for oxaloacetate was 4.6 micrometer. Calculated tissue oxaloacetate concentrations were 0.50--0.84 micrometer suggesting that tricarboxylic acid cycle activity was severely limited by the decreased availability of this substrate. An iv glucose tolerance test resulted in the paradoxical synthesis of ketone bodies. This observation, coupled with the intermittent hypercholesterolemia and the increased tissue acetyl-CoA concentrations, suggests that pyruvate carboxylase is important in modulating the fractional distribution of intracellular acetyl-CoA between the tricarboxylic acid cycle, the beta-hydroxy-beta-methyl-glutaryl-CoA cycle (and the synthesis of cholesterol and ketone bodies), and fatty acid synthesis. Treatment in future cases might be directed toward increasing tissue concentrations of oxaloacetate.


Solid-phase competitive-binding radioimmunoassay for detecting antibody to the M antigen of histoplasmin. A radioimmunoassay (RIA) was designed and compared with complement fixation and immunodiffusion tests for their relative ability to detect antibodies in sera of histoplasmosis patients. M antigen, purified from histoplasmin, was fixed to microtiter wells as the solid phase, and specific rabbit 125I-labeled anti-M globulin was the source of indicator antibodies. The optimal concentrations for the competitive-binding assay were 1.6 ng per well for M antigen and 650 ng per well for the 125I-labeled anti-M globulin. A panel of sera from 29 histoplasmosis patients and from patients with other mycoses was screened for RIA activity and in complement fixation and immunodiffusion tests that used histoplasmin and Histoplasma capsulatum yeast-form antigens. The sera of 22 histoplasmosis patients reacted in the RIA, 21 in the complement fixation, and 16 in the immunodiffusion tests. Sera of patients with other mycotic infections did not react in the RIA, with the exception of those of one blastomycosis patient and one candidiasis patient. The RIA could be modified to quantitate M antigen; as little as 125 pg could be detected. The evaluation of this panel of histoplasmosis patients' sera showed that the RIA was about equivalent in sensitivity to the complement fixation test. Some advantages of the RIA over the complement fixation test were that RIA was less prone to cross-reactions and gave better quantitation of low-titered sera. The RIA was a 1-day test, was not hindered by the anti-complementary activity of some sera, and could be modified to quantitate minute amounts of M antigen.


La Crosse virus soluble cell culture antigen. A virus-free soluble antigen, obtained by ammonium sulfate precipitation of the supernatant fluids of La Crosse virus-infected BHK-21 cell cultures, was more reactive and more specific than infected suckling mouse brain antigen when compared by immunodiffusion and counterelectrophoresis tests. By complement fixation tests, the antigen was cross-reactive with heterologous California group arbovirus hyperimmune mouse ascitic fluids, but to a lesser degree than was the standard sucrose-acetone-extracted infected suckling mouse brain antigen. The major virion nucleocapsid protein of La Crosse virus was found by polyacrylamide gel electrophoresis to be the soluble antigen protein responsible for precipitation in immunodiffusion and counterelectrophoresis tests.


Improved chicken embryo cell culture plaque assay for scrub typhus rickettsiae. The plaque technique for three strains of Rickettsia tsutsugamushi in chicken embryo cell cultures was greatly improved by modifying the trypsinizing procedure and employing homologous chicken serum in the overlay medium.


The ultrasonic characteristics of the renal pelvicalyceal echo complex. The pelvicalyceal echo complex has a characteristic appearance as an echo-filled cylinder in which minimal sonolucency may normally be seen. Alteration in the pelvicalyceal echo complex is a helpful guide in the diagnosis of a variety of pathological processes involving the kidneys. Reliable diagnosis of moderate and marked hydronephrosis may be made by the appearance of uniformly sized, sonolucent collections in the pelvicalyceal echo complex which show communication with the renal pelvis. An adjacent simple renal cyst produces a sharply defined crescent shaped deformity of the pelvicalyceal echo complex, and scalloping is seen with multiple impinging renal cysts. Gross deformity of the central echo complex is seen with polycystic kidney disease, as well as in neoplasia. Edema and infiltrating hypernephroma may result in diminution of the pelvicalyceal echo complex. Amputation or a "V"-shaped splitting of the pelvicalyceal echo complex may be seen with neoplasia.


Sonographic demonstration of intrauterine contraceptive devices. Ultrasonic examination of the uterus is the investigation of choice in the initial localization of an IUCD within the uterine cavity. The use of compound B scanning in conjunction with A scans has been completely reliable to date. Radiography should be reserved for those patients in whom an expected device is not shown within the uterine cavity, and for those in whom the determination of the type of device is important and the scan is equivocal.


The ultrasonic demonstration of gastric lesions. Fifteen cases of circumferential gastric antral disease identified by upper gastrointestinal (GI) series over a 12-month period were studied by a gray-scale ultrasound. In 93 per cent of the patients studied, a characteristic ovoid anechoic area containing a dense central collection of echoes was identified. The abnormality was located in the midline or slightly to either side, just dorsal to the inferior margin of the liver on longitudinal sections. The lesions studied included primary and secondary malignancy, lymphoma, corrosive gastritis, and granulomatous disease of childhood. In each instance a barium study of the stomach confirmed the location of the lesion. A prospective study of 50 normal individuals revealed one patient in whom the characteristic appearance was found at ultrasound and who subsequently had a normal upper GI series. Thus, although the ultrasound appearance of antral thickening may rarely be found in normal individuals, this characteristic appearance should strongly suggest gastric pathology, necessitating an upper GI series.


Staging of bladder tumors by means of transrectal ultrasonography. Transrectal ultrasonography is a specially devised technique to obtain clear tomograms of the intrapelvic organs, such as the prostate, urinary bladder and seminal vesicles. We used this technique to evaluate 64 patients with bladder tumors. The results were compared with histological findings after exploratory surgery. Ultrasonograms of these tumors were estimated on the basis of bladder wall irregularity, bladder deformity, bladder capacity and extravesical spread of tumor. Each tumor stage showed characteristic ultrasonic features. Extravesical spread of tumor, which was observed in 67 percent of stage D tumors, proved to be an important finding to distinguish stage D tumours from other tumor stages. We suggest these features as diagnostic criteria for ultrasonic staging of bladder tumors.


