Function of S-adenosylmethionine in germinating yeast ascospores. Germination and outgrowth of ascospores of Saccharomyces cerevisiae 4579 require both methionine and adenine, whereas leucine is only required for outgrowth. The methionine requirement may be satisfied by S-adenosylmethionine, but this sulfonium compound will not substitute for adenine. Between 30 and 70 min of protein synthesis is initially required for the completion of germination in strain 4579. The inhibition of S-adenosylmethionine synthetase by trifluoromethionine prevents both germination and protein synthesis. During the initial stages of germination, the S-adenosylmethionine synthetase, S-adenosylmethionine decarboxylase, and transfer ribonucleic acid methyltransferases increased significantly, indicating that polyamines and/or the methylation of transfer ribonucleic acid are required for the initiation of germination.


Expression of R-plasmid functions during anaerobic growth of an Escherichia coli K-12 host. The normal habitat of enteric bacteria is largely anaerobic. Expression of the three characteristic properties of R-plasmids, drug resistance, vegetative replication, and fertility, was therefore studied in Escherichia coli K-12 during anaerobiosis. Replication and drug resistance functions were not altered in the 45 R-plasmids studies, whereas the expression of fertility varied considerably among different R-plasmids during anaerobiosis. The R-plasmids could be divided into three groups, one showing a strong, the second a moderate, and the third little or no reduction of fertility by anaerobiosis. Plasmid-determined sensitivity to F-, N-, and I-specific phage, respectively, was well, although not absolutely, correlated with each of three groups mentioned. Anaerobiosis-aerobiosis appears to change the fertility of type F R-plasmids by influencing the degree of repression of their fertility functions such as the formation of sex pili. Although the minimum inhibitory concentrations of ampicillin, chloramphenicol, streptomycin, and tetracycline were unaltered by anaerobiosis, sulfonamide was found to be four- to eightfold less active under this condition in both resistant and sensitive strains. A surprisingly high frequency and uniformity of minimal inhibitory concentrations was observed for R-plasmid-mediated resistance to streptomycin and chloramphenicol.


Control of ribosome synthesis in Escherichia coli: analysis of an energy source shift-down. The rate of ribosome synthesis and accumulation in Escherichia coli during the transition after an energy source shift-down was analyzed. The shift was imposed on cultures of stringent and relaxed strains growing in glucose minimal medium by the addition of the glucose analogue alpha-methylglucoside. In the stringent strain, ribosome synthesis was almost instantaneously reduced after the shift, whereas the relaxed strain exhibited a more gradual response. The rate of messenger ribonucleic acid (mRNA) synthesis was affected similarly, though to a smaller extent. A comparison of the rates of synthesis and accumulation of ribosomal RNA (rRNA) and ribosomal proteins showed that far more ribosomal components were synthesized after the shift than were accumulated, indicating that a substantial part of the rRNA made after the shift was unstable. A new method was used to measure relative rates of rRNA synthesis and to estimate the transcription time for the rRNA operon under different conditions. In steady states of growth with growth rates ranging from 0.75 to 2.3 doublings/h, as well as during the transition after a shift-down, the transcription time of the rRNA operon was constant. The rate of synthesis of rRNA correlated during this transition - in contrast to the rate of accumulation (M. T. Hansen et al., J. Bacteriol. 122: 585-591, 1975) - with the ppGpp pool in the same way as has been observed during partial amino acid starvation.


R-plasmid transfer and its response to nalidixic acid. The conjugational transfer efficiency of 41 wild-type R-plasmids was studied in Escherichia coli K-12. Type I R-plasmids were transferred at comparatively high and rather uniform frequencies, whereas type F R-plasmids showed less uniform and, on average, somewhat lower transfer frequencies. R-plasmids not mediating sensitivity to F-, I-, or N-specific phages showed moderate transfer frequencies, and type N R-plasmids showed very low transfer frequencies. Various lines of evidence suggest that a well-expressed, but functionally inefficient, conjugation apparatus is the cause of the poor transfer of type N R-plasmids in liquid medium. Nalidixic acid efficiently inhibited transfer of type I and particularly type F R-plasmids, whereas the transfer of type N plasmids was resistant to the drug. Type F and type I plasmids appear to depend on at least one host function for their transfer, namely, the nalidixic acid-sensitive reaction in vegetative chromosome replication, whereas type N plasmids are independent of this function.


Release factor binding to ribosome requires an intact 16 S rRNA 3' terminus. Cloacin DF12 cleavage of Escherichia coli f[3H]MettRNA-AUG-ribosome complexes affects this substrate for in vitro peptide chain termination. Codon-directed release factors' (RF) 1 and 2 release of f[3H]methionine is inhibited by cloacin. Since cloacin inhibits RF1 and -2 binding to ribosomes but not RF-directed f[3H]methionine release from f[3H]met-tRNA-AUG-ribosome complexes when reactions contain 20% ethanol, we conclude that cloacin DF 13 inhibits formation of the termination codon recognition complex. Thus, cleavage of the 3'-OH 49-nucleotide sequence of the 16 S rRNA perturbs the codon-directed binding of RF to ribosomes.


Yeast hexokinase mutants.

Yeast hexokinase mutants. Using yeast mutants, it is shown that growth on glucose occurs even in the absence of both hexokinase A and hexokinase B; fructose growth requires at least one of these two enzymes. Expression of hexokinase A and of glucokinase seem to be regulated.


Studies of the lipid phase transitions of Escherichia coli by high sensitivity differential scanning calorimetry. High sensitivity adiabatic differential scanning calorimetry was performed on lipids, membrane vesicles, and whole cells of Escherichia coli enriched in particular unsaturated fatty acids by genetic means. Information concerning the shape of the transition is discussed. Transitions with an asymmetric shape reminiscient of a second order transition were observed. Comparison between the lipid transition observed in whole cells, membrane vesicles, and extracted lipids enriched in elaidate reveal some basic similarities. Studies of synthetic lipids were undertaken in an attempt to interpret the shapes of these transitions as a function of the lipid components of the membrane.


Novel properties of Escherichia coli exonuclease III. The specificity of hydrolysis of polynucleotide termini by Escherichia coli exonuclease III was studied with the use of oligothymidylate annealed to polydeoxyadenylate. The size of the products after 3' leads to 5'-hydrolysis of 5'-labeled substrate is temperature-dependent. At 25 degrees the enzyme can hydrolyze a polynucleotide chain up to the last 5'-terminal dinucleotide. A gradation of higher 5'-terminal oligonucleotides of defined chain lengths is produced after limit digestion by the enzyme when the temperature is raised between 25 degrees to 60 degrees. When the oligothymidylate was labeled at the 3'-ends with ribonucleotides, it was observed that exonuclease III can cleave a single or two consecutive ribonucleotides regardless of whether the ribonucleotides are base-paired or mismatched.


