Macrophage-agglutinating factor produced in vitro by BCG-sensitized lymphocytes. Supernatant fluids from cultures of BCG-sensitized rabbit lymph node and spleen cells contained a factor that strongly agglutinated normal rabbit alveolar macrophages within 3 min at room temperature. In contrast, fluids from nonsensitized cell cultures did not agglutinate normal rabbit alveolar macrophages. This factor was designated macrophage-agglutinating factor (MAgF) because it is similar to the previously described factor found in lung lavages of rabbits exhibiting a BCG-induced pulmonary delayed hypersensitivity reaction. The kinetics of MAgF production in vitro by sensitized lymph node cells and its inhibition by puromycin and actinomycin D suggest active synthesis; sensitized spleen cells exhibited kinetics resembling release rather than synthesis. Studies on purified lymphocyte and macrophage populations from sensitized spleen and lymph nodes indicated that lymphocytes are responsible for MAgF production. However, MAgF production was not induced in normal cells incubated in vitro with concanavalin A or phytohemagglutinin. Fractionation of cell culture supernatant fluids in Sephadex G-100 or Ultrogel AcA-34 clearly separated MAgF from migration inhibition factor; MAgF was present in the void volume of the eluates, suggesting a molecular weight of over 400,000, whereas migration inhibition factor was recovered in the same peak as albumin. The role of MAgF in vivo is unknown, but it is postulated that it may cause the adherence of macrophages during granuloma formation.


Cryptococcus neoformans: size range of infectious particles from aerosolized soil. Although cryptococcosis is characterized as a chronic central nervous system disease, it is generally accepted that the lungs are the primary portal of entry for the etiological agent. Despite this, there is a distinct lack of evidence that viable airborne particles of Cryptococcus neoformans are small enough to reach the alveoli. Two encapsulated strains and one nonencapsulated strain of C. neoformans were inoculated into 250-g quantities of sterile soil. Throughout the 0 to 12 weeks of incubation, this soil was aerosolized in a sealed chamber with a Waring blender. Samples of the resultant dust cloud were taken with an Anderson air sampler from which the numbers and sizes of viable airborne particles were determined. Of the viable organisms aspirated into the air sampler, 15% were 0.65 to 2 micron in diameter. As incubation time in soil increased, the size of the particles decreased, and increased numbers of C. neoformans cells 0.65 to 2 micron in diameter were isolated. The presence of viable cells less than 2 micron in soil aerosols indicated that, under certain conditions in nature, C. neoformans cells exist in sizes that are capable of deep lung deposition.


In vivo studies on the cellular source of migration inhibitory factor in mice with delayed hypersensitivity. Mice sensitized intravenously with 300 microgram of BCG cell walls in Drakeol-Tween 80 and challenged intravenously 3 weeks later with 50 mg of old tuberculin released migration inhibitory factor (MIF) into the circulating blood in quantities that could be detected in serum dilutions of 1:64 to 1:128. When thymus-derived lymphocytes (T-cells) were absent at the time of sensitization, as in neonatally thymectomized mice or in athymic nude mice, detectable amounts of MIF were not formed. Sensitized mice treated with either anti-theta serum or anti-bone marrow-derived lymphocyte (B-cell) serum before intravenous challenge with old tuberculin released reduced amounts of MIF into the circulation. Mice lethally irradiated, reconstituted with B-cells, sensitized with BCG cell walls, and then challenged intravenously 3 weeks later with old tuberculin did not release MIF into the circulation. When T-cells were injected at least 10 days before challenge, however, MIF appeared.


Development of of macrophage migration inhibition in rabbits infected with virulent Treponema pallidum. Peritoneal exudate cells from rabbits infected with Treponema pallidum Nichols were used as indicators of macrophage migration inhibitory factor activity. Between 3 and 15 weeks after infection, the migration of peritoneal exudate cells was inhibited in the presence of 3 to 25 microgram of T. phagedenis biovar Reiter protein per ml. Before this period, the migration patterns of peritoneal exudate cells from infected animals were uninhibited and similar to those from noninfected control rabbits. These observations were correlated with the development of active cell-mediated immunity during experimental T. pallidum infection.


Gliding motility and actinomycin D sensitivity of Fusobacterium nucleatum and other gram-negative rods. Six strains of gram-negative anaerobic fusiform rods (Fusobacterium and Bacteroides spp.), isolated from deep subgingival locations in humans, were examined for (i) gliding motility in slide cultures, (iii) cell densities on nutrient agar surfaces, and (iii) sensitivity to actinomycin D. Known gliding (FBt) and nonmotile (NM) strains of Myxococcus xanthus served as controls for the gliding and cell density experiments. In the present study, three strains of gram-negative fusiforms from the oral cavity, identified as Fusobacterium nucleatum (1a and P-1) and Bacteroides sp. (P-14), demonstrated characteristics similar to the gliding control strain M. xanthus. The only gliding oral strain sensitive to actinomycin D was a Bacteroides sp. (P-14).


Increased resistance of iron-deficient mice to salmonella infection. Nutritional iron deficiency in mice attenuated Salmonella typhimurium infection compared with both iron-substituted littermates and normal diet control animals.


Induction of nephrocalcinosis in rabbit kidneys after long-term exposure to a streptococcal teichoic acid. New Zealand white rabbits were administered soluble lipoteichoic acid from Streptococcus pyogenes 1-RP41 on alternate days for up to 30 days. An increased incidence of renal cortico-medullary calculi was observed after day 21; the use of fluorescent-labeled anti-teichoic acid antibody located teichoic acid predominantly in the cortical-associated tubules.


Canine migration inhibitory factor: effect of Corynebacterium parvum administration. Peripheral blood lymphocytes from dogs sensitized to streptolysin O (SLO) were assayed for migration inhibitory factor (MIF) production by the indirect MIF test, using guinea pig peritoneal exudate cells as the source of macrophages. A specific direct correlation was established between the degree of inhibition of migration and the concentration of SLO-stimulated supernatants from lymphocyte cultures (SLO-S) of untreated normal dogs. Undiluted SLO-S inhibited migration by 66.8%, whereas a dilution of 1:64 elicited a 3% inhibition. In parallel tests, purified protein derivative stimulation of lymphocytes from BCG-vaccinated dogs produced 92.6% inhibition. The effect of Corynebacterium parvum on SLO-specific MIF production was evaluated in three groups of dogs administered a single intramuscular injection of C. parvum at 5 or 50 mg/m(2) or 50 mg/m(2) in suspension with 10 mg of methylprednisolone. Inhibition of migration of macrophages exposed to a 1:4 dilution of SLO-S from dogs inoculated with C. parvum (5 mg/m(2)) was 33% greater (mean inhibition, 75%) than the same SLO-S dilution from uninoculated normal dogs (mean inhibition, 42%) (P < 0.0002). Similarly, lymphocytes from dogs administered 50 mg/m(2) caused an enhancement of migration inhibition, with a mean increase of 26% over controls (P < 0.002), whereas a dose of 50 mg/m(2) with methylprednisolone produced a 16% increase in migration inhibition (P < 0.05). The administration of C. parvum resulted in a three- to fourfold increase in the SLO-S dilution, which would reduce migration by 20% (MIF titer). This increase peaked between days 20 and 30 and lasted over 50 days post-C. parvum inoculation. These findings indicate that C. parvum specifically increases MIF production by canine lymphocytes in a linear correlation with SLO concentration and suggest its use as a stimulant of canine immunity.


Immunological interrelationships between cholera toxin and the heat-labile and heat-stable enterotoxins of coliform bacteria. Cholera toxin (CT) and the heat-labile (LT) toxin of Escherichia coli are known to share antigenic properties. The present study examined the immunological relationship of CT and the LT and heat-stable (ST) toxins of E. coli, Klebsiella pneumoniae, and Enterobacter cloacae. The neutralizing capacity of equine CT antiserum and of antiserum raised in rabbits to the LT toxin of the three species of coliform bacteria was evaluated by determining their capacity to inhibit the action of purified CT and semipurified ultrafiltration preparations of the coliform LT and ST toxins in inducing water secretion as assayed by the in vivo marker perfusion technique in the rat jejunum. One milliliter of antiserum to CT and to E. coli and Klebsiella LT completely neutralized the secretory action of each of these three toxins; effective serial dilutions of CT antiserum extended to 1 to 4, whereas those of the antisera to LT were limited to 1 to 2 in most instances. One milliliter of antiserum to E. cloacae LT partially neutralized each of the three coliform LT toxins; serial dilutions were inactive. Antiserum to E. cloacae LT did not neutralize CT. Antiserum to CT and to each of the three coliform LT toxins also had a weak neutralizing effect on the ST toxins of E. coli and Klebsiella, but they did not affect E. cloacae ST. Adsorption of the antiserum to CT and to each of the three LT toxins by incubation with a heat-inactivated preparation of either the homologous or a heterologous LT toxin completely abolished the neutralizing capacity of the antisera towards both LT and ST. These observations indicate that the immunological interrelationship of CT and E. coli LT extends to the LT toxins of Klebsiella and E. cloacae and, further, that these immunological properties are shared to a lesser extent by the ST toxins of E. coli and Klebsiella.


Anticomplementary activity of Fusobacterium polymorphum in normal and C4-deficient sources of guinea pig complement. Fusobacterium polymorphum has been isolated from the gingival crevice in humans and has been implicated in the immunopathology of periodontal diseases. The presence of alternative complement pathway factors in gingival crevice material suggests the contribution of this process in the manifestation of the local host response. A complement consumption assay was used to show that the anticomplementary activity of a cell-wall preparation from F. polymorphum in guinea pig complement progressed optimally at 37 degrees C and suboptimally at 0 degrees C. Similar levels of complement consumption were recorded at both temperatures, but a higher concentration of cell walls was required to demonstrate maximum activity in the cold. Augmentation experiments with additional normal and heat-treated complement showed the presence of heat-labile factors that controlled and limited the consumption of complement to a constant proportion of the total amount present. These same experiments indicated that there were no reactive antibodies in the complement used in the assay. Similar tests with C4-deficient guinea pig sera confirmed that F. polymorphum cell walls were capable of generating alternative complement pathway activity in guinea pig sera.