Gray scale echography with simultaneous A mode presentation. A new apparatus for high resolution ultrasonograms. A diagnostic ultrasonic system has been developed for simultaneous display of quantitative A mode and gray scale B mode presentations. An independent calibrator and preset amplification levels allow the apparatus to be calibrated for precise, repeatable examinations. A wide range of echo amplitude information is available to the clinician by the use of selective logarithmic signal compression. Distance measurements in the eye and the orbit are made to 0.1 mm precision and are displayed numerically on the control panel. The newer signal processing techniques provide high-resolution imaging of ophthalmic structures over a wide system bandwidth. Sensitivity has been improved to image structures heretofore not observable with ultrasonic instruments. This technique is the basis for recently initiated clinical studies.


Reliability of echocardiography in quantitating pericardial effusion: a prospective study. A method of estimating the volume of pericardial effusion by echocardiography has used the difference between the cubed diameters at end-diastole of the pericardium and epicardium. To evaluate the reliability of this technique in quantitating the volume of pericardial effusion in a prospective study, 22 echocardiograms were obtained in six patients before and after 11 separate pericardiocenteses. The correlation coefficient between the actual volume of aspirated pericardial effusion and the echocardiographically estimated volume of aspirated pericardial effusion was r = 0.27 (P not significant). The volume of pericardial effusion aspirated was overestimated or underestimated by echocardiography by more than 100 ml in seven of 11 estimations (64 percent) and by more than 150 ml in five of 11 estimations (45 percent). Therefore, although echocardiography is the procedure of choice in diagnosing the presence of pericardial effusion, it is not an entirely accurate method of quantitating the volume of pericardial effusion. However, echocardiography can differentiate a large effusion from a moderate or small effusion.


In search of the elusive benign cystic ovarian teratoma: application of the ultrasound "tip of the iceberg" sign. The benign cystic ovarian teratoma (dermoid) has distinctive ultrasonic features that differentiate it from other adnexal masses. The most frequent sign is the presence of highly reflective irregular solid components within a fluid-containing adnexal mass. In vitro scans of surgical specimens have shown that the strongly reflective echo pattern is caused by the hair and sebum within the dermoid. Calcium is often sparse or absent. Acoustic shadowing from the hair may totally obscure the back wall of a large, clinically evident mass; hence the term "tip of the iceberg" sign. Recognition of this sign reduces the possibility of a false negative interpretation and confusion with a "bowel gas pattern."


Omental band: new sign of metastasis.

Omental band: new sign of metastasis. Lymphomatouscellular infiltration of the greater omentum has been visualized by ultrasound as a uniformly thick, hyposonic, band-shaped structure adjacent to the anterior and lateral walls of the abdomen, following the contour of the abdominal convexity, and containing low-level, nonstructured internal echoes. From this finding, diffuse metastatic disease to the omentum can probably be inferred with a high degree of reliability.


Echocardiographic manifestations of flail aortic valve leaflets. Six patients with aortic regurgitation secondary to flail aortic valve leaflet syndrome were studied echocardiographically, angiographically, surgically, and pathologically. The etiology of flail aortic leaflet was myxomatous degeneration in four patients and bacterial endocarditis in two. The echocardiographic manifestations of flail aortic valve leaflet are diastolic fluttering of the aortic cusp echoes, abnormal systolic aortic leaflet movement, and abnormal diastolic fluttering echoes in the left ventricular outflow tract. Diastolic fluttering of the aortic valve leaflets is specific echocardiographic evidence of aortic regurgitation due to flail aortic leaflets, and demonstration of diastolic fluttering of the aortic leaflets and diastolic fluttering echoes in the left ventricular outflow tracts is direct evidence of flail and prolapsed aortic leaflets.


Ultrasound diagnosis of intramural intestinal hematoma. Intramural intestinal hemorrhage in hemophilia has been diagnosed by ultrasound. The sonographic pattern consists of a tubular anechoic mass containing a core of strong echoes. Recognition of intestinal hematoma is important because of its bearing on treatment and prognosis. To our knowledge, this is the first reported case.


Interrelationship of Chlamydia trachomatis and other pathogens in the female genital tract. The isolation of Neisseria gonorrhoeae, Chlamydia trachomatis, Trichomonas vaginalis, and Candida albicans in the female genital tract was studied in 1323 patients attending a venereal disease clinic. Disruption of the cell monolayers use for the isolation of C. trachomatis was significantly associated with the presence of T. vaginalis; this effect was markedly reduced by the addition of vancomycin to gentamicin and amphotericin B in the transport and growth media. The only significant positive association was the more frequent isolation of C. trachomatis in the presence of N. gonorrhoeae. There was a negative association between N. gonorrhoeae and C. albicans and between T. vaginalis and C. albicans, the fungus being isolated significantly less frequently when these microorganisms were present.


Parathyroid carcinoma in familial hyperparathyroidism. Two families with hereditary hyperparathyroidism are described. One member of each family developed a parathyroid carcinoma. In one case this recurred locally and metastasised. This patient showed hyperplasia of one of the three other parathyroid glands. It is possible that the different parathyroid lesions found in familial hyperparathyroidism may be the result of a progression from hyperplasia to formation of benign or malignant tumours. The remainiing hyperplastic glands may be suppressed by hypercalcaemia. There was no evidence of multiple endocrine neoplasia in either family. Three members of a first family had ichthyosis and both affected members of the second had tumours of the jaw, one of which was an ossifying fibroma, suggesting a possible association of these conditions with familial hyperparathyroidism.