Ca2+ ions and the stimulation of 3-O-methylglucose transport by uncouplers in rat thymocytes. The uptake of 3-O-methylglucose by rat thymocytes follows a biphasic time course. 2,4-Dinitrophenol (10-3 M), carbonyl cyanide m-chlorophenylhydrazone (10-5 M) and oligomycin (5 microgram/ml) each reduce ATP levels in rat thymocytes by 85% and bring about 3- to 4-fold stimulation of the slow phase of 3-O-methylglucose uptake. No consistent effect is observed on either the half-time of the rapid phase of uptake or the relative proportions of the two phases of uptake in the presence of these agents. Ca2+ ions do not appear to play a necessary role in the stimulation of transport activity since cells depleted of exchangeable Ca2+ by treatment with the Ca2+-Mg2+ ionophore, A23187, plus [ethylenebis(oxyethylenenitrilo)]tetraacetic acid respond to uncouplers in exactly the same manner as untreated cells. The effect of dinitrophenol on the slow phase of 3-O-methylglucose uptake is reversible after 10 min of incubation. After 60 min however, cells washed free of dinitrophenol and incubated at 37 degrees exhibited an additional acceleration in transport activity. This stimulation of transport is characterized by an increase in the proportion of the rapid phase of uptake with little or no change in its half-time. The results suggest that rat thymocytes regulate their 3-O-methylglucose transport activity in two distinct fashions.


Study of the transfer RNAs coded by T2, T4, and T6 bacteriophages. T2, T4, and T6 bacteriophage tRNAs coding for arginine, leucine, proline, isoleucine, and glycine were isolated under conditions of short term and long term infection of Escherichia coli B cells. The corresponding phage tRNA species were examined for sequence homology by RNA-DNA hybridization analysis and by their relative behavior on reversed phase chromatography. The results indicate that all three T-even phages code for similar tRNA species; however, some tRNA species are homologous, others are not, and not all of the same tRNA species are coded by each bacteriophage. Reversed phase chromatography showed the presence of isoacceptor tRNAs for each phage aminoacyl-tRNA species. Pulse-chase experiments for [32P]tRNAGly suggest that the multiple isoacceptor species observed derive from the intracellular modification of a single tRNAGly gene product.


Amino acid sequence of the L-arabinose-binding protein from Escherichia coli B/r. The amino acid sequence of the L-arabinose-binding protein of Escherichia coli B/r was determined by sequenator analyses of reduced and S-pyridylethylated L-arabinose-binding protein and fragments derived by chemical and enzymatic cleavage of the native protein. The fragments were the products of cleavage by cyanogen bromide. BNPS-skatole, hydroxylamine, mild acid hydrolysis, limited trypsin digestion, chymotrypsin subdigestion, and subdigestion with Staphylococcus aureus protease V8. The COOH-terminal sequence was determined using bovine carboxypeptidases A and B and amino acid analyses. The L-arabinose-binding protein was determined to contain 306 amino acid residues, the sequence of which is presented below.


Evolution of the mandibular mesh implant. Between 1960 and 1972, the Dallas Veterans Administration Hospital Maxillofacial Research Laboratory developed and made over 150 cast-mesh implants. Successive designs were ovoid, circular, and double-lumened in cross section to improve implant strength, surface area for bioattachment, and adjustability. Sleeves, collars, and bows were employed in the assembly of these implants, with an acrylic condylar head attached when indicated. In 1972, our laboratory developed a mandibular mesh tray, cast in one piece on a single sprue, with preservation of the vertically adjustable ramus. Stainless steel replaced Vitallium because of its greater malleability. Essentially, a lost-wax technique is used to cast the mesh tray. The model of a mandibular segment is duplicated as  a refractory model. Mesh wax, made in our own custom-made die, is adapted to the refractory model. The unit is then sprued and invested. The wax is fired our of the mold in a gas furnace. Casting is done by the transferral of molten stainless steel from the crucible to the mold by centrifugal force in an electro-induction casting machine. Other mesh implants that have been developed are made from wire mesh, Dacron mesh, cast Ticonium, and hydroformed titanium.


Evaluation of wound-covering materials.

Evaluation of wound-covering materials. Physical and in vivo (burned rat model) evaluations as wound coverings were performed for 1) a freeze-dried collagen/poly (epsilon-caprolactone) (PCL) film laminate, 2) a freeze-dried PCL "foam"/PCL film laminate, and 3) a heat-dried collagen/PCL film laminate. Porcine skin and cadaver skin were also evaluated in vivo for the purpose of comparison. Water-vapor transmission rates and Young's moduli were measured. The degree of adherence of the coverings to the wound were measured. Grafts which became significantly adherent (greater than 150 dyne/cm2) to the wound within 1 day were most successful in promoting the formation of a viable tissue bed which appeared ready to accept further grafting. The force required to remove the PCL foam laminate from a full-thickness excision wound was found to increase from 170 dyne/cm2 on the first day postgraft to 1500 dyne/cm2 by the tenth day. The force required to remove freeze-dried collagen laminate remained constant at 200 dyne/cm2 over the 10 day test period. For the heat-dried collagen laminate, a force of only 50 dyne/cm2 was required on day 1, increasing to 200 dyne/cm2 on day 6. Insensible water-loss rates of animals grafted with the laminates were found to be similar to those from animals with human cadaver skin grafts and less than that from animals with porcine skin grafts. When moistened, the laminates prepared using the freeze-dried materials were flexible and somewhat transparent permitting observation of the wound.


Compression bone-plating: historical considerations. Compression of bone fragments to accelerate, or at least facilitate, bone healing was first applied in 1932 by Key to arthrodesis of the knee. Subsequently the same principle was applied to fractures, first by percutaneous pins as developed by Roger Anderson and then by implanted compression plates. Two types of compression plate have been evolved. One employs an axially oriented force produced by a separate screw or temporarily applied compression device, designed just for this purpose, as originated by Danis in 1949. The other, devised by me in 1956, is a so-called self-compressing plate, which generates compression force by inserting eccentrically placed chamfered screw-heads into slots in the plate so that the screws (as well as the bone fragments in which they are inserted) are translated toward each other. Numerous modifications of these two basic designs are now available.


Anterior cervical fusion. A review of thirty-three patients with cervical disc degeneration. Thirty-three patients who had undergone anterior cervical fusion for degenerative disc disease were reviewed to determine the efficacy of the procedure. Only patients who were available for examination and who had undergone operation at least one year previously were included in the review. Nearly all had had arm pain and three-quarters neck pain. Diminished neck movement and neurological abnormalities in the arms had been frequent findings. Diagnosis from the clinical features and plain radiographs is described. Myelography was not used routinely and discography was not used at all. Indications for operation and surgical technique are described. Results show that pain in the neck and arm was relieved in a high proportion of cases and that the neurological abnormalities often recovered. It is concluded that this operation is safe and has a definite place in the relief of pain from cervical disc degeneration resistant to conservative treatment.


Short term kinetics of luteinizing hormone secretion studied in dissociated pituitary cells attached to manipulable substrates. With the use of poly-L-lysine, a method has been developed which induces acutely dissociated rat anterior pituitary cells to attach to glass and polyacrylamide surfaces. In these attached cells the recovery of the secretory response, which is impaired in acutely dissociated cells, has been followed, and it has been established that, in terms of their ability to secrete luteinizing hormone (LH) in response to the specific secretogogue luteinizing-hormone-releasing hormone (LHRH), the cells become maximally responsive after 48 h. The attached cells also allow the short-term kinetics of LH secretion to be followed with great facility; and, when cells allowed to recover for 48 h are used, it is shown that in response to LHRH the pattern of LH release is biphasic.