Prolonged survival of virulent Treponema pallidum (Nichols strain) in cell-free and tissue culture systems. Survival of Treponema pallidum was found to be prolonged in the presence of tissue culture. Of the 12 cultures studied, cottontail rabbit epithelium (Sf1Ep) supported T. pallidum for the longest time. In horizontal Leighton tubes with reduced medium and an atmosphere of 5% CO2 in N2, the 50% survival time (ST50) was 5 to 6 days for treponemes associated with monolayers of Sf1Ep cells. Comparable cell-free tubes had ST50 values of less than 4 days. In vertical Leighton tubes containing 6 ml of prereduced medium incubated aerobically, gradients of O2 tension and redox potential were established. Attachment and survival of T. pallidum were greatest at a depth of about 10 to 20 mm. Motility was between 70 and 95% in this area throughout the first 14 days of incubation. Occasionally, greater than 50% motility was observed for as long as 21 days. The redox potential and O2 tension in the optimal area of gradient cultures were reproduced by adjusting the medium depth in a shell vial culture system containing cells on a horizontal cover slip. Treponemes associated with the cell monolayer in both gradient and shell vial cultures were still virulent after 21 days in vitro. The dilution of testis extract and the concentration of T. pallidum were found to be important factors in survival of T. pallidum.


Role of bacterial phospholipases in serum-mediated killing of Escherichia coli. The importance of bacterial phospholipases during serum-mediated killing of Escherichia coli was examined by using wild-type DR+ DS+ and an isogenic phospholipase-deficient mutant DR- DS-. No difference in serum sensitivity was observed when the parental DR+ DS+ and mutant DR- DS- strains were exposed to various concentrations of normal guinea pig serum. Examination of the free fatty acid (FFA) and lipid composition during serum-mediated killing of the two E. coli strains indicated that FFA release occurred only in the parental DR+ DS+ strain. No FFA release or lipid degradation was detected in the mutant DR- DS- strain during serum killing. The addition of heat-inactivated E. coli antiserum (rabbit) to normal guinea pig serum caused FFA release in both E. coli strains. This FFA release was found to be independent of serum-mediated killing and due to a highly active and heat-resistant rabbit serum phospholipase that hydrolyzed the bacterial lipids after serum killing. The data presented indicate that serum-mediated killing of E. coli is independent of FFA release and that activation of bacterial phospholipases and the resulting release of FFA are only a result rather than a cause of serum-mediated cell death.


Extracellular neuraminidase production by group B streptococci. Neuraminidase (sialidase) activity in concentrated culture filtrates of group B streptococci was measured with bovine submaxillary mucin as substrate. Group B streptococcal neuraminidase was not active on human alpha-1 acid glycoprotein and did not show increased activity on bovine submaxillary mucin that had been O-deacetylated by alkaline treatment. The enzyme was produced in a variety of media, including a chemically defined medium (FMC; Terleckyj et al., Infect. Immun. 11:649-655, 1975) supplemented with bovine serum albumin or human serum albumin. Maximal levels of activity were present in filtrates from cells grown in a dialyzable fraction of Todd-Hewitt broth harvested during the late exponential phase of growth. Dramatic decreases were seen when filtrates from the late stationary phase were assayed. The decrease in specific activity during the stationary phase was shown to be due to proteolytic digestion of neuraminidase and not to the elaboration of an extracellular neuraminic acid aldolase.


Cell-mediated immunity to Nocardia asteroides induced by its ribonucleic acid protein fraction. Guinea pigs immunized with ribonucleic acid protein fraction from Nocardia asteroides developed high cell-mediated immunity within 2 weeks, as shown by increased macrophage microbicidal activity and macrophage migration inhibition. This immunity protected against intravenous and subcutaneous challenge with N. asteroides and persisted for at least 70 days.


Interactions between salmonellae and macrophages of guinea pigs. IV. Relationship between migration inhibition and antibacterial action of macrophages. The in vitro macrophage migration inhibition test was used to detect the development of delayed-type hypersensitivity in guinea pigs infected with Salmonella typhimurium. Four different preparations from supernatants of S. typhimurium cultures were used as the antigens in this test. They included the concentrated bacterial antigens, the high-molecular-weight (>50,000) antigens, the ammonium sulfate-precipitated antigens, and the ribonuclease-treated antigens. All four antigen preparations were shown to inhibit the migration of peritoneal macrophages of salmonella-infected (immune) guinea pigs from capillary tubes, in comparison with cells of normal control animals. By use of the high-molecular-weight antigens and the ammonium sulfate-precipitated antigens, the production of the migration inhibition factor(s) was elicited from cultures of lymphocytes obtained from the peripheral blood of immune guinea pigs. The activity of the migration inhibition factor(s) was demonstrated by its ability to inhibit the migration of peritoneal macrophages of normal guinea pigs from capillary tubes. In contrast, normal peritoneal macrophages exposed to products of antigen-stimulated immune lymphocytes did not exhibit an enhanced phagocytic or bactericidal action against virulent S. typhimurium as compared with those of the normal control. The present study indicated that the bacterial antigens responsible for the elicitation of the production of the migration inhibition factor from lymphocytes of immune guinea pigs are inactivated by proteolytic enzymes, but not by ribonuclease, and have molecular weights of >50,000.


Transduodenal sphincteroplasty.

Transduodenal sphincteroplasty. Sphincteroplasty is an inferior partial sphincterotomy followed by suture approximation of both the duodenal and common bile duct walls. We have described our operative technic employed in 770 sphincteroplasties performed in the past 14 years.


Transabdominal esophageal transection by using a suture device in cases of esophageal varices. We have devised a method of esophageal transection using a suture device. This method interrupts the esophageal varices directly. We have performed this operation in 30 cases with satisfactory results. Blood loss is minimal. The operation is both shorter and simpler than conventional technics.


Microsurgical renal transplantation techniques in small animals. The laboratory rat has been widely used as a preclinical model of renal transplantation. Surgical anastomosis of the small lumenal structures encountered requires strict adherence to basic principles of microsurgical technique. Nonfatal technical errors may adversely affect renal allograft function and influence experimental results. Experience obtained from a series of more than 600 rat renal transplants is discussed.


A dye-dilution method for the determination of blood flow in Cimino-Brescia arteriovenous fistulae. In 15 patients with large Cimino-Brescia fistulae, arm blood flow and cardiac output were determined by a dye-dilution technique. The mean blood flow through the arm with the shunt open was 1600 ml per min. On compression of the shunt the appearance time increased by 10 sec and there was a moderate decrease in cardiac output.


Ultrastructure of the gut neurotensin cell. In mammals, neurotensin cells occur scattered in the epithelium of the jejunum-ileum. In chicken, neurotensin cells are abundant in the region of the gizzard-duodenal junction (antrum) where they occur intermingled with numerous somatostatin and gastrin cells. The neurotensin cells in chicken, dog and man were identified at the electron microscopic level by immunocytochemistry, using the consecutive semithin/ultrathin section technique. They contain numerous electron dense cytoplasmic granules, pre-dominantly in the basal portion of the cell. It was shown that these granules are the storage site for neurotensin. The neurotensin granules are round, highly electron dense and of about the same size in the different species examined (mean diameter 260--290 nm). In dog and man the granules have a tightly applied surrounding membrane while in the chicken a relatively electron lucent zone separates the electron dense core from the granule membrane. The ultrastructure of the neurotensin granules in chicken is somewhat reminiscent of that of the gastrin granules. The mean diameter of the gastrin granules in chicken antrum is 230 nm; for the somatostatin granules the mean diameter is 305 nm.


The influence of fixation on immunoperoxidase staining of plasmacells in paraffin sections of intestinal biopsy specimens. The influence of different fixation methods on the results of immunoperoxydase staining of immunoglobulin and gastrin producing cells in gastric and duodenal mucosa was investigated. An indirect method was used on paraffin sections. It appeared that that fixatives containing sublimate gave the most consistent results, a sublimate-formaldehyde mixture being the best.


Colloidal iron binding to the surface of HeLa cells, spreading in monolayer culture. We have used the colloidal iron (CI) binding technique, adapted for transmission electron microscopy, for semiquantitative evaluation of the negative charge density at the surface of HeLa cells in monolayer culture. The surface area increases when HeLa cells spread on the substrate. This increase brings about a decrease in the thickness of the CI rim, indicating a decrease in negative surface charge density. This phenomenon implicates lowering of the electrostatic repulsion, and explains the formation of intercellular contacts at the level of spread parts of the cell. Because of lack of penetration, CI particles are absent in regions of cose apposition between cells and between cells and substrates. Absence of CI binding in broader intercellular or cell-substrate spaces was explained through masking of the anionic groups.


The localization of proteoglycans and glycoproteins in the hyaline cartilage. Antibodies to proteoglycan (PG) and glycoprotein of bovine nasal cartilage were conjugated with fluorescein isothiocyanate and with horseradish peroxidase. Hyaluronidase digestion of cartilage tissue-specimens increased the intensity of immune reactions; pronase digestion or extraction with 4 M guanidinium chloride abolished the staining. In the intercellular matrix fine filaments beaded with small granules were seen forming an irregular network. The interstices of the network are filled with collagen fibers linked together by the filaments and granules. In view of the linear conformation of core proteins of PGs and the globular conformation of glycoproteins (link proteins), it may be supposed that the granules and filaments represent these two protein components of PG-aggregates. In chondrocytes a homogeneous staining was recorded in the endoplasmic reticulum, in the juxtanuclear areas and in several smooth-walled vesicles and elongated areas situating subjacent to the cell membrane. In contrast to the extracellular immune reactions, this homogeneous intracellular staining was never enhanced by hyaluronidase digestion. This is interpreted in the sense that conformation changes of molecules secreted, and the aggregation of PGs, occur extracellularly.


[The diagnostic and therapeutic possibilities of endolaryngeal microsurgery (author's transl)]. The technique and types of anesthesia for endolaryngeal microsurgery are described, utilizing a personal series of 1283 procedures which were performed over a preceding ten-year period. The great diagnostic value of this method of examination for the early diagnosis of glottic carcinoma is shown. 54.4% of all endolaryngeal microsurgical operations performed were used for the diagnosis, therapy and control of tumors. The remaining 45.6% of cases were undertaken to eliminate benign voice=disturbing processes. The indications for surgery are discussed, with note given to the low complication rate associated with such surgery.