A double-blind comparison of meptazinol with pentazocine and placebo in cancer pain. Two consecutive double-blind, crossover, single-administration trials have shown that intramuscular meptazinol, a new opioid-antagonist drug, is superior to placebo and equivalent ot pentazocine in relieving chronic pain due to cancer.


The locus and cytoarchitecture of the projection areas of the olfactory bulb in Macaca mulatta. A study was made of the normal and experimental anatomy of the olfactory system of the young adult male rhesus monkey. The cytoarchitecture of the central olfactory areas was studied with cell and fiber stains, while the extent and pattern of the projections of the olfactory bulb were determined by the Fink-Heimer and autoradiographic methods. The brain of one animal that had sustained damage to the olfactory bulb two days prior to sacrifice, and of one that had a transection of the olfactory tract ten days prior to sacrifice, were processed with the Fink-Heimer technique. The first of these and four others received injections of 3H-proline or 3H-leucine into the olfactory bulb, and following a survival period of 18 hours, or 2, 4, 12, or 20 days, their brains were processed with the autoradiographic technique. The results were the same for both experimental methods and for all survival periods. The projections of the olfactory bulb in this microsmatic animal are entirely ipsilateral. All of the structures that receive direct olfactory afferents have a laminar organization except for the anterior olfactory nucleus, which is laminated only in its anterior, peduncular, portion. While the olfactory bulb projects to the entire extent and depth of the anterior olfactory nucleus, the olfactory afferents of all other structures are confined to layer IA of the plexiform layer. These structures are: all divisions of the olfactory tubercle; the frontal and temporal prepiriform cortices; the oral, medial, and dorsal divisions of the superficial amygdaloid nucleus; and polar and anterior entorhinal cortex. The rhesus monkey does not have a recognizable accessory olfactory bulb, and no projections were seen to one of its targets, the nucleus of the stria terminalis. Also, no projections were seen to the taenia tecta or the ventral division of the superficial amygdaloid nucleus. With these exceptions, the projections of the olfactory bulb in the rhesus monkey are similar to those in macrosmatic species.


A brainstem atlas of catecholaminergic neurons and serotonergic perikarya in a pygmy primate (Cebuella pygmaea). The present paper provides a brainstem atlas showing the distribution of catecholaminergic cells and processes, as well as serotonergic perikarya, in the pygmy marmoset. The findings revealed by the Falck and Hillarp histofluorescence method conform in essential details to what has been described in rodents. These and other comparative data indicate that a prototypical pattern of organization of aminergic systems has been retained in the evolution of primates.


Excitation of marginal and substantia gelatinosa neurons in the primate spinal cord: indications of their place in dorsal horn functional organization. Electrophysiological recordings were made from superficial parts of the spinal dorsal horn in monkeys, using dye-filled micropipette electrodes to permit iontophoretic marking of the recording sites for subsequent histological recovery. Focal field potentials and unitary activity evoked by dorsal root volleys including slowly-conducting components (both myelinated and unmyelinated) were found in the posteromarginal zone and the substantia gelatinosa (SG). Unitary potentials identified as being of the type recorded from cellular regions were separated into categories according to which group of slowly-conducting fibers and which kinds of cutaneous stimulation evoked the discharge. Recording locations for units excited by volleys in myelinated fibers conducting under 35 m/sec, by the types of skin stimulation activating either high-threshold mechanoreceptors (nociceptors) or cooling thermoreceptors, and giving no evidence of suprathreshold C-fiber excitation were centered on the posteromarginal zone. In contrast, recording loci for units exhibiting a strong C-fiber excitation and responses to cutaneous stimulation known to effectively excite C-fiber polymodal nociceptors or C-mechanoreceptors were centered in the SG. There appeared varying degrees of convergence of primary afferent input to the neuronal units, although most showed substantial specificity in their afferent excitation. On the bases of these results and consideration of existing morphological data, it is proposed that the marginal zone is a major synaptic termination region for the afferent fibers from high-threshold mechanoreceptors, cooling thermoreceptors, and perhaps other receptors with fine myelinated peripheral fibers. The SG, on the other hand, is suggested to be the terminal region for all types of unmyelinated primary afferent sensory neurons, and to have the complex role of integrating and distributing this input.


Some claustro-cortical connections in the cat and baboon as studied by retrograde horseradish peroxidase transport. 1. The mammalian Claustrum (Cl) is a convergent multisensory structure of unknown function, and disputed ontogenetic origin. Its cortical projections, hitherto unknown, have been studied in cat and baboon by means of the horseradish peroxidase (HRP) technique. HRP was injected into the gyrus proreus (frontal eye field) of cats, and separately into the frontal eye fields, visual areas, and motor-premotor areas of the baboon cortex. 2. Differential retrograde transport to the Cl was demonstrated, such that in the cat the ipsilateral dorsal Cl was shown to be the principal origin of claustroproreate projections. In the baboon, the whole Cl projects onto area 8, while only the posteroventral part of the nucleus sends efferents to the visual cortex. The projection to the motor and premotor areas is present, but does not seem to be "essential." 3. Discussion of the physiological literature, together with anatomical evidence of reciprocal cortico-claustral projections to closely similar regions of the Cl lead to the suggestion that the Cl is concerned with the integration of messages subserving visually-directed movements. Some other functional implications are also discussed.


Immunologic studies on drug addiction: III. Antibodies reactive with cocaine metabolites and their use for drug detection. Antisera which react with the major metabolites of cocaine have been prepared in rabbits and a hemagglutination-inhibition (HI) test which detects these metabolites in urine or serum in concentrations of 1 ng/ml is described. A comparison of this test with alternate detection procedures shows excellent agreement at comparable sensitivity levels. HI tests on simian and human biological specimens suggest that the use of cocaine remains detectable for at least 3 days after administration of a minimal pharmacologically active dose. Combination of physical separation of drug metabolites with immunoassay procedures indicates that benzoylecgonine and ecgonine are the immunoreactive cocaine metabolites found in human urine. While it was possible to apply the HI test at maximal sensitivity to human sera and to murine or simian urine specimens, interference was encountered with some human urine specimens. Preliminary data suggest that by reducing the sensitivity of the test to a threshold of 200 ng/ml this interference can be overcome.