Use of the avidin-biotin complex for the localization of actin and myosin with fluorescence microscopy. A new indirect method for fluorescence localization of proteins making use of the avidin-biotin complex is described. We have prepared both a biotin-modified rabbit heavy meromyosin (BHMM) and a biotin-modified antibody to a smooth muscle myosin. After fixation, cells can be treated with either BHMM, which binds to actin, or the biotinyl antibody, which binds to myosin. In a second step the cell are treated with a fluorescent derivative of avidin (Fl-avidin) which binds to the biotinyl proteins and thus indirectly reveals the location of the cellular action or myosin.


Light meromyosin paracrystal formation.

Light meromyosin paracrystal formation. STUDIES OF PARACRYSTAL FORMATION BY COLUMN PURIFIED LIGHT MEROMYOSIN (LMM) PREPARED IN A VARIETY OF WAYS LED TO THE FOLLOWING CONCLUSIONS: (a) different portions of the myosin rod may be coded for different stagger relationships. This was concluded from observations that paracrystals with different axial repeat periodicities could be obtained either with LMM framents of different lengths prepared with the same enzyme, or with LMM fragments of identical lengths but prepared with different enzymes. (b) Paracrystals with a 14-nm axial repeat periodicity are most likely formed by the aggregation of sheets with a 44-nm axial repeat within the sheets which are staggered by 14 nm. All of the axial repeat patterns expected from one sheet or aggregates of more than one sheet, on this basis, were observed in the same electron micrograph. (c) C-protein binding probably occurs preferentially to LMM molecules related in some specific way. This was concluded from the observation that the same axial repeat pattern was obtained in paracrystals formed from different LMM preparations in the presence of C-protein, regardless of differences in the axial repeat obtained in the absence of C-protein. (d) Nucleic acid is responsible for the 43-nm axial repeat patterns observed in paracrystals formed by the ethanol-resistant fraction of LMM. In the absence of nuclei acid, paracrystals with a 14nm axial repeat are obtained. (e) The 43-nm axial repeat pattern observed with the ethanol-resistant fraction of LMM is different for LMM preparations obtained by trypsin and papain digestions.


Immunofluorescence studies of neurofilaments in the rat and human peripheral and central nervous system. Localization of antisera to neurofilament antigens derived from rat peripheral nerve was carried out in tissues of rat and human peripheral and central nervous systems by indirect immunofluorescence. Unfixed and chloroform-methanol-fixed frozen sections of tissues were incubated in purified IgG of the experimental rabbit antisera and subsequently exposed to goat anti-rabbit IgG conjugated with fluorescein isothiocyanate. Control studies were conducted on identical tissue preparations incubated in the same concentrations of nonspecific rabbit IgG or in experimental rabbit IgG absorbed with extracts of rat peripheral nerve containing neurofilament antigen. Extensive immunofluorescence was observed in rat and human peripheral and central nervous systems. The distribution and configuration of immunofluorescence corresponded to neurofilament-rich structural components of these tissues. Prominent immunofluorescence was also noted in neuronal cell bodies of spinal sensory ganglia, especially in perikarya of the large neuronal type. Immunofluorescence of the central nervous system was located predominantly in myelinated axons of the white matter in cerebrum, cerebellum, brain stem, and spinal cord. Less intense immunofluorescence was also seen in neuronal perikarya and in short thin linear processes of grey matter.


Gas-liquid chromatography of fecal neutral steriods. A method is described for the analysis of fecal neutral steriods with a dual-column gas-liquid chromatography (GLC) system. After saponification of the fecal slurry, the neutral steroids were extracted with hexane. The GLC separation of the compounds and quantitation were achieved by simultaneous injection of the derivatized  and derivatized aliquots of the extract onto dual colmuns under identical conditions. The neutral steroids of interest were than identified by matching the  retention times with those of known standards, and identification was confirmed by use of an interfaced GLC high-resolution mass spectrometry system. The detection limit was 0.003 mg of steroid/g of fecal slurry. The pricision of the method is illustrated by a relative standard diviation of 2-10% and a recovery of neutral steroids from 73-96%. The method was applied to the determination of fecal neutral steroids in a "High protein diet in colon cancer study". A considerably larger level of coprostanone than of coprostanol was observed. Data on neutral steroids in fecal samples from subjects on different diets are the subject of a separate publication.


Further evaluation of the test for detection of antibody-coated bacteria in urine sediment. To assess the reproducibility of the test for detection of antibody-coated bacteria in urine sediment as it might be used in a diagnostic laboratory for classification of urinary tract infections, multiple urine specimens from 83 patients were tested. The results were reproducible and consistent, or if inconsistent potentially explainable, in all but four patients. The explanation for inconsistencies include the immune response to the infecting bacteria, nonspecificity of the antibody coating the bacteria, antibody in prostatic secretions, and antibody-coated bacteria contaminating the urine specimens.


Template method for the fluorescent-antibody technique. An easily constructed and inexpensive template has been developed, which enables the fluorescent-antibody technique to be applied to serial dilutions and allows 18 assays on a single microscope slide.


Comparison of the BACTOMETER microbial monitoring system with conventional methods for detection of microorganisms in urine specimens. A rapid impedance-based screen for bacteriuria (less than 10(5) organisms per ml) was compared with a standard loop method, using 156 clinical urine specimens. Agreement between the methods was 96.8%.


Glucagon and insulin binding to liver membranes in a partially nephrectomized uremic rat model. To investigate the role of glucagon and insulin receptor binding in the glucagon hypersensitivity and insulin resistance which characterize the glucose intolerance of uremia, liver plasma membranes were prepared from control rats (blood urea nitrogen [BUN] 15+/-1 mg/100 ml, creatinine 0.7+/-0.2 mg/100 ml), and from 70% nephrectomized rats (BUN 30+/-2 mg/100 ml, creatinine 2.2+/-0.2 mg/100 ml), and from 90% nephrectomized rats (BUN 46+/-3 mg/100 ml, creatinine 4.20+/-0.7 mg/100 ml), 4 wk after surgery. As compared to controls, the 90% nephrectomized rats had significantly higher levels of plasma glucose (95+/-4 vs. 125+/-11 mg/100 ml), plasma insulin (28+/-9 vs. 52+/-11 muU/ml), and plasma glucagon (28+/-5 vs. 215+/-18 pg/ml). Similar, but less marked, elevations were observed in the 70% nephrectomized animals. In liver plasma membranes from nephrectomized rats, specific binding of (125)I-glucagon was increased by 80-120%. Furthermore, glucagon (2 muM)-stimulated adenylate cyclase activity in nephrectomized rats was twofold higher than in controls. In contrast, fluoridestimulated adenylate cyclase activity was similar in both groups of rats. In marked contrast to glucagon binding, specific binding of (125)I-insulin to liver membranes from nephrectomized rats was reduced by 40-50% as compared to controls. Data analysis suggested that the changes in both glucagon and insulin binding are a consequence of alterations in binding capacity rather than changes in affinity. Liver plasma membranes from nephrectomized rats degraded (125)I-glucagon and (125)I-insulin to the same extent as control rats. THESE RESULTS DEMONSTRATE THAT: (a) the 70 and 90% nephrectomized rats simulate the hyperglycemia, hyperinsulinemia, and hyperglucagonemia observed in clinical uremia; (b) in these animals specific binding of glucagon to liver membranes is increased and is accompanied by higher glucagon-stimulated adenylate cyclase activity; and (c) specific binding of insulin is markedly decreased. These findings thus provide evidence of oppositely directed, simultaneous changes in glucagon and insulin receptor binding in partially nephrectomized rats. Such changes may account for the hypersensitivity to glucagon and may contribute to resistance to insulin observed in the glucose intolerance of uremia.