Integration of a behavior modification program into a traditionally oriented residential treatment center for children. The historical background and early contributions of the Emma Pendleton Bradley Hospital, the first psychiatric hospital for children in the United States, are described. The focus of this report is on the incorporation of the treatment modality of behavior modification into this traditional psychoanalytically oriented program. Beginning with exploratory studies in the mid-1960s, in recent years within the residential center there has been a separate Autistic Unit, in which the major treatment strategy is consistent application of behavior modification principles and programs. Changes that have occurred in the course of developing the behavior modification program are discussed to show some of the positive and negative aspects of introducing such a radical change into a traditional setting. Case studies are included to reveal both the failures and the successes that have been encountered in treating psychotic children with either psychotherapy or behavior therapy. Description of the current status of the program shows that treatment is comprehensive, including psychodynamic and behavioral approaches, medication when warranted, and special education. The role of parents in the program is now very different from that ascribed to them in traditional psychoanalytically guided treatment.


Role of ATP in removal of psoralen cross-links from DNA of Escherichia coli permeabilized by treatment with toluene. Removal of interstrand cross-linked from DNA was examined in Escherichia coli permeabilized by treatment with toluene. Under these conditions, the reaction requires ATP and Mg2+, and the mechanism appears to be similar to that occurring in whole cells. Under optimum conditions, the rate constant was 0.06 min-1. Genetical, physical, and biochemical analysis of the repair process suggest the following mechanism. In an ATP-dependent reaction, the uvrA and uvrB gene products cleave a phosphodiester bond on the 5' side of one arm of the cross-link, producing a 3'-OH terminus. Subsequently, DNA polymerase I (5'-3' exonuclease activity) makes a second strand cut on the 3' side of the cross-link in the same DNA strand, completing removal of the covalent link between complementary strands. The second reaction did not occur in a uvrD- strain, which had normal levels of DNA polymerizing activity. The uvrD gene may regulate the specificity or activity of the 5'-3' exonuclease of DNA polymerase I in vivo.


Role of the 2-amino group of deoxyguanosine in sequence recognition by EcoRI restriction and modification enzymes. The dG residues within the EcoRI recognition sequence of ColE1 DNA have been selectively replaced with dI. Methylation of the altered sequence by the EcoRI modification enzyme is extremely slow as compared with methyl transfer to the natural recognition site. Since the affinity of the modification enzyme for the dI-containing sequence is considerably less than that for the natural sequence, we have concluded that the 2-amino group of dG has an important role in DNA site recognition by this enzyme. In contrast, the altered site is subject to cleavage by EcoRI endonuclease at rates essentially identical with those observed with the natural sequence. These results strongly suggest that the two enzymes utilize different contacts within the EcoRI site and are consisted with our conclusion (Rubin, R. A., and Modrich, P. (1977) J. Biol. Chem. 252, 7265-7272) that the two proteins interact with their common recognition sequence in different ways.


Identification and organization of ribosomal protein genes of Escherichia coli carried by lambdafus2 transducing phage. We describe the isolation of lambdafus2, which carries bacterial DNA from the str-spc region of the Escherichia coli chromosome. Genes for 27 ribosomal proteins were found on the genome of this transducing phage by identifying the ribosomal proteins whose synthesis was stimulated after infection of UV-irradiated bacteria. These were genes for S3, S4, S5, S7, S8, S10, S11, S12, S13, S14, S17, S19, L2, L3, L4, L5, L6, L14, L15, L16, L17, L18, L22 L23, L24, L29, and L30. Subsets of these genes were identified on the genomes of the phages lambdaspc1, lambdaspc2-delta 9, and lambdaspc2-delta16, all of which carry parts of the bacterial DNA present on lambdafus2. From the known structures of these phage genomes, it has been possible to determine the relative order of many of these genes on the lambdafus2 genome and thus on the E. coli chromosome. Our evidence also suggests that the genes for S3, S17, S19, L2, L4, L16, L22, L23, and L29 are part of a single transcription unit. These results, along with the observations described in the previous and accompanying papers, indicate that the ribosomal protein genes on lambdafus2 are organized into at least four transcription units.


Organization of ribosomal protein genes of Escherichia coli as analyzed by polar insertion mutations. Several mutants of lambdaspc1 and lambdafus3 have been isolated carrying DNA insertion elements that were selected for their ability to reduce the expression of the spc gene. The sizes and locations of the insertions on the phage genomes were determined by heteroduplex analysis. They were found to be located at different positions in the Spc transcription unit. The effect of these insertions on the expression of the ribosomal protein genes carried by these phages in ultraviolet light-irradiated bacteria was investigated. The insertions at intermediate positions in the transcription unit reduced the expression of some of the genes in the unit but not others. Assuming that the genes whose expressions were reduced are distal to the insertion, it was possible to determine the relative position of most of the genes in the unit. The results indicate the order of genes in the Spc transcription unit is: promoter, L14, L24, L5, S14, S8, L6, L18, (S5, L15, L30).


Insertion of DNA carrying ribosomal protein genes of Escherichia coli into Charon vector phages. DNA fragments from lambdaspc1 and lambdafus2, carrying ribosomal protein genes from Escherichia coli, were inserted into lambda phage vectors Charon 3 and Charon 4. Eight of the resulting clones were characterized by agarose gel electrophoresis of EcoRI digests, analytical CsCl equilibrium centrifugation, and electron micrographic analysis of heteroduplexes. In each case, the identity, order, and orientation of each cloned fragment was determined. In all, 8 of the 12 EcoRI fragments of lambdafus2 were cloned in various arrangements. In the accompanying paper, genes for 15 ribosomal and related proteins and three bacterial promoters were detected in these phages. In addition, four of the hybrid phages carried fragments of lambda-DNA including the phage origin of replication (ori), the late promoter, PR', and the cohesive ends (cos site) in both orientations. The latter phages yield a circularly permuted collection of DNA molecules.


Interaction of Salmonella typhimurium with phospholipid vesicles. Incorporation of exogenous lipids into intact cells. Incubation of intact cells of Salmonella typhimurium with bilayer phospholipid vesicles results in significant transfer of vesicle lipids to the cells. The transfer requires Ca2+ or spermine, and is dependent on time, temperature, the concentration and composition of the vesicles, and the nature of the cellular lipopolysaccharide. The process results in bulk transfer of vesicle lipids to the cells rather than reciprocal molecular exchange between vesicles and the outer membrane. All components of mixed lipid vesicles, including cholesteryl oleate and lipopolysaccharide, are transferred to the cells in a ratio similar to that of the donor vesicles. The properties of the transfer process are consistent with direct fusion of vesicles with the outer membrane of the cell.


Translocation of phospholipids between the outer and inner membranes of Salmonella typhimurium. The reversibility and specificity of phospholipid translocation between the inner and outer membrane of Salmonella typhimurium has been investigated by incorporating exogenous lipids from phospholipid vesicles into the outer membrane of intact cells. Translocation of newly incorporated phospholipids to the inner membrane was demonstrated by decarboxylation of vesicle-derived phosphatidylserine and by recovery of vesicle constituents in both inner and outer membrane fractions. All Salmonella phospholipids tested, as well as phosphatidylcholine and cholesteryl oleate were effectively translocated to the inner membrane. However, no translocation of vesicle-derived lipopolysaccharide or an incomplete biosynthetic precursor of lipid A could be detected. Translocation of phospholipids and cholesteryl ester was rapid and extensive, and appeared to lead to equilibration of the lipids between the two membranes. The mechanism of intermembrane translocation has not been established, but the results are suggestive of diffusional flow across zones of adhesion between the inner and outer membranes.


Muscle transposition for treatment and prevention of chronic post-traumatic osteomyelitis of the tibia. There is considerable difference of opinion about how patients with open tibial fractures and considerable loss of skin should be treated. The incidence of long-term complications from such lesions--that is, chronic ulceration and chronic osteomyelitis--apparently is not recorded, but many cases undoubtedly exist and are under treatment by methods that are short of curative. Successful results in forty-three patients were obtained by muscle transposition and delayed skin-grafting. Early coverage of open fractures of the tibia with soft tissue will prevent the later development of osteomyelitis, ulceration, and, perhaps to some extent, non-union.


Polymeric debris in synovium after total joint replacement: histogical identification. In studying a series of synovial biopsy specimens from patients with loose joint replacements, we were able to differentiate reliably between polymethylmethacrylate and ultra high-molecular-weight polyethylene by observing alterations in their birefringence with changes in temperature. The glass transition temperature of polymethylmethacrylate (when it softens from a glass-like to a rubber-like state) is about 105 degrees centigrade. Polyethylene melts at 135 degrees centigrade. The birefringence in polyethylene returns after it has been melted and then cooled; this is not true of methylmethacrylate fragments heated above their glass transition temperature. When cooled, methacrylate fails to regain its birefringence. In addition, free particles of the two plastics were studied in vitro to validate this method of differentiation. We recommend its use routinely in histological studies of patients with loosening.


Polarized bundles of actin filaments within microvilli of fertilized sea urchin eggs. We report on the internal ultrastructure of long, finger-like microvilli which cover the surface of the fertilized sea urchin egg. Eggs were attached to polylysine-coated surfaces; their upper portions were sheared away with a stream of buffer which left behind only their plasma membranes and adjacent cytoplasmic structures. Scanning electron microscopy (EM) of such fragments revealed intact thin protoplasmic projections radiating away from the body of the cortex. By transmission EM of cortices similarly prepared on grids, small bundles of microfilaments appear as cores within the thin cytoplasmic projections. These microfilaments are shown to be composed of actin by their ability to interact with muscle heavy meromyosin (HMM). HMM-decorated microfilaments possess repeating arrowheads which uniformly point toward the cell interior. Actin bundles in the microvilli of sea urchin eggs may mediate microvillus support and elongation.


The effect of D-ser (TBU)6-LH-RH-EA10 upon gonadotropin release in normally cyclic women. D-Ser (TBU)6-LH-RH-EA10 (HOE 766) was found to be approximately 40-times as effective as LH-RH in stimulating LH and FSH release in normally cyclic women. The most pronounced reaction was seen in the late follicular phase, the least one in the early follicular phase. Maximal serum gonadotropin levels were found 4 h after the injection of HOE 766. When 5 microgram was injected every 8 h for 3 days, the initially high release of LH and FSH declined progressively to almost nil.


Analysis of biological and immunological activities in the two pools of LH released during constant infusion of luteinizing hormone-releasing hormone (LHRH) in men. In response to constant infusion of a submaximal dose of LHRH (0.2 microgram/min for 4 hours), luteinizing hormone (LH) was released in a biphasic pattern in 6 normal men. When the in vitro biologic activity of plasma LH was compared to the immunologic activity throughout the response to LHRH, the B/I ratio remained unchanged (3.6 +/- 0.5, mean +/- SE). Thus, in men the biologic activity of stored LH (acutely releasable) is not different from the presumably newly synthesized hormone that is released as the second pool during prolonged LHRH infusion.