Radioimmunoassay for antigliadin-antibodies using 14C-labelled gliadin. A sensitive radioimmunoassay for antibodies to gliadin has been developed. Gliadin from wheat gluten was labelled with [1-14C]acetic anhydride to a specific activity of 2.6 X 10(6) dpm/mg. Immunological evidence is presented that the antigen was not essentially altered by the labelling procedure. Experimentally-induced antigliadin antibodies or sera of patients with coeliac disease (CD) were reacted with labelled gliadin and the immune complexes formed precipitated by antiglobulin. Precipitating antibodies were determined by incubating CD sera with labelled gliadin and measuring the radioactivity in precipitates formed without the addition of second antibody. Comparison with other methods for the detection of antigliadin antibodies, including immunoelectrophoresis, immunodiffusion and passive hemagglutination indicated that total and precipitating antibodies were determined only by RIA. The assay also provides information on the immunoglobulin class of antigliadin-antibodies present in sera of patients with coeliac disease.


The use of denaturing conditions for gel diffusion of insoluble epidermal proteins. Modifications of the classical technique for double diffusion in agar plates are presented. These procedures allow insoluble proteins to be dissolved in a variety of denaturing solvents and antibody production specificity to be monitored by the appearance of precipitin lines in agarose gels as in the conventional Ouchterlony method. Results obtained using the insoluble proteins of bovine epidermis are summarized.


Decreased virulence of gentamicin-resistant strains of Pseudomonas aeruginosa in a rat model. Previous studies have reported that GR strains of Pseudomonas aeruginosa were less likely to cause serious infections. In the present study, this clinical observation of decreased virulence of GR strains of P. aeruginosa was documented in Andriole's rat model. A significantly higher mortality in rats was noted with GS strains of P. aeruginosa (MIC less than 10 microgram/ml) compared to moderately resistant strains (MIC 12.5 to 312 microgram/ml) and highly resistant strains (MIC greater than 312 microgram/ml). Slower growth rates of GR strains of P. aeruginosa were observed in both the lag and the log phases as compared to GS strains. Average counts of in vivo quantitative blood cultures with the use of GR strains were lower at 30 min, 8 hr, and 24 hr compared to those with GS strains. The mortality in mice injected intraperitoneally with CFS's from GS strains (MIC 0.78 to 3.125 microgram/ml) was significantly higher at 8 and 24 hr than those injected with supernatants from GR strains (MIC 25 to 400 microgram/ml). Human neutrophils killed GR and GS strains of P. aeruginosa equally. Decreased virulence of GR strains of P. aeruginosa may be due in part to slower growth rates and to a decreased ability to produce heat-labile toxic components.


Study of platelet aggregation in vivo: IV Effect of pyrimido-pyrimidine derivatives. Two pyrimido-pyrimidine derivatives (RA 233  and VK 744) were found effective aggregation inhibitors in vitro of human and monkey platelets and in vivo in stumptailed monkeys and chimpanzees. They also inhibited thrombotic occlusion of metal screens inserted into arteriovenous shunts.


Study of platelet aggregation in vivo: V effect of the antiserotonin agent cyproheptadine. The antiserotonin agent cyproheptadine (Periactin) was shown to be an effective antagonist of in vivo platelet aggregation induced in monkeys not only with serotonin but also with ADP.


[Ultrastructural features of ureter-mesenchyme interaction, during nephrogenic induction in Amphibia]. Transmission electron microscopy was used to study the interface between the ureter and the mesonephric blastema. When the blastema is induced in a functional organ, numerous pseudopodial protrusions of the ureter and of the mesonephric cells lead to a close appostion between the two structures. The presence of squamous material, perhaps of a mucopolysaccharidic nature, with a network of collagen fibres is always found on this interface. These particular processes seem to play a role as a morphological basis for the phenomenon of induction. The importance of the extracellular matrix is discussed. Neither the occurrence of zones of close contact, reported by several authors, in other species, nor the fusion of membranes has ever been observed in Amphibia.


Total parenteral nutrition: a guide to therapy in the adult. Total parenteral nutrition (TPN), often referred to as intravenous hyperalimentation, is a complex technique for parenteral feeding that can be lifesaving. A basic knowledge of the theory behind, indications for, and hazards of TPN can help the practitioner determine which of his patients will benefit from this procedure. In the community hospital, where a skilled hyperalimentation team is not available, TPN can be managed safely by a physician, pharmacist and floor nurse if there is rigid adherence to a strict protocol. This paper presents the basic theory, indications, and contraindications associated with TPN, and details a protocol for administering total parenteral nutrition to the adult, hospitalized patient (Appendices 1, 2).


Endocrine control of vitellogenesis in Drosophila melanogaster: effects of the brain and corpus allatum. The endocrine control of yolk deposition in Drosophila melanogaster was studied by ligation and transplantation techniques. Endocrine events associated with the initiation of vitellogenesis were found to be synchronized with eclosion rather than the completion fo adult development. Decapitation experiments showed that a cephalic event occurring at about the time of eclosion is necessary for each animal to initiate vitellogenesis. The morphogenetic effect of the head could be replaced by a juvenile hormone analog (JHA). In addition to the cephalic event, a thoracic factor is required for each follicle to initiate vitellogenesis, since preparation of isolated abdomens before 16 hours after eclosion prevented vitellogenesis. In abdomens isolated after this time, no early vitellogenic stages were formed. The suppression of vitellogenesis in isolated abdomens was reversed by implanting corpora allata or by treating these preparations with JHA, but not by implanting corpora cardiaca. Ovaries that were artificially induced to mature by treating isolated abdomens with JHA still displayed the normal complement of ovarian proteins after electrophoresis in polyacrylamide gels. These results show that a circadian clock triggers vitellogenesis via a cephalic signal at eclosion, which in turn triggers events in the thorax or abdomen. The cephalic signal can be superseded by juvenile hormone, whose presence is necessary for each follicle to become vitellogenic.