Corticosteroid effect on granulopoiesis in mice after cyclophosphamide. Corticosteroids cause an enhanced return of granulopoiesis as measured by in vitro growth of granulocytic progenitor cells (CFU-C) in mice treated with cyclophosphamide. After methylprednisolone and cyclophosphamide, a greater than threefold increase in marrow CFU-C was measured on day 4 compared to mice given cyclophosphamide alone (29,700+/-200 vs. 8,400+/-700/humerus). The accelerated return of marrow CFU-C was observed with cyclophosphamide in doses of 200 and 450 mg/kg and methylprednisolone, 2-20 mg/kg, with no significant differences using >5 mg/kg, and was detected when dexamethasone was used in place of methylprednisolone. This effect was accompanied by similarly enhanced splenic granulopoiesis as measured by CFU-C concentration. Levels of colony stimulating activity did not differ in mice given methylprednisolone and cyclophosphamide or cyclophosphamide alone. Corticosteroids appear to enhance the return of CFU-C by altering the proliferative state of granulocytic progenitor cells. CFU-C survival to in vitro (3)HTdR suicide increased from 72+/-4% on day 1 after cyclophosphamide to 90+/-6% in animals given both cyclophosphamide and methylprednisolone. Increased survival after (3)HTdR suicide was also observed when methylprednisolone alone was given. After treatment with cyclophosphamide and methylprednisolone, blood neutrophils increased more rapidly and improved survival to infection with Candida albicans was observed. These studies demonstrate that corticosteroids have a beneficial effect on marrow regeneration after myelotoxic chemotherapy with cyclophosphamide and suggest that they act by altering cell cycle characteristics of granulocyte progenitor cells.


The mobilization of arachidonic acid in platelets exposed to thrombin or ionophore A23187. Effects of adenosine triphosphate deprivation. In studies conducted with human gel-filtered platelets, we have found: (a) that the release of serotonin and transfer of [3H]arachidonic acid from phosphatidylcholine and phosphatidylinositol to plasmalogen phosphatidylethanolamine which are associated with  the activation of platelets by thrombin are both strongly dependent upon the presence of metabolic ATP; (b) that serotonin release and arachidonic acid mobilization in labeled phosphatides are promoted by the calcium ionophore A-23187 in media free of calcium ions; (c) that inhibitors of ATP synthesis, while leading to impairment of the release reaction induced by ionophore, do not inhibit ionophore-stimulated mobilization of arachidonic acid. We conclude that the activation of phospholipase A2 responsible for freeing arachidonic acid from platelet phosphatides is solely dependent upon the increased cytoplasmic levels of calcium ions promoted by either ionophore or, in an energy-dependent fashion by thrombin. Phospholipase activation is not a function of latent hydrolytic activity made available by the release reaction.


Factors affecting the value of a simple biochemical scheme for identifying Enterobacteriaceae: the reproducible recognition of biotypes. A biochemical screening scheme on agar media for differentiating Enterobacteriaceae in a hospital laboratory is evaluated. Careful observation of test reactions within the scheme permitted the recognition of 78 biotypic reaction patterns which could contribute to epidemiological surveillance. The limitations of the technique are described and discussed and methods of importance in ensuring the reproducibility of reaction patterns emphasised.


The API ZYM system in the identification of Gram-negative anaerobes. The API ZYM reactions of type species of Gram-negative anaerobes representative of those encountered in human infections and of 56 clinical isolates of such organisms, identified by conventional techniques, were investigated. The API ZYM test clearly distinguished between the different genera and species examined and appears to provide a simple, reliable method for the identification of this group of organisms.


Modification of the microimmunofluorescence test to provide a routine serodiagnostic test for chlamydial infection. A modification of the microimmunofluorescence test to provide a practicable routine serodiagnostic test for detecting and characterising chlamydial infection is described which uses four antigen pools, one of which corresponds with each of the four main clinical and epidemiological types of chlamydial infection. The three subgroup A Chlamydia (Chlamydia trachomatis) pools are: pool 1, hyperendemic trachoma TRIC agent serotypes A, B, and C; pool 2, paratrachoma TRIC agent serotypes D, E, F, G, H, I, and K; pool 3, lymphogranuloma venereum (LGV) agent serotypes L1, L2, and L3. Pool 4 contained four representative isolates of subgroup B Chlamydia (Chlamydia psittaci). For routine purposes sera need be screened only against these four representative antigen pools. This will detect antibody and indicate which clinical and epidemiological type of chlamydial infection is implicated, thereby clearly distinguishing those infections that are due to C. psittaci. The pattern of the cross-reactions may indicate the individual serotype involved, and further titration requiring a maximum of four individual antigens is sufficient to determine the serotype. The slight loss in sensitivity (twofold) is more than compensated for by the reduction in cost and the tenfold increase in the total number of sera which can be examined.


Laboratory assessment of physical and chemical methods of preserving urine specimens. Three methods of preserving simulated specimens of urine were studied with six test strains of bacteria. Viable counts were measured by a surface viable count and by the filter-paper-strip method during a holding period of 72 hours. Refrigeration at approximately 4 degrees C was effective and reliable. Boric acid (1-8%) at room temperature was toxic for the strain of Escherichia coli at a density of 10(7) cfu/ml but this may not be significant at the higher concentration of bacterial cells often found in clinical specimens. NaCl-polyvinylpyrrolidone (PVP) solutions containing PVP of mol. wt 44 000 or 700 000 were not effective; they were toxic for the Gram-negative strains and did not retard the growth of Micrococcus subgroup 3. The two methods of measuring viable counts were compared for specimens held under different conditions; the specificity of the filter-paper-strip method was high but the sensitivity was low when many of the specimens contained approximately 10(5) cfu/ml.


Intracellular lysozyme and lactoferrin in myeloproliferative disorders. Samples from 49 cases of myeloproliferative diseases were tested by an immunocytochemical technique for leucocyte lysozyme and lactoferrin. The presence of these constituents in myeloid precursors from cases of acute and chronic myeloid leukaemia reflected the degree of cellular maturation, lysozyme appearing (as it does in normal myeloid cells) at the stage of primary granule production (in promyelocytes), while lactoferrin wad detectable only in more mature, secondary granule-containing myeloid cells. Auer rods stained positively for lysozyme, in keeping with their relationship to primary granules. Monocytes from five cases of leukaemia showing predominantly monocytic differentiation were indistinguishable from normal monocytes in their staining reactions for lysozyme despite the presence of raised serum and urinary lysozyme levels. In four cases of acute myeloid leukaemia circulating polymorphs deficient in lactoferrin were detected: in one of these cases a similar percentage of polymorphs was lysozyme negative.


Immunohistochemical study of lichen planus. The distribution of IgM and IgG in 20 cases of lichen planus and five cases of non-specific inflammation were studied by the unlabelled antibody-enzyme (PAP) technique. Routine paraffin sections were used. In lichen planus deposits of immunoglobulins were seen in and around epithelial cells, in colloid bodies, at epidermo-dermal junctions, and in some inflammatory cells. All cases examined for IgM and eight out of 13 cases examined for IgG were positive. The peripheral migrating epidermal cells were mostly negative. The application of the PAP technique to skin biopsy and the significance of the findings in lichen planus are discussed.