Methyl mercaptan and dimethyl disulfide production from methionine by Proteus species detected by head-space gas-liquid chromatography. Head-space gas-liquid chromatography and mass spectrometry were used to detect and identify products formed by Proteus vulgaris, P. mirabilis, P. morganii, and P. rettgeri from a defined medium supplemented with either phenylalanine, methionine, valine, leucine, histidine, lysine, ornithine, threonine, asparagine, aspartic acid, or tryptophan. In a detailed study of the products formed by 68 strains of Proteus spp. from L-methionine, the production of large amounts of both dimethyl disulfide and methyl mercaptan was found to be a characteristic of the genus. Both sulfur products appeared within a few hours of inoculation. Dimethyl disulfide was a more sensitive indicator of growth than the spectrometric determination of optical density. This suggests that it could be useful for the rapid, automated detection of any species of Proteus.


Development of specific tests for rapid detection of Escherichia coli and all species of Proteus in urine. Proteus mirabilis was distinguishable from Escherichia coli and from several other species that may be associated with urinary tract infections when grown in a nutrient medium supplemented with 0.1 M L-methionine by the formation of large amounts of dimethyl disulfide and methyl mercaptan, which were detected by head-space gas-liquid chromatography (HS-GLC). E. coli could be detected by the same HS-GLC technique by ethanol production from methionine peptone water enriched with 1% either lactose or arabinose but not by any product from 10 amino acids tested. Ethanol from lactose was detected early in the exponential phase of growth. Significant numbers, 10(5) or more per ml, of E. coli in urine could be detected in about 5 h by ethanol production from an unshaken culture of urine in lactose methionine peptone water buffered at pH 7.2 (urine test medium); only a trace of dimethyl disulfide was produced. Significant numbers of P. mirabilis in urine could be detected in 4 h by dimethyl disulfide production and in 5 h by methyl mercaptan production from a shaken culture of urine in urine test medium; no ethanol was produced. Incubation of urine specimens in the test medium followed by examination by HS-GLC is proposed as a rapid method of detecting whether or not the urine contains significant numbers of E. coli or a species of Proteus.


Assessment of technique for rapid detection of Escherichia coli and Proteus species in urine by head-space gas-liquid chromatography. A test depending on the production of ethanol by Escherichia coli from lactose and dimethyl disulfide by Proteus spp. from methionine in the early exponential phase of growth and the detection of these products by head-space gas-liquid chromatography has been applied to 75 specimens of urine selected to provide the most stringent trial of the test. The test was found to be rapid and reliable for the commonest findings in the microbiological examination of urine. In 3 to 4 h it detected "significant" numbers (greater than 10(5)/ml) of E. coli or of Proteus mirabilis or P. inconstans A, identified as Proteus spp., in 23 urines. It recorded the absence of infection from 32 urines containing borderline or "not significant" numbers of any organism. Significant numbers of other organisms in 13 urines were not mistaken for E. coli or Proteus spp. However, the test was less successful for some less common findings. Klebsiella ozenae in significant numbers in one urine was mistaken for E. coli. P. morganii in significant numbers in one urine was not detected. E. coli or P. mirabilis mixed with significant numbers of another organism were not detected in four out of five urines. The technique is simple and could be automated. It appears to merit more extensive trial in a hospital laboratory and further development to detect and correctly identify more species that cause urinary tract infections.


Survival of bacteria from human dental plaque under various transport conditions. The effects of transport media, temperature, and anaerobiosis on the survival of bacteria from human supragingival dental plaque were studied. Individual samples were obtained by passing sterile, unwaxed dental floss through the interproximal spaces. The plaque-bearing portion of floss was immediately placed in vials containing reduced transport fluid, viability-preserving microbistatic medium, or reduced salt solution transport fluid. Plaque samples were dispersed by ultrasonic oscillation, serially diluted, and plated in duplicate on MM10-sucrose-blood agar, mitis salivarius bacitracin agar, and Rogosa tomato juice agar. Initial viable counts (time 0) were compared with viable count determinations after 48- and 72-h storage. Quantitative recovery (>30%) of various groups of oral bacteria was accomplished from both reduced transport fluid and viability-preserving microbistatic medium after 48- and 72-h storage. Storage of dental plaque in reduced salt solution proved unsatisfactory for most bacteria (less than 10% survival). Since growth of some bacteria may occur in viability-preserving microbistatic medium and the charcoal present interferes with colonly enumeration on low-dilution plates, we found reduced transport fluid to be the most suitable medium for transport and recovery of bacteria from supragingival dental plaque. Subzero storage (-196 and -40 degrees C) did not enhance the survival of bacteria from dental plaque; storage at moderate (5 and 20 degrees C) temperatures gave better recovery of viable bacteria. Survival after anaerobic or aerobic storage was comparable for total colony-forming units; however, anaerobic storage enhanced survival of Streptococcus mutans and Lactobacillus. Since these organisms are specifically associated with dental caries, anaerobic techniques are preferred for caries activity testing of plaque.


Evaluation of candida precipitin and agglutinin tests for the diagnosis of systemic candidiasis in burn patients. Candida agglutinin and precipitin tests were performed on a group of burned patients without evidence of systemic candidiasis to determine whether tests could be used to distinguish between colonization and systemic disease, as has recently been reported. There were considerable numbers of false-positive results using both tests. These false-positive results occurred regardless of the candida colonization status of the patient. We conclude that these tests are of little diagnostic value in burn patients.


New blood culture medium.

New blood culture medium. A growth medium with a specific oxidation-reduction potential containing peptone, dextrose, sodium succinate, sodium lactate, gelatin, sodium bicarbonate and blue tetrazolium, an indicator dye, in a tris(hydroxymethyl)aminomethane buffer was used to detect the presence of microorganisms in blood. The procedure involved the introduction of blood (and bacteria) into the growth medium with the dye in its colorless state. As the bacteria grew, they converted the dye to a visible blue color (formazan) with their reductases. The growth medium served as its own contamination control, since microbial growth and be detected by a color change before it was used for blood culture. The experiments described herein demonstrate that the composition of this medium (with the dye) provides a unique system that is able to make a reliable and rapid detection of both gram-positive and gram-negative microorganisms and yeasts (Candida albicans) commonly associated with bacteremia.


Rapid slide coagglutination test for identifying and typing group B streptococci. A Cowan I strain of Staphylococcus aureus was labeled with either group B streptococcal grouping or typing antiserum. These antibody-labeled reagent cells (ARC) were used in a slide coagglutination test to identify and type group B streptococci from blood agar plates. All streptococci were also identified by the standard Lancefield capillary precipitin test. In a blind study, all 141 group B streptococci were correctly identified by the coagglutination grouping test. None of the 148 non-group B streptococci caused agglutination of ARC. The coagglutination grouping test required an acid extract prepared from only four colonies and could be completed less than 30 min after colonies were removed from plates. The coagglutination typing test correctly identified 98.6% of the types of the 141 group B streptococcal strains tested. At least 88.6% of these streptococci could be typed directly from blood agar plates within 5 min by the coagglutination typing test. The remaining 11.4% of the group B streptococci were acid extracted (less than a 30-min procedure), and the extract was used for coagglutination typing. Coagglutination typing can be performed with only four colonies. The coagglutination grouping and typing tests are inexpensive, rapid, reliable, and easy to perform.


Detection of bacteriuria by automated electrical impedance monitoring in a clinical microbiology laboratory. An apparatus capable of rapidly detecting changes in electrical impedance was utilized for the continuous monitoring of bacterial growth in routine urine specimens in a clinical laboratory. In a trial study, 200 clinical specimens analyzed by the electrical impedance method resulted in an average detection time of 2.5 h for 41 clinically significant specimens, whereas conventional methods for bacterial isolation required overnight culture. Those specimens positive by the electrical impedance monitoring but negative by conventional bacteriological methods accounted for less than 2% of the total number of positive specimens, whereas electrical impedance-negative but conventional culture-positive specimens accounted for ca. 4%. Electrical impedance apparatus in clinical microbiology laboratories could provide rapid screening of clinical urine specimens as well as accurate detection of bacterial growth.


Superoxide dismutase in Bacteroides fragilis and related Bacteroides species. Superoxide dismutase (SOD) activity was demonstrated in cell-free extracts of Bacteroides fragilis, Bacteroides vulgatus, Bacteroides distasonis, Bacteroides ovatus, and Bacteroides thetaiotaomicron. The strains were grown under anaerobic conditions in Trypticase soy broth, and the specific activity of SOD in the extracts was, in most strains, higher than in cell-free extracts of Escherichia coli B grown under anaerobic conditions. Isoelectric focusing of the extracts in polyacrylamide gel demonstrated distinct forms of SOD in the different species.


Rapid deoxyribonuclease test with methyl green. A 4-h deoxyribonuclease test using methyl green to differentiate Serratia from other Enterobacteriaceae was developed. The tests agreed 100% with an overnight plate test for 100 Serratia, 83 Enterobacter, and 6 Klebsiella species.


Rapid screening method for enterotoxigenic Escherichia coli. A rapid screening method for detection of Escherichia coli producing heat-labile enterotoxin is described. Single colonies are transferred directly from primary culture plates into 96-well microculture plates containing 0.3 ml of brain heart infusion broth in each well. After 24 h at 37 degrees C, each brain heart infusion broth culture is assayed by the miniculture method in the corresponding well of a microculture plate in which Y-1 mouse adrenal tumor cells have been grown. All enterotoxigenic isolates detected by this method were confirmed in the assay but with culture supernatants.