Electrical and adaptive properties of rod photoreceptors in Bufo marinus. I. Effects of altered extracellular Ca2+ levels. The effects of altering extracellular Ca(2+) levels on the electrical and adaptive properties of toad rods have been examined. The retina was continually superfused in control (1.6 mM Ca(2+)) or test ringer's solutions, and rod electrical activity was recorded intracellularly. Low-calcium ringer's (10(-9)M Ca(2+)) superfused for up to 6 min caused a substantial depolarization of the resting membrane potential, an increase in light-evoked response amplitudes, and a change in the waveform of the light-evoked responses. High Ca(2+) ringer's (3.2 mM) hyperpolarized the cell membrane and decreased response amplitudes. However, under conditions of either low or high Ca(2+) superfusion for up to 6 min, in both dark-adapted and partially light-adapted states, receptor sensitivity was virtually unaffected; i.e., the V-log I curve for the receptor potential was always located on the intensity scale at a position predicted by the prevailing light level, not by Ca(2+) concentration. Thus, we speculate that cytosol Ca(2+) concentration is capable of regulating membrane potential levels and light-evoked response amplitudes, but not the major component of rod sensitivity. Low Ca(2+) ringer's also shortened the period of receptor response saturation after a bright but nonbleaching light flash, hence accelerating the onset of both membrane potential and sensitivity recovery during dark adaptation. Exposure of the retina to low Ca(2+) (10(-9)M) ringer's for long periods (7-15 min) caused dark-adapted rods to lose responsiveness. Response amplitudes gradually decreased, and the rods became desensitized. These severe conditions of low Ca(2+) caused changes in the dark-adapted rod that mimic those observed in rods during light adaptation. We suggest that loss of receptor sensitivity during prolonged exposure to low Ca(2+) ringer's results from a decrease of intracellular (intradisk) stores of Ca(2+); i.e., less Ca(2+) is thereby released per quantum catch.


The regulation of stable RNA synthesis in the blue-green alga Anacystis nidulans: effect of leucine deprivation and 5-methyltryptophan inhibition. The expression of phenomena associated with the bacterial function controlling RNA synthesis was studied in leucine-deprived or 5-methyltryptophan-treated cultures of Anacystis nidulans. Both procedures retarded cell growth, RNA and protein accumulation, elicited the accumulation of high intracellular concentrations of guanosine 5'-diphosphate-3'-diphosphate(ppGpp)and guanosine5'-triphosphate-3'-diphosphate(pppGpp),and promoted a regime of non-coordinate synthesis of stable and messenger RNA. The rate of polymerization of nascent RNA chains did not appear to be retarded in the growth-limited cultures.


Phenotypic changes in the resistance of Neisseria gonorrhoeae to killing by normal human serum. Gonococci adapted to growth in chambers implanted subcutaneously into guinea pigs are resistant to killing by human serum. This resistance is lost after a few generations in vitro both in culture medium and in fluid taken from guinea-pig chambers. The rate of loss is too rapid to occur by mutation and selection. Furthermore, the resistance is regained after a few generations when bacteria from the first in vitro culture are inoculated back into guinea-pig chambers in vivo. Hence the loss of serum resistance in vitro and the gain in vivo are probably due to phenotypically controlled events. Such events could be important in the pathogenicity of Neisseria gonorrhoeae.


Gold encephalopathy.

Gold encephalopathy. Encephalopathy is described developing in a 42 year old man who was receiving gold treatment for rheumatoid arthritis.


The electroencephalogram in neglect.

The electroencephalogram in neglect. The extent of electroencephalographic (EEG) abnormalities in 23 consecutive patients with unilateral neglect was compared with the extent of EEG abnormalities in 20 matched aphasic controls. In the neglect group, 22 patients showed EEG slowing that extended over the entire hemisphere. In the control group only 8 patients showed similar hemispheric slowing. This study demonstrates that patients with unilateral neglect from discrete lesions may have EEG slowing in regions of the hemisphere that are remote from the lesion. This study also provides support for the hypothesis that neglect is a unilateral arousal defect.


Asphyxia-induced disaggregation of cerebral polyribosomes in rhesus monkey fetuses. The effects of 3 hours of controlled intrauterine asphyxia (acidotic hypoxia) on the sedimentation patterns of cerebral polyribosomes and on polyribosome supported in vitro protein synthesis were examined in 16 term monkey fetuses. Three outcomes resulted. Four fetuses showed no changes in polyribosome sedimentation profile or in sedimented polyribosome supported in vitro protein synthesis. Four showed changes in sedimentation profiles indicating partial breakdown of polyribosomes and significant decreases in polyribosome supported in vitro protein synthesis. Finally, 8 fetuses showed almost complete disaggregation of cerebral polyribosomes into 80-S monosomal units. This was associated with marked reduction in sedimented ribosomal mediated in vitro portein synthesis. Both the disaggregation of polyribosomes and the associated loss of in virto protein synthesis were related to the severity of the acidosis produced by asphyxia.


The dilemma of childhood optic gliomas.

The dilemma of childhood optic gliomas. A retrospective series reviewing 28 cases of optic glioma in childhood is reported from the Columbus Children's Hospital. The literature is reviewed from a clinical and pathological standpoint; in the pathological literature there is support for consideration of these lesions as a neoplastic entity, but none for designation of these gliomas as "hamartomas." An approach to the management of optic gliomas is presented. Treatment should be tailored to the individual patient; no clear therapeutic protocol appears to be supported by available data.