Cell volume to aid analysis and technique of somatic cell counts in milk. In conjunction with a Coulter Counter, somatic cells in milk were sized by electronic analysis. Quarter milk from cows with mastitis had a cell volume peak with a modal cell volume of 102  mu3 while milk from healthy quarters had no peak. Bulk milks with a peak had higher cell counts than milks where there was no peak. Dimensions of peak from bulk milks were the same as from quarter milks. Modal cell volume and cell count varied after milks were fixed with varying concentrations and types of fixative. The concentration of fixative recommended by the International Dairy Federation was sufficient, but only marginally so, and it should be increased.


Cerebral ventricle cannulation in the calf. A stereotaxic apparatus and procedure for surgically implanting guide cannulae for access to the cerebral ventricles of the calf are described. This type of implant permits chronic ventricular injections and collections of mililiter quantities of cerebrospinal fluid from the nonanesthetized calf.


Pinch grafting for chronic ulcers on lower extremities. Chronic ulceration of the skin of the lower extremities continues to be a difficult therapeutic problem. Pinch grafting of ulcers of the legs from a variety of causes, has proved to be highly successful. The success rate and complications of this procedure performed in the Department of Dermatology at the University of Iowa over a five-year period (1971-1975) are reviewed. The basic technique and pre-operative and post-operative care are outlined. Management of difficult cases by using such techniques as the Jobst pump, heterograft skin dressings, and sublesional steroid injection is also discussed.


Complications of the tunnel-graft procedure for attachment of a hairpiece: a review of 100 cases. The office procedure of creating tunnel grafts for attachment of hairpieces is not without complications. Early complications such as a hematoma under a graft, bacterial infection, and maceration of the epithelium lining the tunnels tended to be acute, but minor. Late complications, such as cutting in of the clips into the roofs of tunnels or stretching of the tunnel-graft roof, were mainly of a mechanical nature. Recommendations for reducing the indicence of such complications are made.


A double-blind evaluation of the use of nebulized metaproterenol and isoproterenol in hospitalized asthmatic children and adolescents. This study compared two beta-adrenergic agents in the treatment of severe asthma of children and adolescents. Thirty patients admitted to Children's Orthopedic Hospital and Medical Center with acute asthma were treated with either isoproterenol 0.05% (iso) or metaproterenol 0.5% (meta) nebulized in physiologic saline and delivered with O2 in addition to intravenous hydration, aminophylline, and corticosteroids. Vital signs, blood gases, and pulmonary function were monitored frequently. No adverse reactions to the study drugs were encountered. Pulse rates increased similarly after treatments with both drugs. Patients treated with meta had somewhat higher rates than those treated with iso. Diastolic blood pressure decreased significantly from admission in the meta group compared to the iso group. For FVC and FEV1, the meta group had greater percentage increases from admission than the iso group at all times, with the difference being significant (p less than 0.05) at 1 and 12 hr. The same trend occurred for FEF 25%-75%. Though iso seemed to cause greater increases in flow immediately after administrations, these changes were transient compared to improvement caused by meta. Thus, meta seemed to be more effective than iso in reversing bronchospasm as measured by certain pulmonary function parameters.


Psychic reactions to long-term hemodialysis: evaluation in the elderly by means of the Draw-A-Person Test. Twelve patients who attended a daytime session in a hemodialysis unit were studied. Most were older than patients whose adaptation has been the subject of previous reports. Problems of life satisfaction for these elderly patients often are different from those for younger persons. Most of the group showed maladaptive reactions which interfered with their activities when away from the unit. The Draw-A-Person test was a useful tool in pinpointing these reactions, which can lead to feelings of helplessness and depression. Factors which encourage good adaptation to long-term hemodialysis are discussed.


The role of activated accessory cells in preventing immunosuppression by hydrocortisone. The generation of humoral immunity in vitro by normal and antigen-primed mouse spleen cells was suppressed by in vitro treatment with hydrocortisone. Functions of normal and antigen-activated helper T lymphocytes and of accessory cells were inhibited by the corticosteroids. Spleen cells cultured overnight in medium containing fetal bovine serum became highly resistant to the effects of hydrocortisone. Similar resistance was found to occur when spleen cells were cultured with accessory cells that previously had been activated with bacterial lipopolysaccharide. These studies show that immunologically nonspecific processes significantly alter the effects of the steroids on specific immune responses and suggest that accessory cell products modulate T cells in ways which differ from antigen induction.


Human monocyte-lymphocyte interaction: a new technique. A rosette-type assay of the physical interaction between lymphocytes and monocytes after treatment with neuraminidase-galactose oxidase (NGAO) is reported. Monocyte-lymphocyte (ML) rosette formation and subsequent lymphocyte proliferation occurred when either lymphocytes or homologous monocytes were treated with NGAO and cultured together. Maximal ML rosette formation took place at 37 degrees C 4 hr after culture in media containing 10% serum at lymphocyte to monocyte ratios of 10:1 to 20:1. The percentage of rosette formation correlated with the extent of thymidine incorporation when increasing concentrations of NGAO were used. When NGAO-treated monocytes were added to untreated T and non-T lymphocytes, they bound preferentially to T lymphocytes and induced proliferation only in the T subpopulation. These results indicate that the ML rosette assay measures a highly specific monocyte-lymphocyte physical interaction after a mitogenic stimulus which is an early event in lymphocyte activation since it reflects the degree of subsequent lymphocyte proliferation.


Erythrocyte membrane-associated immunoglobulins during malaria infection of mice. Immunoglobulin (Ig) is associated with erythrocyte membranes during infection of A/J mice with Plasmodium berghei. It was detected by agglutination of the cells with rabbit antibodies to mouse IgG and by a radioimmunoassay with 125I-labelled rabbit antibodies to mouse IgG. As shown by the degree of agglutination and of binding of radiolabeled antibodies to the erythrocyte membranes, the amount of Ig increased with time after infection and paralleled parasitemia and reticulocytosis. The erythrocyte-associated immunoglobulins are mainly IgG but IgM was also present on the cells of some mice. A large proportion of the Ig could be eluted at 37 degrees C and was analyzed by immunoprecipitation and acrylamide gel electrophoresis. A radioautographic study with 125I-labeled anti-mouse IgG revealed that both parasitized and nonparasitized reticulocytes of infected mice had much larger amounts of membrane-bound immunoglobulin than did mature, nonparasitized erythrocytes. The nature of the bonds between the Ig and the surface membrane of the reticulocytes is not known. The Ig could be part of immune complexes nonspecifically bound to the cell surface. However, since we have not detected Fc or C3d receptors on reticulocytes, it is possible that the Ig constitutes autoantibodies against reticulocytes or antibodies against parasitic antigens present on the cell membrane.


In vivo immune response suppression by the supernatant from concanavalin A-activated spleen cells. Supernatants from concanavalin A- (Con A) activated murine spleen cells have been shown to suppress the in vitro plaque-forming cell (PFC) response to sheep red blood cells (SRBC). The present study examined the effect of such Con A-activated spleen cell supernatants (herein termed CONS) on the in vivo immune response to SRBC in C57BL/6, BALB/c and CDF1 mice. CONS derived from BALB/c spleen cells suppressed direct PFC 4 and 8 days after immunization with 2 X 10(8) SRBC. CONS also suppressed indirect PFC 8 days after immunization, as well as serum hemagglutinins to SRBC. The PFC response of C57BL/6 (H-2b) mice was suppressed as much as that of BALB/c (H-2d) by CONS derived from BALB/c mice, indicating a lack of H-2 specificity of the CONS. In addition to suppression of the antibody response to SRBC, in vivo CONS administration resulted in reduction in spleen cell number. This reduction was not sufficient to explain the decreased PFC response. When the CONS was separated into less than 10,000 m.w. and greater than or equal to 10,000 m.w. fractions, the immunosuppressive activity was found in the less than 10,000 m.w. fraction. This observation suggests that intact interferon, SIRS, and MIF were not responsible for the results obtained.