In vitro cell-mediated cytotoxicity in primary biliary cirrhosis and chronic hepatitis. Dysfunction of spontaneous cell-mediated cytotoxicity in primary biliary cirrhosis. The in vitro cytotoxic function and target cell specificity of peripheral blood lymphocytes from selected patients with primary biliary cirrhosis and hepatitis B surface antigen-negative chronic hepatitis were investigated using 51Cr-labeled human Chang and EL-4 mouse sarcoma cell targets in assays of spontaneous cell-mediated cytotoxicity (SCMC) and mitogen-induced cellular cytotoxicity (MICC). In addition, antibody-dependent cellular cytotoxicity (ADCC) against Chang cells was assessed. At an effector-to-target cell ration of 100:1, the mean SCMC against Chang cells was much less in patients with primary biliary cirrhosis than that in either the controls (P less than 0.001) or the patients with chronic hepatitis (P less than 0.005) whereas the value for patients with chronic hepatitis did not differ significantly from that of the controls. The mean SCMC against EL-4 mouse sarcoma cells was also less in patients with primary biliary cirrhosis than in controls (P less than 0.005) whereas the value for chronic hepatitis was not significantly different from that of the controls or patients with primary biliary cirrhosis. In contrast, MICC against both targets and ADCC against Chang cells were similar for each group. Comparison of SCMC and MICC against both target cells, measured simultaneously, showed similar cytotoxic potenital against both target cells for each group. Effector cells capable of mediating cytotoxicity in each assay were defined by testing the cytotoxic function of lymphocyte subpopulations isolated from two representative patients with each disease using techniques of immunoabsorbent affinity chromatography and Fc receptor binding to antigen-antibody complexes. In both primary biliary cirrhosis and chronic hepatitis SCMC and ADCC were mediated by a subpopulation of lymphocytes which lack surface immunoglobulin (sIg-) and bear Fc receptors (Fc+). In contrast, MICC was mediated by sIg- cells which lack Fc receptors. Lymphocytes bearing sIg- were not cytotoxic in any assay. These results establish a difference in cytotoxic function in primary biliary cirrhosis and chronic hepatitis by defining the presence of a defect in spontaneous cytotoxic function of sIg-, Fc+ lymphocytes against Chang cells in primary biliary cirrhosis.


Somatostatin- and epinephrine-induced modifications of 45Ca++ fluxes and insulin release in rat pancreatic islets maintained in tissue culture. The effects of somatostatin and epinephrine have been studied with regard to glucose-induced insulin release and (45)Ca(++) uptake by rat pancreatic islets after 2 days in tissue culture and with regard to (45)Ca(++) efflux from islets loaded with the radio-isotope during the 2 days of culture. (45)Ca(++) uptake, measured simultaneously with insulin release, was linear with time for 5 min. (45)Ca(++) efflux and insulin release were also measured simultaneously from perifused islets. Glucose (16.7 mM) markedly stimulated insulin release and (45)Ca(++) uptake. Somatostatin inhibited the stimulation of insulin release by glucose in a concentration-related manner (1-1,000 ng/ml) but was without effect on the glucose-induced stimulation of (45)Ca(++) uptake. Similarly, under perifusion conditions, both phases of insulin release were inhibited by somatostatin while no effect was observed on the pattern of (45)Ca(++) efflux after glucose.Epinephrine, in contrast to somatostatin, caused a concentration-dependent inhibition of the stimulation of both insulin release and (45)Ca(++) uptake by glucose. Both phases of insulin release were inhibited by epinephrine and marked inhibition could be observed with no change in the characteristic glucose-evoked pattern of (45)Ca(++) efflux (e.g., with 10 nM epinephrine). The inhibitory effect of epinephrine on (45)Ca(++) uptake and insulin release appeared to be mediated via an alpha-adrenergic mechanism, since is was abolished in the presence of phentolamine. Somatostatin inhibits insulin release without any detectable effect upon the handling of calcium by the islets. In contrast, inhibition of insulin release by epinephrine is accompanied by a partial inhibition of glucose-induced Ca(++) uptake.


Biosynthesis of the third component of complement (C3) in vitro by monocytes from both normal and homozygous C3-deficient humans. Human monocytes synthesized the third component of complement (C3) up to 5 wk in vitro. Evidence for net C3 synthesis was based on (a) incorporation of 14C-labeled amino acids into C3 protein, (b) indentity of the allotype of C3 produced in vitro with that of the doner's serum C3, even in the presence of carrier C3 protein of a different allotype; (c) correspondence of electrophoretic mobility, size, and subunit structure of C3 protein produced in vitro with serum C3; (d) inhibition of C3 production with cycloheximide. Monocytes from two unrelated C3-deficient patients were studied under conditions that supported C3 synthesis by normal monocytes. Serum from each of the patients contained less than 1% of the normal C3 concentration, buth their monocytes produced C3 at approximately equal to 25% of the normal rate when studied after 2 wk in vitro. The C3 produced in vitro by monocytes from one of the patients had the molecular weight of normal serum C3 and dissociated appropriately under reducing conditions. Monocytes from C3-deficient patients could not be distinguished from normals on the basis of morphology, rosetting with C3-coated erythrocytes, or rates of C2, and total protein synthesis.


Apparent cytotoxicity of base metal casting alloys. The cytotoxic potentials of three Ni-Cr based alloys and an Fe-Cr based alloy were assessed. In tissue culture, casting of all four test materials and powders of the Fe-Cr based material did not elicit adverse cellular changes. However, cultures containing Ni-Cr powders showed prominent zones of lysis and cell alteration. The cytotoxic potential of products obtained from the surfaces of casting alloys may offer an expedient means for estimation of long-term clinical toxicity.


Iodinated E. coli 70S ribosomes as a radiocolloid of uniform particle size for lymph-node and liver scanning. As an example of the use of biologic particles as carriers for radioactive tracers, E. coli 70S ribosomes were labeled with I-125 using chloramine-T. The labeled ribosomes, after treatment with glutaraldehyde, were injected into rabbits either subcutaneously (through the dorsum of the foot) or intravenously (through the ear). After subcutaneous injection, 40% of the activity accumulated in the lymph nodes during the first 5 hr, and the 70S ribosomal particles were shown to remain within the lymphatic system for at least 8 hr. After intravenous injection, 71% of the activity was detected in the liver within minutes by scintigraphic techniques. The effective half-time of the label in the liver from glutaraldehyde-treated I-125-tagged 70S ribosomal colloidal particles is 4-5 hr. No pyrogenic response was observed. Barring any deleterious side effects, the results indicate that biologic cell components of definite dimensions (in this case E. coli 70S ribosomes about 20 nm in diam) could be considered as radiocolloids for lymph-node and liver imaging.


Trace contamination of anesthetic gases: a brief review. Data have been reviewed relevant to the effects of chronic exposure to trace quantities of inhalation anesthetic agents. Much circumstantial evidence exists indicating a probable hazard to the clinician administering these agents. However, all previous studies in man have not identified the potential hazard of nitrous oxide without the presence of other contaminating agents.


Restoration of fractured and hypoplastic incisors by the acid etch resin technique: a three-year report. A total of 724 fractured and hypoplastic incisors were restored using an acid etch resin technique in combination with a newly developed "chamfer shoulder" preparation. The retention rate of the restorations, as observed over a three-year period, was 97.1% for the incisal restorations and 94.4% for the labial veneer restorations. According to evidence reported here, the use of pins for such cases is not recommended.


Free neurovascular flap from the first web of the foot in hand reconstruction. To identify an anatomically reliable and functionally acceptable neurovascular free flap for use in hand reconstruction, 50 fresh cadaver feet were dissected under the operating microscope, with particular attention paid to the anatomy of the first web area. A distal communicating artery was seen in 100% of dissections, allowing either dorsal or plantar donor artery inflow to nourish the entire flap area. Because of the ease of dissection, the first dorsal metatarsal or dorsalis pedis is suggested as the donor artery, and a dorsal branch of the greater saphenous venous system is suggested as the donor vein. The deep peroneal nerve was seen to consistently innervate the first web and, along with the plantar digital nerves, is suggested as an anatomically identifiable donor nerve. Either part of the foot first web may be used alone or together as a free flap. When indicated further dorsal skin may be incorporated into the web flap to expand its application. Two-point discrimination studies of the lateral plantar surface of the great toe in 50 normal individuals showed an average of 11.2 mm. This was significantly better as a potential donor flap than the medial dorsum of the foot where the average was 32 mm. A single case demonstrating the application of this flap in hand reconstruction is presented.


Volume increase by Z-plasty to the finger skin: its application  in electrical ring burns. Arc welder finger ring burns are an illustration of traumatic annular contractures. The potential for these and similar circumferential injuries to produce digital edema has led us to utilize Z-plasty in the early wound healing phase. Early Z-plasty in selected cases can prevent this loss of function from edema.


Brentano's Psychology from an Empirical Standpoint and Freud's early metapsychology. Brentano's Psychology from an Empirical Standpoint and Freud's early metapsychology, particularly the Project for a Scientific Psychology, show some interesting similarities. In psychological theory, both men stressed the motivated nature of thought and a conception of a "psychological reality" that is superior to "material reality." Both saw consciousness of one's own mental activity as arising retrospectively and having a calming effect on emotions. Both emphasized a process of "judgment" or "reality testing," made possible only by the presence of a strong unity of consciousness of ego. Methodologically, both agreed that the retrospective analysis of subjective experience is the principal tool of psychology. Freud eventually agreed with Brentano that psychology proceeds best when separated from physiology. They applied similar criteria to "proofs" of the unconscious, with Brentano concluding negatively, and Freud, positvely, after considering new evidence. This article considers the similarities and concludes that they suggest some sort of influence of Brentano on Freud.


Franz Boas and his plans for an International School of American Archaeology and Ethnology in Mexico. The expansionist policy of the United States at the turn of the century widened the horizons of American anthropology. The International School of American Archaeology and Ethnology was one of the first attempts by American anthropologists to carry out systematic research in foreign lands. Motivated partly by a wish to strengthen the quality of American anthropology, Franz Boas succeeded in gaining the cooperation of several European and American institutions. The purpose of the school was to conduct rigorous anthropological investigations in Mexico. Obsessed with professionalizing the discipline, Boas failed to take into account the turbulent political climate of Mexico when planning the school. Although it did good work for a number of years (1910-1914), the school was broken up forever in 1914 because of the Mexican revolution. Attempts at resurrection failed for numerous reasons.


In defense of a critical-presentist historiography: the fact-theory relationship and Marx's epistemology. It is argued that mainstream social scientists made an "error" during the crucial formative years of the 1920s and 1930s. This historical error involved relinquishing, or at the very least ignoring, some of the epistemological views within the social sciences concerning the fact-theory relationship as developed by Marx and others in his tradition. The contemporary nature of some of Mark's epistemic views is demonstrated, where the case is made that some recent developments in the philosophy of science are converging with some of his original insights. The methodological approach of the article--judging the past in the context of the present--is defended on the basis of Marxian historiography, especially as developed by those noe-Marxists of the Frankfurt School. In the latter discussion "justificationist presentism" is criticized from the perspective of a "critical presentism."