Histopathologic evaluation of three ultraviolet-activated composite resins on monkey pulps. The pulpal responses of three ultraviolet-activated composite resins, Nuva-Fil, Experimental UV No. 1 and Experimental UV No. 2, were tested on adult monkey teeth using silicate and zinc oxide eugenol (ZOE) as positive and negative controls. All materials were placed in Class V cavity preparations in Rhesus monkey teeth using approximately 48 anterior and 63 posterior teeth of both the maxillary and mandibular arches. A total of 111 teeth were utilized and all materials were evaluated at 3 days, 5 and 8 weeks. Following left ventricular perfusion, the teeth were prepared for microscopic evaluation using routine histological procedures. The 3-day pulpal response of all the ultraviolet-activated composites was slight with some disruption and vacuolization in the odontoblastic layer and a slight inflammatory response. At 5 weeks there was a reduction of the inflammatory response and the formation of reparative dentin was noted for all ultraviolet composites. The 8-week pulp response was slight, characterized by a minimal inflammatory response adjacent to the zone of reparative dentin. Generally, ZOE produced the mildest response while silicate produced the most severe response at the three time intervals.


Effects of bicarbonate on remineralization of enamel. Enamel surfaces were demineralized in acetic acid and remineralized in solutions that contained 1.5 mM calcium, 1.0 mM phosphate, and 0.0, 5, 15, and 25 mM NaHCO3, and 0.00 or 5.0 mM Naf at pH 7.0. NaF in CO2-free solutions increased the rates of calcium and phosphate uptake during remineralization. Bicarbonate in NaF-free solutions caused small increases of calcium and phosphate uptake. Bicarbonate with NaF in solutions synergistically increased calcium uptake, but did not affect phosphate uptake. Bicarbonate reduced fluoride uptake by about 50% during remineralization, and increased the Ca/P ratios of deposited minerals from about 1 (dicalcium phosphate) to approach and exceed the ratios of apatites.


The carbonate and fluoride in surfaces of remineralized enamel. Analyses were made of samples of surface enamel collected before and after acid demineralization and after subsequent remineralization in solutions containing calcium and phosphate and various levels of bicarbonate and fluoride ions. Demineralization caused a preferential loss of carbonates. Remineralization was increased by both fluoride and bicarbonate ions. With no bicarbonate in the calcifying solution, 0.25 to 4.0 mM NaF increased mineral formation and thereby caused a relative reduction of its carbonate content. With 0.5 mM NaF (10 ppm F-) and 5, 15, and 25 mM HCO-3 in remineralizing solutions more carbonate was deposited than in the absence of NaF. Five and 15 mM HCO-3 increased, but 25 mM HCO-3 decreased, deposit of fluoride in enamel.


Odontogenic myxoma - clinical concept and morphological studies. Our experience with the odontogenic myxoma of the mandible is presented. Although it is a benign, slow growing neoplasm, it is locally aggresive. Patients rarely complain of any symptoms produced by this tumor. Morphological studies included enzymes histochemistry, as well as light and electron microscopy. Two major types of tumor cells were identified. One resembled a mature fibroblast, while the other had the morphological and functional criteria of a so-called myxoblast. We would recommend therapy comprising wide resection and consecutive primary osteoplasty.


Odontoblast metabolism in rats deficient in vitamin D and calcium I: A histochemical survey. Different rat diets, deficient in calcium and/or vitamin D, were tested for varying experimental periods to obtain changes in serum calcium values, body weight gain and odontoblast-predentine morphology. One diet, R 25, used during a 14-day period, was found to induce lowered serum calcium values and an increased predentin width in incisor teeth. Rats fed this diet demonstrated an increase in alkaline phosphatase and ATP-splitting enzyme activity in dentinogenically active incisor odontoblasts. No other metabolic changes in these cells were demonstrated by the histochemical methods employed.


The relation of lipids to the mineral components in salivary calculi. Using lipid histochemical and microradiographic methods, the distribution of lipids and the inorganic component in different areas of salivary calculi was investigated. Two main groups of lipids were found, i.e. hydrophilic phospholipids and hydrophobic lipids. The hydrophilic phospholipids were shown to be composed mainly of phosphosphingosides and of the hydrophobic lipids cholesterol was demonstrated. Phosphosphingosides were localized in more or less lamellar zones as also in large homogeneous areas. In some cases the calculi were surrounded by a structureless non-mineralized zone exhibiting a strongly positive reaction for phosphosphingosides. Cholesterol was found in isolated band-formed areas. Phosphosphingosides and cholesterol were especially prevalent in areas showing a low or slight degree of mineralization. In areas showing a high degree of mineralization the corresponding lipids gave a weak or absent histochemical reaction. After EDTA-decalcification, however, an increase in the concentration of lipid was noticed in high mineralized areas, indicating a masking process. The influence of various factors on the presence and distribution of different lipids was discussed.


An autoradiographic study of experimental odontogenic cyst formation in the mouse. Proliferative populations of cells within the epithelial lining of developing experimental odontogenic cysts were identified by labeling with 3H-thymidine. Mouse molar teeth extracted from 10-day-old mice were transplanted subcutaneously and host animals were injected with 3H-thymidine 1 h prior to sacrifice. The cysts which developed around the crown of the transplanted teeth were prepared for autoradiographical examination. Labeling indices were high initially but decreased rapidly as the cysts enlarged. The pattern of labeling, however, bore no resemblance to the labeling of the reduced enamel epithelium from which the cysts developed. It was concluded that labeling activity reflects only the state of development and activity of the cyst and not the origin of the cyst.


Granular cell ameloblastoma: electron microscopic observations. A case of granular cell ameloblastoma was studied by light and electron microscopy. Histologically, the lesion resembled simple ameloblastoma, but differed from the latter in that it contained many cells with granular eosinophilic cytoplasm. Ultrastructurally, the cytoplasmic granules were identified as lysosomes. These organelles were particularly abundant in cells situated in the center of islands and clusters of neoplastic cells. Since no cytoplasmic components could be identified in any of these lysosomes, it was concluded that they are not engaged in autophagy or in remodeling of the neoplastic cells.