Identification of RNA as a complement inhibitory component in an extract of Ehrlich ascites tumor cells. A factor capable of inhibiting complement was obtained from intact Ehrlich ascites tumor cells by mild extraction with phosphate-buffered saline (PBS). The inhibitor caused a decrease in extent of lysis of EAC14 with a concomitant extension of Tmax. EA, EAC1, EAC4 and EAC142 were all less susceptible to complement-mediated lysis after treatment with the tumor cell extract. Partial purification of a complement inhibitor was accomplished. The inhibitor was rich in RNA and its activity was totally destroyed by RNAase but not DNAase. RNA from mouse tissues, yeast, and Escherichia coli also inhibited complement hemolytic activity. The partially purified material only inhibited lysis of EAC1 and EAC14. Slow inhibition of fluid phase C1 was also demonstrated. In addition, RNA-rich partially purified tumor cell extract was capable of precipitating with purified human C1q.


Radioimmunoassay of IgG and IgM rheumatoid factors reacting with human IgG. Although IgG rheumatoid factor may play a central role in the pathogenesis of rheumatoid arthritis, previously there have been no precise methods for its specific measurement in serum and synovial fluid. This paper describes a solid phase radioimmunoassay for the independent quantification of IgM and IgG rheumatoid factor reacting with the Fc fragment of human IgG. As measured by this assay, serum IgG rheumatoid factor levels differed significantly between patients with seropositive and seronegative rheumatoid arthritis and normal control subjects. In addition, several sera and joint fluids from patients with seropositive rheumatoid arthritis, even without vasculitis, were shown by gel chromatography to have acid-dissociable complexes of IgG rheumatoid factor suggestive of IgG-IgG dimer or trimer formation.


Fc receptors on mouse effector cells mediating natural cytotoxicity against tumor cells. Mouse effector cells mediating natural cytotoxicity against tumor cells have been previously thought to be lymphocytes that lack any detectable cell surface markers. The present study presents evidence for receptors for the Fc portion of IgG on these cells. By adsorption of cytotoxic spleen cells on monolayers of sheep erythrocytes (E) plus IgG antibodies to sheep erythrocytes (EA), 50 to 96% of the total cytotoxic reactivity could be removed. Parallel adsorption of cells on E monolayers or on EA monolayers coated with protein A, to block the Fc portion of IgG, resulted in little or no depletion of cytotoxic activity. The presence of Fc receptors on the NK cells was confirmed by combining EA rosette formation with velocity sedimentation at unit gravity. Peak cytotoxicity occurred at the same sedimentation velocity as the peak of Fc-positive cells. After EA rosette formation, there was a shift to a higher sedimentation velocity in the Fc-positive cells and in the natural cytotoxic activity. The increase in sedimentation velocity of NK activity that was observed in these experiments indicated that most of the cells had only bound a small number (three or four) of antibody-coated erythrocytes. Together, these data indicate that cells with Fc receptors account for most of the total lytic activity of normal mouse spleen cells.


Strain variation in BCG-induced chronic pulmonary inflammation in mice. I. Basic model and possible genetic control by non-H-2 genes. C57BL/6 mice (haplotype H-2b) responded in a dose-dependent fashion to killed BCG by marked enlargement of the spleen and lung. Neither CBA nor C3H mice (haplotype H-2k) responded to such treatment. Pulmonary inflammation in responder B6 animals was characterized by a marked chronic interstitial and alveolar granulomatous process, and was accompanied by occasional granulomata, hyperemia, and loss of architecture in the spleen. Inflammation in non-responder CBA and C3H animals was minimal in both the lung and spleen. The response does not appear to be controlled by genes within the major histocompatibility complex, but is associated with a C57 background. B10.BR mice (responder background, H-2k) were responder animals and C3H.SW mice (nonresponder background, H-2b) were nonresponders. In addition, all animals tested with a C57 background were responders even though two of these strains were not H-2b (C57BL/Ks, H-2d and C57Br/cd, H-2k). The resolution of the mechanism of genetic control of this response in mice may provide information relevant to possible genetic control of chronic pulmonary inflammation in man.


No direct association between Ia antigens and Fc receptors on the B cell membrane. The capping of Ia antigens does not induce redistribution of Fc receptors (FcR) on B lymphocytes. This rules out the possibility of a unidirectional association between Ia and FcR such as has been reported to link Ig and FcR. Ia-capping was achieved with hapten-sandwich antibodies devoid of Fc regions: hapten-conjugated Fab anti-Ia followed by (Fab')2 anti-hapten antibody. Three different immune complex systems were used to label FcR. With fluorescent double labeling, Ia and FcR were readily distinguished. The independent labeling and surface mobility of Ia and FcR are considered in connection with reports of the inhibition of FcR by anti-Ia antibodies.


Release of phospholipids from complement-mediated lesions on the surface structure of Escherichia coli. When varying numbers of sensitized, 14C-labeled bacteria were treated with a certain amount of complement, in a fixed reaction volume, 14C compounds were liberated into the surrounding medium in proportion to the number of the bacteria, whereas the amount of the phospholipids liberated was constant regardless of the number of  the bacteria even in the range of relative excess of complement. Since it is conceivable that a certain amount of complement might form a fixed number of lesions on the surface of all the sensitized bacteria, the amount of the liberated phospholipids seems to be proportional to the number of complement lesions. The 14C-materials released from complement-attacked bacteria were analyzed by isopycnic sucrose density gradient ultracentrifugation and they were mainly free phospholipids and other smaller molecules. A small amount of the smaller membrane proteins were also released as revealed by acid and SDS-polyacrylamide gel electrophoresis. These results suggest that the release of phospholipids is due to the displacement of membrane lipids by the complexes of the late acting complement components during their insertion into the membrane lipid bilayer.


Phospholipase C-labeled anti-human IgG: inhibition by human IgF. We report here the inhibition of enzyme activity of phospholipase C-anti-human rabbit IgG conjugate complexed with human IgG. Phospholipase C activity was measured by assaying the release of hemoglobin from erythrocytes. The degree of inhibition varies depending on the relative amount of IgG utilized. A water soluble carbodiimide linking yields a conjugate which is inhibited by IgG, while conjugate prepared by glutaraldehyde is not. Immunodiagnostic assays requiring detection of low levels of antigen or antibody may be possible utilizing the phospholipase C-erythrocyte technique.


Problems with radiolabelling egg albumin for the Farr ammonium sulphate technique for antibody estimation. High concentrations of passive haemagglutinating antibodies were raised in sheep immunised with egg albumin (Ea) in Feund's adjuvant. Further attempts to study the antigen binding capacities and avidities of these antisera by the Farr ammonium sulphate technique were hindered by unsuccessful attempts to label Ea with 125Iodine by either the chloramine T or sodium nitrite-HCl methods. This problem was attributed to the unavailability of tyrosine in the native protein. This was solved by labelling Ea with the Bolton-Hunter reagent which does not require the presence of tyrosine. However labelled Ea binds non-specifically to normal serum to give high control values of circa 40% binding. Despite this it was possible to estimate antibody concentrations and avidities of lamb sera.