Joseph Lancaster and behavior modification in education. In the early 1800s, Joseph Lancaster (1778-1838) developed an inexpensive method of education large numbers of the poor in England. Referred to as the "monitorial system," his method utilized students as monitors who performed many of the tasks normally undertaken by a teacher; it was used to teach classes that often exceeded a thousand students at one time. Student monitors were responsible for teaching and evaluating small groups of individuals in a somewhat regimented fashion. An incentive system was also used in a manner that closely resembles current applications of positive reinforcement in educational settings. Lancaster devised a token economy in which students earned tangible reinforcers for academic performance and deportment. Interestingly, the system spread quickly in the 1800s, and was implemented in many countries throughout the world. Its use declined as a result of both specific sources of dissatisfaction, such as the lack of individualization and dependence upon rote memorization, and general changes in the philosophy of education.


Immunocytochemical localization of epidermal growth factor in mouse submandibular gland. The cellular and subcellular localization of epidermal growth factor in the submandibular glands of male and female adult mice was established by immunoperoxidase techniques. In light microscopic preparations epidermal growth factor was found exclusively in the granular convoluted tubules of the gland. The intensity of staining for epidermal growth factor varied from cell to cell, and some cells apparently were negative. The pattern of staining was similar in the glands of male and female mice; however, the granular convoluted tubules are androgen-responsive, and thus more extensive and composed of larger cells in males. In thin sections epidermal growth factor was most heavily concentrated in the secretion granules of the granular convoluted tubule cells. Within a given cell there was variation in intensity of staining of individual secretion granules, with some granules appearing minimally reactive or negative. The only other cell component with deposits of reaction product was the ribosomes.


The unlabeled antibody enzyme method. Attempted use of peroxidase-conjugated antigen as the third layer in the technique. The use of covalently bound peroxidase-immunoglobulin conjugate has been compared with soluble peroxidase-antiperoxidase complex as antigen in the third layer of the unlabeled antibody enzyme method. The conjugate conferred no staining. The low sensitivity of the conjugate appeared to be due in part to interference by unconjugated immunoglobulin and in part to diminution of antigenic reactivity of immunoglobulin as a result of conjugation. The triple specificity amplification inherent in the unlabeled antibody enzyme method with peroxidase-antiperoxidase complex may be lost if a peroxidase-immunoglobulin conjugate could be prepared in such a manner that staining occurs.


Immune complex receptors on cell surfaces. III. Topography of macrophage receptors demonstrated by new scanning electron microscopic peroxidase marker. Receptors for immune complexes have been localized on rabbit alveolar macrophages with scanning electron microscopy by exposing the cells first to a soluble immune complex composed of horseradish peroxidase and antibody to horseradish peroxidase, and then incubating with a benzidine-containing substrate that yields crystalline reaction product. Receptors were visualized by this means as sites of attachment of laminated slender crystals that were easily distinguished from macrophage surface structures. Receptors appeared most abundant on cytoplasmic veils and pseudopods and in the perinuclear region of macrophages minimally spread over the coverslip. Further macrophage spreading was associated with lighter receptor staining.


Evaluation of the utility of air-dried whole mounts for quantitative electron microprobe studies of platelet dense bodies. A variety of electronmicroscope techniques have been used to examine how the air-drying process may affect the dense bodies in whole mounts of platelets. (a) Selected-area-diffraction and electron microprobe studies suggest that the air-drying process can result in the formation of crystalline precipitates of sodium chloride on grid films and platelets. However, no crystals were detected in the calcium-and-phosphorus-containing matrix of dense bodies. (b) Tilting studies show that dense bodies in human platelets are spherical or ellipsoidal in shape. Dense bodies in rabbit platelets, in contrast, appear flattened in a horizontal plane. (c) Human-platelet dense bodies probed with a small (20 nm diameter) spot vary widely in their peak/background ratios for calcium and phosphorus-a finding that suggests that the two elements may not be evenly distributed throughout the dense-body matrix. Nevertheless, when dense bodies are probed with a larger (200 nm diameter) spot, they do not appear to differ appreciably among themselves in their calcium or phosphorus content. The data suggest that with human platelets, air drying may be a preparative procedure which permits comparison by microprobe techniques of dense-body matrix content in platelet populations.


A review of research examining the coronary-prone behavior pattern. With the increasing incidence of coronary heart disease (CHD) and the subsequent interest in factors which contribute to its development, the idea of a coronary-prone (Type A) behavior pattern has attracted much attention in recent years. In this paper, research on the coronary-prone behavior pattern is reviewed. Type A behavior is found to be fairly accurate in predicting the likelihood and severity of CHD, at least in large groups of people. Problems in measuring and in more clearly defining the components of Type A behavior are discussed. Future investigations are urged, focusing on how the behavior pattern develops and on strategies for intervention.


In vivo transfer of an R factor within the lower gastro-intestinal tract of sheep. The transfer of an R factor from donor E. coli introduced into the rumen of adult sheep to strains of the coliform microflora resident post rumen in the lower gastro-intestinal tract was found to be greatly increased when the animals were subjected to a short period of starvation (ca. 24-48 h). This also resulted in coliform organisms containing the resistance determinants of the R factor being excreted for much longer periods, sometimes for months afterwards. As no antibiotic treatment was given to the animals during these experiments, possession of the R factor should have conferred no selective advantages on the host cells and other plasmids could possibly be transferred similarly in vivo in sheep or other ruminants and perhaps also within the gut of monogastric animals.


Complement receptors in human renal glomeruli. Further evidence by immunofluorescence. A new method is described for demonstrating the presence of glomerular receptors for the third component of complement in human kidney. Frozen sections are incubated together with normal human serum and inulin. Activated C3 is detected by a fluoresceinated anti-C3 antiserum. Glomeruli are labelled by C3-coated inulin particles while there is no labelling in tubules and interstitium. This method is easy and rapid and it allows the exact localization of C3 glomerular receptors.


An improved plaque assay for mouse myeloma (MOPC 315) cells for use in studies of humoral and cell-mediated immunity. Dinitrophenyl-bovine albumin was coupled at room temperature to sheep red blood cells in a procedure which minimized spontaneous lysis and allowed the preparation of large batches and their use for at least 3 weeks. The modified erythrocytes were used as a substrate for detecting local hemolytic plaques in agar by myeloma MOPC 315 cells, which secrete a paraprotein IgA with high affinity for dinitrophenyl ligand. Conditions maximizing the number of plaques formed by a given number of tumor cells were found to include coupling the erythrocytes at 1 mg/ml dinitrophenyl-bovine albumin with a molar ratio of about 50, and incubation with an amino-to-carboxy cross-linking agent, 1-ethyl-3(3 dimethyl aminopropyl) carbodiimide, at 2 mg/ml for 50 min. The method thus developed was employed to measure cellular and antibody-dependent immune reactions against the MOPC 315 cells. The experimental results show comparisons of the plaque technique with other measurements of tumor cell injury. The nature of the assay, which requires only 500 cells per plating, and which tests the synthetic capacity of single cells, suggests its use in experiments which limit the number of target cells, and in immune reactions causing injury, but not necessarily lysis, of the target cells.


Quantitative in vitro phagocytic rate measurements. A model for the calculation of phagocytic rate constants based on the assumption that the process of phagocytosis can be treated in a manner analogous to chemical reactions of the first order is suggested. By employing such an approach, accurate rate constants of ingestion of in vitro cultivated macrophages can be obtained following minimal time intervals from the onset of phagocytosis.


An immunofluorescent technique for the detection of lymphocyte alloantigens. A triple-layer immunofluorescent technique for the detection of cell surface alloantigens has been developed. Chicken lymphoid cells were treated in sequence with: 1) chicken alloantibody, 2) rabbit anti-chicken Fc or anti-gamma chain serum and 3) FITC-labelled sheep anti-rabbit globulin. The second layer was diluted to that point where normal serum treated lymphocytes were no longer stained but without diminishing the degree of specific staining. This method enables the study of alloantigens expressed on B cells, avoiding the problem of staining of surface Ig-bearing lymphocytes.


The mouse circus: a simple apparatus for thoracic duct cannulation and continuous intravenous infusion. A simple apparatus is described for maintaining mice during thoracic duct cannulation and/or continuous intravenous infusion, which involves much less restraint than current methods. An Appendix describes the techniques of intravenous cannulation which have been used for prolonged infusion or cardiac catheterisation.


Immunochemical determination of ganglioside GM2, by inhibition of complement-dependent liposome lysis. Immunochemical quantitative determination of a lipid antigen, ganglioside GM2, has been developed, based on the inhibition of the immune lysis of liposomes containing the antigen in their lipid bilayer. It has been shown that the full expression of the antigenicity of the competing lipid requires its dispersion in accessory lipids. The assay of inhibition of liposome lysis can be used also for the establishment of the antigenic similarity of structurally related lipid antigens.


Demonstration by labeled treponemal antigen of specific antibodies in the tissue infiltrates of secondary syphilis. Specific antitreponemal antibodies have been demonstrated by immunofluorescence techniques in the lymphoplasmocytic infiltrates which characterize early symphilitic lesions. A purified suspension of Nichols strain Treponema pallidum was sonified and labeled with fluorescein isothiocyanate and applied to cryostat sections of 12 biopsy specimens from the cutaneous lesions of 11 patients with proven secondary syphilis, using a modified direct immunofluorescence procedure. Specimens from various inflammatory dermatoses processed similarly served as controls. Granular fluorescence was noted in the dermis in 9 of the 12 specimens corresponding to areas of heavy plasma cell infiltration and some fluorescence was found directly on plasma cells which were identified by subsequent hematoxylin and eosin staining. This fluorescence could be blocked by prior incubation of the sections with unlabeled sonified treponemal suspension. Control slides did not reveal any fluorescence. The use of labeled treponemal antigen may aid the tissue diagnosis of early syphilitic lesions which can mimic a variety of dermatological disorders.