Theophylline for treatment of asthma.

Theophylline for treatment of asthma. Theophylline has become useful not just as an acute bronchodilator, but also as a major prophylactic agent for the suppression of symptoms of chronic asthma. Efficacy and toxicity both relate closely to serum concentration. Variable elimination rates for the drug among individuals require careful individualization of dosage guided by measurement of serum theophylline concentration to maintain safe and effective serum theophylline concentrations. Fixed-dose combinations with ephedrine are more likely to add toxicity than benefit compared with theophylline alone if the latter is used in an adequate dose. Choices among the vase array of products available can be further simplified by considering cost and convenience of the various formulations. When used appropriately, theophylline has been successful in the essential elimination of asthmatic symptoms in over 70% of asthmatic children seen at a tertiary care referral center.


Plasma somatomedin and growth hormone values in children with protein-calorie malnutrition. This study was undertaken to define the interrelationships of somatomedin, growth hormone, and an inhibitor of SM in protein-calorie malnutrition. Twenty-seven patients, eight to 60 months of age, were studied. Twenty-one well-nourished children acted as controls. SM was significantly depressed at Days 2 and 8 (p less than 0.01) but was not different from controls at Day 29 and 50. In ten out of 27 Day 2 samples and six out of 27 Day 8 samples an inhibitor was identified. When SM values were compared to simultaneous hGH levels, there was an inverse relationship. The low SM levels in the face of markedly elevated hGH levels suggests a functional block in the synthesis and/or release of SM.


Precocious puberty in juvenile hypothyroidism. Three children, two girls and one boy, presented with hypothyroidism and precocious sexual development. Serum TSH and PRL were elevated in all cases. In the male patient, pituitary stimulation with TRH produced exaggerated TSH and PRL responses. Elevated serum LH, FSH, estrone, estradiol, and an attenuated gonadotropin response to GnRH were documented in the male patient and in one of the female patients. Serum testosterone was elevated for age in one of the female patients but prepubertal in the male patient. After treatment with L-thyroxine, elevated hormonal levels decreased to normal in one patient who returned for follow-up. Evidence is presented that precocious puberty in juvenile hypothyroidism may be the result of PRL hypersecretion.


Production of Sarcocystis cruzi sporocysts by dogs fed experimentally infected and naturally infected beef. Twenty-one coccidia-free dogs were fed either experimentally infected or naturally infected beef in 4 experiments. The pretreatment period ranged from days 9 to 33; 7 dogs shed their first sporocysts on day 12. The full length of the patent period was not determined because most dogs were still shedding sporocysts when experiments ended on days 40 and 60. Although the patent period was long, sporocysts were not shed continuously; the total number of days sporocysts were actually shed ranged from 3 to 40. The average number of sporocysts shed by dogs fed 454 to 908 g of beef ranged from 861,000 to over 20 million. Most sporocysts were shed from days 15 to 30; peak numbers were shed on days 23 and 24. Average peak numbers ranged from 145,000 to 408,400 sporocysts. Such data can be applied to facilitate collection of sporocysts in the laboratory or to estimate the potential for transmission of infective organisms under field conditions.


Peripheral total parenteral nutrition employing a lipid emulsion (Intralipid): complications encountered in pediatric patients. The clinical records of 180 pediatric patients who received Intralipid via peripheral veins at a single institution (1964-1977) were retrospectively analyzed, with particular reference to the complications of this form of therapy. Intralipid was used in a dose range of 2--4 g/kg/day in order to supply 40% of the daily calorie requirements. The patients were neonates, infants, children, and adolescents with a wide range of clinical diagnoses. Local complications associated with Intralipid therapy were minimal. Transient elevations in serum enzyme levels (SGOT, SGPT, and LDH) were observed in 4% of patients, but all of these returned to the normal range after cessation of therapy. Ten patients had histologic evidence of cholestasis, the significance of which is discussed. The lipid emulsion was employed in patients with preexisting hyperbilirubinemia with concomitant resolution of jaundice. Intralipid was administered to patients with known severe thrombocytopenia (secondary to sepsis or myelosuppression) with return of the platelet counts to normal levels during the course of infusion therapy. The use of Intralipid in patients with established sepsis did not delay its response to conventional surgical or antibiotic therapy. There were no instances of the "overloading" syndrome observed.


Dihydroergotoxine: separation and determination of four components by high-performance liquid chromatography. The evaluation of a new high-performance liquid chromatographic method is described. It permits the separation and determination of the four components of dihydroergotoxine (dihydroergocristine, dihydroergocornine, dihydro-alpha-ergocryptine, and dihydro-beta-ergocryptine) in a single step. On reversed-phase microparticles, complete baseline separation is possible with different mobile phases containing about 10(-2) M base. The analysis of dihydroergotoxine mesylate drug substance or its dosage forms can be carried out in about 15 min. No reference substance is required for the determination of the proportions of the components. This method is simple and exhibits high accuracy, reproducibility, and selectivity. It permits the analytical control of dosage forms containing dihydroergotoxine mesylate to ensure that they comply with the specifications for the drug substance used in clinical and pharmacological studies.


Effects of intravenous cocaine, diethylpropion, d-amphetamine and perphenazine on responding maintained by food delivery and shock avoidance in rhesus monkeys. The effects of perphenazine, cocaine, diethylpropion and d-amphetamine on responding maintained by both food delivery and electric shock avoidance were determined using a multiple schedule of reinforcement in rhesus monkeys. This schedule had three components, each separated by a timeout: a fixed-ratio schedule of food delivery, a schedule of spaced responding (differential reinforcement of low rates) maintained by food delivery and a fixed-ratio schedule of shock avoidance. Control rates of responding on both ratio schedules were similar and were high relative to the low rates generated by the schedule of spaced responding. Perphenazine decreased rates on all three schedules in a dose-dependent fashion. All three psychomotor stimulants decreased food-maintained ratio responding at doses which had little effect on or increased rates of shock avoidance. Except for diethylpropion and d-amphetamine in one animal in which rates were increased, low rates of spaced responding were also decreased.