Delayed hypersensitivity in the hamster cheek pouch. A method is described for eliciting a delayed hypersensitive cell mediated immune response in the Syrian hamster cheek pouch. Using a sensitizing dose of 0.1% DNCB and a challenge dose of 0.01% DNCB 14 days later resulted in a histologically positive lymphocyte infiltrate in the challenge area. These results indicate that the hamster cheek pouch may not be an immunologically inert site and may be used for cell mediated immunity studies.


The statistical distribution of macrophage migration distance and its application to MIF test. The measurement of the distance of macrophage migration is very simple and reasonable as a technique in MIF assay. By the use of this technique, a large scale model experiment was performed and its data were analysed statistically. As the result of analysis, Student's t-test for significance of the difference between control and test readings of migration distance at low activity level in the MIF test was justified, and it was proved that it is applicable to the routine MIF test. In addition to this, the measurement of migration distance was compared with that of migration area.


Some methodologic aspects of the chromium chloride method for coupling antigen to erythrocytes. The influence of various methodological variables on the CrCl3 technique for coupling antigen to the surface of erythrocytes has been investigated. Tests should be performed in protein- and phosphate-free medium. The CrCl3 stock solution should be stored at a pH of 5.0 and working dilultions prepared in acetate buffer (pH 5.5). The coupling procedure itself was performed as described by Goding (1976) with slight modifications. The relation between antigen and CrCl3 concentrations was found to be of crucial importance as excess of antigen inactivates CrCl3 whereas lack of antigen or excess of CrCl3, leads to spontaneous agglutination. A preliminary test based on determination of the minimum concentration of CrCl3 which produces spontaneous agglutination in the absence of antigen, and coupling if antigen is available, can be employed to predict the optimal concentration with sheep, human or chicken red blood cells. The pH-dependency of the coupling process is emphasized.


The fluorescence-plaque test--a new sensitive technique for the detection of antigens on antibody plaque-forming cells. A new technique is described which allows the detection of small amounts of antigen on the membrane of antibody plaque-forming cells. The test system combines the indirect immunofluorescence test and the Jerne plaque technique, and is about 10 times more sensitive than the ordinary cytotoxic plaque technique.


A microassay for leukocyte migration: analysis of its reproducibility. A reliable microassay for human leukocyte migration is described by which 50 to 100 assays can be performed each day in quadruplicate with the number of indicator cells obtainable from 10 to 20 ml of peripheral blood. The reproducibility of this method is demonstrated with respect to the variability among replicate test wells (including reading), the variability among different test readers, the variability among replicate cultures, and the variability of using indicator cells from different subjects. A microculture system is described that requires only 75,000 mononuclear cells to consistently produce detectable polymorphonuclear leukocyte migration inhibition factor in response to PPD. The reproducibility of this culture system is demonstrated with respect to the variability of lymphocyte responsiveness on repetitive testing in the same individuals.


A rapid method for reading migration inhibition tests. This article describes how the extent of cell-migration can be read directly using the Field iris of a microscope.


The avidity of IgM antibody in high and low responder rats. This study examined IgM antibody produced by highly responding ACI and poorly responding F344 rats follwing immunization with poly(Glu52Lys33Tyr15) or poly(Glu52Lys33Tyr15) aggregated with methylated bovine serum albunim (MeBSA). The ACI rats produced both IgM and IgG plaque-forming cells (PFC) following immunization with either form of antigen. The F344 rats did not respond to unaggregated poly(Glu52Lys33Tyr15), but they produced significant amounts of IgG PFC and extremely small amounts of IgM PFC after immunization with poly(Glu52Lys33Tyr15)/MeBSA. Both high and low responder rats had similar kinetic profiles of IgM antibody production, and this antibody had nearly identical avidity in both strains with no evidence for any maturation in avidity. thus, one of the genetic defects in the antibody response to poly(Glu52Lys33Tyr15) is an inability of the F344 strain to produce large amounts of IgM in response to this antigen.


Cutaneous stimulus registration and information processing in the somesthetic cortex. The author reviews the evidence that the acquisition of sequential tactile information and stereometric object identification by manual exploration is a process which is analogous to the scanning of visual scenes. The hypothesis that somatosensory and visual maps in the respective primary cortical projection areas are of the same analytical structure is argued. New experimental data documented in subhuman primates demonstrate that neural responses to cutaneous stimuli depend on situational factors and on behavior-controlling instructions conveyed over another sense modality and that as a result the redundancy of cutaneous stimulus encoding in the somesthetic cortex is reduced.


Neural-epithelial interactions in sensory receptors. The specific types of relationships that exist between sensory nerve fibers and the associated epithelial cells have been reviewed in a variety of sensory systems. In many sensory systems, perhaps most, specificity of terminal epithelial element provides a degree of specificity to the cytologic pattern of the terminal receptor. We can see examples of such interaction, not only in the case of intraepithelial nerve fibers where nerves tend to be associated with Merkel cells, but also in the case of corpuscular receptors, both of these being examples of mechanosensory systems. Equivalent types of specificity are present within the special senses, where a specific receptor cell can be described. These findings strongly suggest that specificity in the peripheral nervous system is the rule rather than the exception, a thesis proposed over a hundred years ago by F. Merkel.


Electron microscopy of cutaneous nerves and receptors. Recent ultrastructural observations on the connective tissue sheaths of nerves, Schwann cell-axonal relations, and nerve terminals and receptors are reviewed. It seems likely that endoneurial collagen is formed by perineurial cells during development and postnatally. New observations on "collagen pockets" are presented. Attention is drawn to freeze-fracture studies of peripheral nerve, particularly in relation to junctional complexes associated with compact myelin, and further application of the technique is considered. Current views on Merkel cells, encapsulated endings, and free nerve terminals are discussed.


Mechanisms of the flare reaction in human skin. The diffuse area of arteriolar vasodilation surrounding a region of recently injured human skin (axon reflex flare) is dependent upon the integrity of nerve fibers with cell bodies located in dorsal root ganglia. Evidence is presented to indicate that a vasodilator peptide similar to a kinin, neurotensin, or substance P, is implicated in the chain of biochemical events responsible for the transient shift in vascular tonus observable as the flare reaction.


The detection of platelet isoantibodies by membrane immunofluorescence. A membrane ummunofluorescence test for the detection of platelet isoantibodies is described. Gel filtration of the incubation mixture was incorporated in the procedure and proved effective for the removal of serum proteins from the platelet suspension. With this technique isoantibodies were found in the serum of 13 out of a group of 16 patients who had received multiple transfusions. The results were checked by measuring the uptake of 125I-labeled anti-IgG fraction by gel-filtered platelets. Subsequently the membrane immunofluorescence method was also compared with established techniques described for the detection of isoantibodies such as the microtest for lymphocytotoxicity and a complement-fixation method and the procedures based on the release of labeled serotonin, the phagocytosis of chromium-tagged platelets, the increase of platelet factor 3 activity, and on platelet aggregation. We had the opportunity to investigate the serum of one patient for the presence of isoantibodies against platelets from HLA identical siblings both before and after the administration of their platelets. On the basis of this experience it is concluded that the membrane immunofluorescence test for platelet isoantibodies is a relatively simple method with a high degree of specificity and adequate sensitivity.