The effect of the divalent cation ionophore A23187 on amphibian melanophores and iridophores. Calcium ions are required in the medium for the full darkening action of melanocyte-stimulating hormone, but not for its second messenger, cyclic adenosine monophosphate, on isolated frog skin. Thus, the possible effect of the divalent cation ionophore A23187 was studied, using the skin of Rana pipiens. It is a potent darkening agent over the range of 1-2 micrometer, as measured by reflectance change and microscopic observation. During the darkening response of A23187, melanosome dispersion takes place in both dermal and epidermal melanophores, as well as aggregation of iridophore reflecting platelets. The darkening is slowly reversible in Ringer's solution, thus it is not due to toxicity. The darkening is partially dependent on the presence of calcium in the medium under certain conditions, suggesting that the response involves both the uptake and release of calcium ions. Cytochalasin B reversibly inhibits the darkening produced by A23187. Since this drug is known to bring about the breakdown of microfilaments, its inhibitory action is in accord with the concept that the ionophore is stimulating microfilament function. A23187 administered to tissue cultured embryonic salamander (Ambystoma maculatum) melanophores produced an irreversible rounding up of the cells, suggesting a toxic effect. The results with frog skin are interpreted as supporting the concept that the action of melanocyte-stimulating hormone involves the production of pigment granule movements as a result of the interaction of calcium ions with intracellular microfilaments and possible also the breakdown of microtubules. The mobilization of calcium ions could be brought about by cyclic adenosine monophosphate.


Circulating IgA anti-basement membrane antibodies in linear dermatitis herpetiformis (Duhring): immunofluorescence and immunoelectronmicroscopic studies. IgA deposits were observed by direct immunofluorescence in linear distribution along the basement membrane zone in a case of dermatitis herpetiformis (Duhring). In addition, in the serum of the same patient circulating IgA antibasement membrane zone antibodies were detected by indirect immunofluorescence, utilizing normal human skin and monkey esophagus as substrates. The ultrastructural localization of in vivo-bound IgA and circulating IgA antibasement membrane zone antibodies fixed to substrate tissue in vitro was found to be in the uppermost strata of the dermis below the basal lamina.


Comparison of Escherichia coli from bacteriuric patients with those from feces of healthy schoolchildren. The properties of 709 strains of Escherichia coli isolated from feces of healthy schoolchildren were compared with those of 115 strains from the urine of girls with asymptomatic bacteriuria (ABU) detected in a screening program. These fecal strains were also compared with 45 strains that caused asymptomatic reinfections and 10 that caused symptomatic reinfections in the same group of girls. Typing of O antigen was done by direct bacterial agglutination, and K typing was done with a serum agar technique. Hemolytic capacity was assessed in solid medium. Sensitivity to the bactericidal effect of normal serum and minimal inhibitory concentrations of ampicillin were also determined. The strains isolated from girls who had reinfections of ABU were found to be a random sample of the fecal flora, but the strains from children with symptomatic reinfection were not. Strains from index patients with ABU differed from the other groups in a way that was indicative of adaptive changes in the structure of cell envelopes.


Antibodies to Candida albicans in IgA-deficient humans. Titers of serum antibody to Candida albicans were studied in 111 IgA-deficient subjects by the passive hemagglutination method. Circulating antibodies to C. albicans (mean titer, 1:7) were found in only 7.3% of the 111 subjects, as compared with 80% of 137 controls (mean titer, 1:16). However, two IgA-deficient patients with systemic candidiasis were positive for these antibodies (mean titer, 1:64). Indirect immunofluorescence tests indicated that the antibody response in IgA-deficient subjects to superficial or deep-seated infection with C. albicans consists mainly of immunoglobulins of the IgG rather than of the IgM class.


Studies with a new generation of oral attenuated shigella vaccine: Escherichia coli bearing surface antigens of Shigella flexneri. In an attempt to develop a safe, proliferating, oral, attenuated vaccine against shigellosis, genes that control the synthesis of group- and type-specific somatic antigens of Shigella flexneri 2a were transferred via conjugation to a recipient strain of Escherichia coli. The resultant hybrid (E. coli expressing shigella surface antigens) vaccine strain, PGAI 42-1-15, believed to have a complete (smooth) lipopolysaccharide, was given to volunteers in two vaccination-challenge studies. The vaccine was well tolerated and gave evidence of intestinal proliferation. In trial no. 1, volunteers given two doses of vaccine one month apart were challenged after eight weeks with 10(4) virulent S. flexneri 2a. Attack rates were comparable in vaccinees (50%) and controls (40%). In trial no. 2, vaccinees were given three weekly doses of vaccine and were challenged four weeks later with a small inoculum (10(2)) of S. flexneri 2a. Again, attack rates among vaccinees (47%) and controls (39%) were similar. It is unclear why this theoretically ideal, live shigella vaccine failed to protect against S. flexneri 2a.


Transfer of multiple antibiotic resistance from Bacteroides fragilis to Escherichia coli. Multiple antibiotic resistance was transferred from a clinical isolate of Bacteroides fragilis to a strain of Escherichia coli K12. Resistance to ampicillin, amoxicillin, cephalothin, tetracycline, minocycline, and chloramphenicol was transferred as a unit, but the resistance markers became segregated during storage of the recipient strains.


Evaluation of some of the parameters of the enzyme-linked immunospecific assay. In an attempt to standardize the procedure for the enzyme-linked immunospecific assay, several parameters were examined. It was determined that horseradish peroxidase was the enzyme of choice and that several brands of polystyrene tubes and plates could be used. The test was considerably shortened by omitting long incubation steps previously used for eliminating background fixation of conjugate. The reproducibility of the procedure proved to be excellent, but reactivity curves indicated that use of a single dilution is not adequate for quantitative tests. A "standard" procedure was proposed.


Host heme catabolism in drug-sensitive and drug-resistant malaria. Chloroquine resistance has arisen in both human and murine forms of malaria. CR Plasmodium berghei in mice does not produce the malaria pigment which is characteristic of the CS form. Determinations of carbon monoxide production (i.e., host heme catabolism) by individual mice revealed that those infected with CS P. berghei produce only one fourth as much carbon monoxide as do CR infected mice at all levels of infection. These observations confirm the idea that malaria pigment is composed of precipitated host cell hemoglobin and suggest that drug resistance is accompanied by a basic alteration in parasite-mediated hemoglobin catabolism.


Role of vitamin E in glutathione-induced oxidant stress: methemoglobin, lipid peroxidation, and hemolysis. Red blood cells (RBC) from normal and vitamin E-deficient rats were incubated in a hypertonic solution of reduced glutathione adjusted to pH 8. Methemoglobin formation occurred in intact RBC from both normal and vitamin E-deficient rats. Hemolysis was significantly greater in RBC from vitamin E-deficient rats. Experiments with catalase, superoxide dismutase, and methional showed that H(2)O(2) was the primary extracellular source of oxidant stress. Extracellular superoxide and hydroxyl radical were not involved in oxidant stress. Experiments with dimethyl sulfoxide showed that intracellular hydroxyl radical, generated from H(2)O(2), was the hemolytic agent. Neither methemoglobin formation nor lipid peroxidation involved hydroxyl radical. Indeed, lipid peroxidation and hemolysis in RBC from vitamin E-deficient rats were concurrent rather than consecutive events. Phase contrast microscopy showed that rigid, crenated RBC with a precipitate around the interior periphery formed during glutathione-induced oxidant stress. The precipitate dissolved slowly as the crenated RBC were converted to smooth ghosts. It appeared that protein precipitates involving mixed disulfide bonds were reduced and solubilized when extracellular glutathione penetrated the ruptured cell. Comparisons between normal RBC and vitamin E-deficient RBC suggest that vitamin E has little effect on the inward diffusion of extra-cellular H(2)O(2). Vitamin E apparently interacts with different oxidant species derived from intracellular H(2)O(2) in preventing lipid peroxidation and the sulfhydryl group oxidation leading to hemolysis.


Immunocytochemical studies for the localisation of measles antigens in multiple sclerosis plaques and measles virus-infected CNS tissue. Actively demyelinating central nervous system (CNS) lesions from a patient with acute multiple sclerosis (MS) were tested for measles antigens using peroxidase-conjugated antimeasles antibody. No evidence of measles antigens was found. Similarly reacted tissue from 2 patients with chronic MS also revealed no evidence of measles antigens. Identically treated and simultaneously tested measles-infected CNS cultures and human SSPE brain tissue stained strongly for measles antigens. The possible reasons underlying the failure to detect measles antigens in MS are discussed.


Immunofluorescence study of lymphocytic infiltration in gliomas. Identification of T-lymphocytes. Five human brain tumours (3 glioblastomas and 2 astrocytomas) and 5 rat brain tumours induced in Sprague--Dawley animals by systemic administration of N-methyl-N-nitrosourea (3 pleomorphic gliomas and 2 mixed gliomas) were studied. The human brain tumours were surgical specimens excised from patients with no cranial surgery prior to their disease. The experimental brain tumour had been adapted to tissue culture, propagated in vitro and then transplanted to immunocompetent and immunodeficient rats of the same stock. The above-described material was selected in consideration of the mononuclear cell infiltrates occurring in these tumours. Frozen sections of human and rat gliomas, the latter both primary and transplanted, were prepared and investigated as to the presence of T-lymphocytes within the mononuclear round cell infiltrates. This was done with the indirect immunofluorescence method using rabbit antisera against man and rat T-lymphocytes. With this technique a variable percentage of T-lymphocytes was demonstrated in the cell infiltrates of human and rat gliomas alike. The tumour transplanted in thymectomized rats showed only isolated, scattered, positive-reacting cells, i.e., cells recognizable as T-lymphocytes by the above method. The results can be interpreted as circumstantial evidence for the occurrence of tumour-specific and/or tumour-associated antigens in the parenchymal cells of spontaneous and chemically-induced gliomas.


Immunochemical comparison of synaptic plasma membrane and synaptic vesicle membrane antigens. A synaptic vesicle fraction and a synaptic plasma membrane fraction obtained after subfractionation of synaptosomes from chick forebrain have been used to produce antisera in rabbits. Immunofluorescence histology with the two antisera revealed that they reacted strongly with synaptic terminal regions present in the chick forebrain, cerebellum and spinal cord. In addition, the synaptic plasma membrane antiserum (but not the synaptic vesicle antiserum) reacted with preterminal axons in the cerebellum and spinal cord. Comparison of the two antisera by two-dimensional immunoelectrophoresis, revealed the presence of common antigens in the synaptosomal vesicle and plasma membrane fractions. Incubation of synaptosomes in vitro with the synaptosomal vesicle antiserum and complement produced a dose-dependent inhibition of synaptosome swelling up to a maximum of 55% of that obtained with the synaptosomal plasma membrane antiserum. The results of this test are consistent with the hypothesis that some synaptosomal vesicle antigens may be present also in the synaptosomal plasma membrane and imply that they face the external surface of the synaptosomes. The fate of vesicle membrane components in synaptosomal plasma membranes is not known. The possibility is discussed that they may be recycled locally by a mechanism similar to that proposed by Heuser and Reese (1973) for re-use of synaptic vesicle membranes at the neuromuscular junction.