The effects of acute and chronic phencyclidine on schedule-controlled behavior in the squirrel monkey. The effects of acute and chronic administration of phencyclidine (PCP) were examined in five male squirrel monkeys trained to respond on a chain fixed-interval fixed-ratio schedule of food presentation. Acute PCP (0.01-0.60) mg/kg i.m.) produced dose-related decreases in response rate during both components of the schedule. Both components were equally affected by the drug. The effects of the drug on fixed-interval response rate were dependent on the control rate of responding in corresponding segments of the interval. After the initial dose-response determination, the subjects were placed on an individualized regimen of chronic PCP administration lasting from 82 to 126 days, beginning with daily injections for 2 days alternating with saline injections for 2 days, progressing to four injections daily. No evidence of physical dependence was seen upon withdrawal of the drug. Redetermination of the dose-response function for PCP (0.03-1.0 mg/kg i.m.) demonstrated a nearly 2-fold shift to the right of both the fixed-interval and fixed-ratio dose-response curves, indicating tolerance. In addition, the subjects' behavior recovered sooner from a dose of PCP (0.60 mg/kg i.m.) given after the chronic regimen than from the same dose given before the chronic regimen. The results demonstrate that tolerance can occur to the behavioral effects of PCP in the squirrel monkey.


Conflict behavior in the squirrel monkey: effects of chlordiazepoxide, diazepam and N-desmethyldiazepam. Dose-response profiles were determined for chlordiazepoxide, diazepam and N-desmethyldiazepam in a squirrel monkey punishment (conflict) procedure. The monkeys were trained to lever press under a food-maintained concurrent schedule consisting of an unpunished 6-minute variable interval (VI) schedule, and a 1.5-minute VI schedule, on which responses were punished intermittently (24 response variable ratio) with electric footshocks. The three benzodiazepines effectively increased responding that had been suppressed by punishment; they had inverted U-shaped dose-effect curves. The minimum effective doses for increasing punished responding were: diazepam less than or equal to 0.31 mg/kg p.o.; N-desmethyldiazepam = chlordiazepoxide = 0.62 mg/kg. As a model to assess potential antianxiety activity, this procedure possessed excellent sensitivity and reliability. The following observations were also made. 1) During initial training, as shock intensity was increased and punished responding became suppressed, some monkeys exhibited an increase in unpunished response rates. This may have represented "positive behavioral contrast," but response rate changes were associated with changes in the amount of time the monkeys allocated to each schedule. 2) At certain dose levels, all three compounds exerted antipunishment effects 24 hours after administration. 3) As was reported previously for rats, when the monkeys had no previous drug experience ("drug-naive") they were more sensitive to the depressant effects of the benzodiazepines. With repeated administration, there was a reduction in this sedation and a concomitant increase in the antipunishment effect. This phenomenon was dose- and animal-dependent.


Fluxes and distribution of calcium in rat liver cells: kinetic analysis and identification of pools. 1. Fluxes and distribution of Ca were studied in the perfused rat liver. Kinetic analysis of (45)Ca exchange revealed three compartments with time constants of 4, 14 and 223 min, pool sizes of 250, 385 and 670 mumole.kg(-1) wet wt. respectively, and one non-exchangeable compartment of 400 mumole.kg(-1).2. (45)Ca uptake by in situ mitochondria followed, as a function of cell loading time, a mono-exponential function with a time constant of 16 min. This suggests that the second compartment may be identified as the intracellular pool of Ca. The calculated cell transmembrane flux of Ca was 28 mumole.kg(-1) min(-1) or 0.17 p-mole.cm(-2).sec(-1).3. The maximum (45)Ca uptake by in situ mitochondria was 2.3 n-mole.mg(-1) of protein which represents, on the basis of 50 mg of mitochondrial protein per g of fresh liver, 115 mumole.kg(-1) or 30% of the cytoplasmic pool. A pool of 10.8 n-mole.mg(-1) protein (or 540 mumole.kg(-1)) of non-exchangeable Ca (at steady state) was probably in the form of Ca phosphate precipitated in the mitochondrial matrix.4. Extracellular Ca pools were studied using competitor of Ca binding (La) or Ca chelators (EGTA). La displaced specifically a homogeneous pool of Ca (tau = 5.1 min) which represented a fraction (55 mumole.kg(-1)) of the rapidly exchangeable Ca (first compartment) without perturbing other external pools. On the other hand, EGTA displaced completely that compartment, and about 85% of the Ca of the third compartment. These results suggest that the first and the major fraction of the third compartments are extracellular. They account for 63% of total exchangeable Ca.5. A model of distribution and exchange of Ca in hepatocytes is proposed.


Search for a natriuretic mechanism sensitive to sodium in the brain of the monkey. 1. The effects of hypertonic saline infusion into the third ventricle were investigated in ten monkeys which were pre-operated, trained, and used in the conscious state under controlled conditions. 2. In non-hydrated monkeys, intraventricular infusion of NaCl 1.0 M, 0.01 ml./min for 30 min did not affect urine volume or Na output but produced a small increase in urine osmolality. Comparable infusion of NaCl 0.15 M had no effect on any parameter. 3. In monkeys undergoing water diuresis (with i.v. infusion of 5% dextrose), intraventricular hypertonic saline produced large reciprocal changes in urine volume and osmolality while urine Na showed no significant change. The effects on urine volume and osmolality were greater than those of lysine-vasopressin 30 m-u./kg i.v. 4. The absence of natriuresis after intraventricular hypertonic saline infusion in the monkey was in notable contrast to the results reported in lower species. However, the data suggested that the infusion probably released ADH as in other species.