Mouse spinal cord in cell culture. I. Morphology and intrinsic neuronal electrophysiologic properties. 1. Reliable methods for establishing fetal mouse spinal cord (SC) and dorsal root ganglion (DRG) cells in long term (greater than 1 mo) dissociated cell cultures are described. These cells have been studied by morphologic and intracellular electrophysiologic techniques. 2. Cells studied electrophysiologically can be relocated after preparation for electron microscopy and examined in thin sections. The electron microscope shows that the surface membranes of these cells were directly accessible to the culture medium. The surfaces of SC cells were studded with synaptic boutons, whereas the DRG cell surfaces generally had none. 3. Current-voltage relationships and linear electrotonic properties of the neurons are described. Delayed and anomalous rectification were seen in both cell types. The length of SC cell dendrites was about one characteristic electrotonic length, while little or no contribution of the relatively sparse DRG cell processes was seen in the transient responses of the DRG cells. 4. Postspike and posttetanic hyperpolarizations in DRG cells were due to a surface membrane conductance increase; this was probably primarily an increase in K+ conductance. Post-activation hyperpolarization in SC cells was primarily due to activation of an electrogenic Na+ pump.


Bilateral persistent trigeminal arteries. Case report. A case is presented of bilateral persistent trigeminal arteries associated with reticulum cell sarcoma of the cerebellum.


Comparison of fascicular, interfascicular and epineural suture techniques in the repair of simple nerve lacerations. Simple lacerations of dog anterior tibial nerves were repaired utilizing fascicular interfascicular, and epineural suture techniques. Two months following repair, the involved regions of the nerve were removed and stained with hematoxylin and eosin, Weil, Bielschowsky, and Masson trichrome stains. An independent, unbiased observer rated proximal and distal myelin, connective tissue responses, and proximal to distal axon counts. The proportion of proximal to distal axons revealed no significant difference between fascicular (77%) and epineural (67%) neurorrhaphy, but showed a significant beneficial effect of fascicular suture to the interfascicular (52%) repair (significance at less than 0.02). Masson stain revealed dense connective tissue responses, but the axon counts were adversely affected only when the separate fascicles showed internal disruption by connective tissue. In general, distal myelinization was mildly superior with the fascicular neurorrhaphy technique. It appears that in simple lacerations in nerves which are repaired via direct end-to-end suture, there is no significant advantages to fascicular over epineural repair; however, there is a definite deleterious effect of interfascicular neurorrhaphy. The theoretical and technical implications of these findings are discussed.


Management of upper midfacial injuries.

Management of upper midfacial injuries. Midfacial fractures may extend into orbital roofs, frontal sinus walls, and ethmoidal and sphenoidal bones if the traumatic impact is of sufficient force. The cranium and facial skull are affected and there is the risk of intracranial complications. Management thus requires a teamwork approach. Isolated compound fractures of the anterior frontal sinus may be reconstructed by direct access through the wound. We prefer a coronal hairline incision in most cases. In isolated (compound) fractures of the orbital roof, combined frontomaxillary fractures without dural involvement, and frontobasal fractures that necessitate an intracranial approach for dural closure, primary upper midfacial reconstruction is attempted. In frontal comminution, preservation of the frontal bone or orbital roof is impossible. Secondary management is indicated after an interval of at least six months for fronto-orbital reconstruction with autologous or alloplastic material.


Application of physiologic and prosthodontic principles in reduction of mandibular fractures. A review of basic prosthodontic principles and the muscle physiology on which they are based is presented. Proper reduction of fractures of the edentulous mandible and maxilla requires the incorporation of a correctly determined freeway space into either the patient's preexisting dentures or into the split Gunning-type splints.


Skin reactivity to Candida and streptokinase-streptodornase antigens in normal pediatric subjects: influence of age and acute illness. Tests for delayed cutaneous hypersensitivity were carried out with Candida and streptokinase-streptodornase antigens in 245 normal pediatric subjects, ranging in age from 17 days to 5 10/12 years. The group comprised patients with a variety of acute illnesses and a series of control subjects. There was a progressive increase in the frequency of cutaneous responses with age. About one-third of infants under one year and four-fifths of one to five-year-old children had positive induration reactions to at least one antigen. Candida was the more reactive antigen in the first year of life; thereafter there was comparable responsiveness to both antigens. Our results showed no difference in percent reactivity between acutely ill and control subjects.


The differential leukocyte count in the assessment and outcome of early-onset neonatal group B streptococcal disease. The usefulness of the differential white blood cell count in distinguishing early-onset group B streptococcal disease from other causes of neonatal respiratory distress was studied in 45 infants with culture-proved infection. The initial diagnosis was hyaline membrane disease in 19 infants, wet lung syndrome 13, and other causes of respiratory distress in 13. Thirty-nine (87%) had abnormal absolute neutrophil counts, 25 with neutropenia and 14 with neutrophilia. The absolute immature neutrophil count was elevated in 19 infants (42%). Forty-one infants (91%) had an abnormal immature neutrophil to total neutrophil ratio. All infected infants were identified when both the absolute total neutrophil count and ratio were used. The differential white cell count appears to be a useful tool for screening infants presenting with respiratory distress in the first 48 hours of life and for separating early-onset group B streptococcal disease from other causes of neonatal respiratory distress.


Detection and quantitation of bacteremia in childhood. Quantitative blood cultures were sought in 383 children, from whom routine blood cultures were obtained because of fever, by direct plating of 10 and 100 microliter blood onto solidified media. There were 14 positive cultures from 12 patients. These were 7 Hemophilus influenzae type b, 5 Streptococcus penumoniae, and 2 Staphylococcus aureus. The direct-plating technique permitted more rapid identification of positive cultures, and detected three episodes not identified by routine broth culture. Bacterial counts ranged from 20 to greater than 10(4) bacteria/ml blood. In the three cases of H. influenzae type b meningitis, bacteremia exceeded 10(3)/ml. Among nine patients in whom bacteremia was unassociated with meningitis, (bacteremia without evident localized disease 5, pneumonia 2, epiglottitis 1, peritonitis 1), bacteremia was less than 10(3)/ml. This technique may aid detection of bacteremia and help identify those children at highest risk for developing septic complications, such as meningitis.


Pathologic and immune factors in thyroid disease. Thyroid glands from 33 children with hyperthyroidism and nine with juvenile lymphocytic thyroiditis were examined histologically and for IgG, IgA, IgM, and C3 by immunofluorescent staining. There was no significant difference between glands with JLT and those with hyperthyroidism in the degree of lymphoid infiltration or lymphoid follicle formation. In thyroiditis there was no correlation between the degree of histologic abnormalities and the presence of immunofluorescent staining for IgG, IgM, or IgA. In hyperthyroidism there was a correlation between the degree of histologic abnormalities and the presence of IgG. In both groups of patients LI and LFF were distinctly more severe in glands positive for C3. Postsurgical hypothyroidism correlated with LI but not with LFF, IgG, or C3.


Controlled trial of bicarbonate therapy in high-risk premature newborn infants. Sixty-two high-risk acidemic premature newborn infants, given maintenance intravascular infusions of 10% glucose, were assigned to liberal or restricted sodium bicarbonate treatment groups. Those infants in the liberal group received 5 to 15 mEq bicarbonate/dl 10% glucose, depending on the degree of acidosis. Among infants given bicarbonate, correction of pH was not more rapid and mortality was not decreased. Instead, there was a small increase in the number of deaths, but the incidence of intraventricular hemorrhage was similar to that in infants not given bicarbonate.


The effect of independent variations in inspiratory-expiratory ratio and end expiratory pressure during mechanical ventilation in hyaline membrane disease: the significance of mean airway pressure. The specific aspects of the respiratory cycle during mechanical ventilation that allow for optimum gas exchange are still controversial. To further clarify the relationship of inspiratory:expiratory ratio and positive end expiratory pressure to optimum ventilation and oxygenation, five premature lambs with severe hyaline membrane disease were ventilated with volume-present infant ventilators at I:E ratios of 1:4 and 1:1 and PEEP levels of 0, 5, and 10 cm H2O. For each I:E ratio/PEEP combination, pH, Pao2, PaCO2, PAO2, PACO2, peak  inspiratory pressure, mean inspiratory pressure, and mean airway pressure were measured and compared. Optimum ventilation and oxygenation were related to MAP, but not to I:E ratio, PIP, or MIP. As MAP increased from 6 to 14 cm H2O, progressive improvement in Pao2, PaCO2 (A-a) DO2 and (a-A) DCO2 was evident. Above 14 cm H2O, there was progressive deterioration in these measurements. There was also a direct relationship between MAP and mean pleural pressure. These results indicate that during mechanical ventilation there is an optimum MAP at which gas exchange is best. Since MAP changes with any change in PIP, PEEP, or I:E ratio, it provides a useful composite measure of all pressures transmitted to the airways by the ventilator.


Comparison of tolmetin sodium and aspirin in the treatment of juvenile rheumatoid arthritis. The Pediatric Rheumatology Collaborative Study Group was established in 1973 to undertake systematic trials of new drugs in the treatment of juvenile rheumatoid arthritis. The first drug evaluated was tolmetin (1-methyl-5-p-toluoylpyrrole-2 acetic acid), a new nonsteroid anti-inflammatory agent. A four-week open trial with 30 patients and a subsequent 12-week double-blind trial against aspirin with 107 patients were conducted. Tolmetin and aspirin had equal anti-inflammatory and analgesic effects in the treatment of JRA. Elevations of transaminase values attributed to aspirin were not found with tolmetin. Adverse effects accompanying administration of tolmetin did not appear to be of major clinical significance.


Effect of topical application of phenylephrine hydrochloride on hyperplastic gingivitis. A double blind study was conducted to evaluate the effectiveness of Phenodent Type A (brand of phenylephrine hydrochloride) on decongesting hyperplastic gingivitis. Three solutions were used: a 0.5% a placebo, and a 0.25% concentration of phenylephrine hydrochloride. The periodontal disease index was used to score variables which might have an effect on gingival response to local irritants. Impressions were taken and casts were made on 45 subjects at 0, 1, 3, and 6-week intervals. An instrument with accuracy of 0.001 inch was constructed to measure the changes in the interdental papillae of the stone casts. No significant reduction of gingival volume was established for any of the three solutions.