Cancer of the floor of the mouth: surgical management. After discussing the changing incidence and known difficulties in the management of mouth floor cancer, reasons are given for the increased use of surgery in treating the more extensive lesions. The significance of the topographic anatomy, patterns of lymph drainage and surgical pathology is emphasized. Principles of treatment, indications and types of operation are discussed for the main lesions encountered. Following this the preparation of patients for surgery, the methods of anaesthesia and certain technical details of surgical treatment are given. The importance of after-care and particular measures to avoid complications are stressed and conclusions reached. The paper is based on experience in the Head and Neck Unit of the Royal Marsden Hospital, London, mainly since 1965. It also derives from an extensive report on 189 cases of floor of mouth cancer seen and treated at the Hospital from 1950-1974.


Dose response to fenoprofen in an antipyretic study of fenoprofen and propoxyphene. Single oral doses of 0, 50, 100, and 200 mg (acid equivalents) of fenoprofen calcium provide essentially linear increases in antipyretic activity over a six-hour period in patients with fever due to acute upper respiratory tract infection. During this same time interval, 200 and 400 mg doses apparently had equal efficacy. Single oral doses of 200 and 400 mg propoxyphene napsylate had no significant effect on the fever of patients with acute respiratory tract infections. No significant interaction between propoxyphene and fenoprofen, related to the antipyretic effect of fenoprofen, was demonstrated.


A multicenter analgesic study using single doses of placebo, propoxyphene and acetaminophen. Six studies were conducted using identical protocols. The design involved single oral doses of placebo, propoxyphene, acetaminophen and a combination of the two drugs. Postpartum patients with either postepisiotomy or uterine cramp pain participated. Both efficacy and adverse reports were evaluated. The differences among medications were consistently less than one standard deviation from the mean for the studies indicating a lack of sensitivity to drug effects. In order of increasing effectiveness based on the pooled data, the medications were placebo, propoxyphene, acetaminophen, and the combination.


Oestradiol-17beta and pituitary responsiveness to luteinizing hormone releasing factor in the rat: a study using rectangular pulses of oestradiol-17beta monitored by non-chromatographic radioimmunoassay. Plasma concentrations of oestradiol-17beta were measured by a non-chromatographic radioimmunoassay during the oestrous cycle, after the s.c. injection of 2-5 or 10 microgram oestradiol benzoate (OB), or the s.c. implantation of Silastic capsules containing crystalline oestradiol-17beta. The profile of endogenous plasma oestradiol-17beta concentrations was similar to that reported by other workers, and lay between the concentrations produced by the low and high doses of OB. The rectangular pulses of increased plasma oestradiol concentrations, produced during the period of implantation of the Silastic capsules, were used to determine the time taken for oestradiol-17beta to exert its facilitatory effect on the gonadotrophin response to LH-releasing factor (RF). In animals ovariectomized at dioestrus, oestradiol, at concentrations similar to those reached during the peak of the spontaneous surge, first reduced the LH response. However, after 7 h, responsiveness increased significantly to reach a peak at 12 h. The FSH response was also greatest 12 h after ovariectomy. In animals ovariectomized at metoestrus the effect of oestradiol on the LH response was significantly less than in rats ovariectomized at dioestrus, and the FSH responses were lower than those in animals bearing empty capsules and examined at the same time after ovariectomy. These findings together with the effects of long-term exposure to sodium pentobarbitone are considered with respect to the possible mechanisms, including the priming effect of LH-RF, which may produce increased pituitary responsiveness after ovariectomy and exposure to oestrogen.


Effect of oestradiol-17beta exposure on the spontaneous secretion of gonadotrophins in chronically gonadectomized rats. The effect of oestradiol-17beta (administered in Silastic capsules) on gonadotrophin secretion in long-term gonadectomized rats has been investigated. In female rats, a daily afternoon surge of LH occurred which could be blocked by administering sodium pentobarbitone at 13.00 h. This, together with the fact that there was no significant difference between the LH-response to LH-releasing factor in the morning compared with the afternoon, supports the view that the LH surges are due to a neural rhythm. There was no significant diurnal variation in plasma FSH concentration, but an apparent diurnal variation was disclosed by administering sodium pentobarbitone on alternate days. There was no diurnal variation in plasma gonadotrophin concentrations in male animals implanted with an oestradiol capsule, or in animals of either sex bearing empty capsules.


An homologous radioimmunoassay for chicken follicle-stimulating hormone: observations on the ovulatory cycle. A highly purified FSH preparation has been used to develop a specific homologous radioimmunoassay for chicken FSH which is sufficiently sensitive and precise to measure the hormone in small sample (10-100 microliter) of plasma. The assay was used to measure plasma FSH in the chicken and turkey. The FSH concentration was higher in sexually mature chickens than in juvenile birds and further elevated after castration or ovariectomy. In turkeys, it was lower in birds held on a short daily photoperiod than in birds held on a long daily photoperiod. FSH rose in sexually quiescent female turkeys after injection of synthetic LH-releasing hormone and was increased in laying hens after injection of progesterone. No major changes were observed in FSH concentration during the chicken ovulatory cycle, although there was a small increase 15 and 14 h before ovulation.


Changes in pituitary responsiveness to luteinizing hormone releasing hormone in rams exposed to artificial photoperiods. Six adult Soay rams were subjected to an artificial lighting régime of alternating 4 month periods of long days (16 h light:8 h darkness) and short days (8 h light:16 h darkness) which induced a seasonal cycle in testicular development and regression during a period of 32 weeks. On 15 occasions during the study pituitary responsiveness was assessed by measuring the concentration of luteinizing hormone (LH) in jugular blood samples collected at frequent intervals after the intravenous injection of 1 microgram synthetic luteinizing hormone releasing hormone (LH-RH). It was shown that both the magnitude and duration of the induced release of LH changed in relation to the photoperiod; the magnitude was greatest during long days when the rams were sexually quiescent, while the duration of the LH release was greatest during short days at the peak of testicular activity. This pattern of responsiveness was modified by implantation of the rams with testosterone.


In-vitro studies of the effects of oestrogen pretreatment on the sensitivity of the immature female rat pituitary gland to stimulation with gonadotrophin releasing hormone. The effects of oestrogen priming on the sensitivity of the anterior pituitary gland to stimulation with gonadotrophin releasing hormone (GnRH) was investigated in immature female rats using a new organ culture technique. Hemipituitary glands obtained from animals primed with a single dose of oestradiol benzoate (OB; 20 microgram/100 g body weight) released significantly more LH when pulsed with GnRH (4 nmol/1) than did control hemipituitary glands. This potentiating effect was detectable as early as 5 days after birth. After a second stimulation, LH secretion remained high. These results were compared with those obtained from animals treated to induce increased levels of endogenous oestrogen on day 26 of life. Thus, hemipituitary glands were obtained from animals given two injections of OB, an injection of pregnant mare serum gonadotrophin (PMSG) or a unilateral brain lesion placed in the basal hypothalamus. Pituitary tissue was stimulated as before with a pulse of GnRH. Two injections of OB enhanced the sensitivity to stimulation. Conversely, both PMSG and lesion treatment severely reduced the sensitivity to GnRH, although PMSG-treated and lesioned animals have been used as models for the study of ovulation.


Trypanosoma cruzi: modification of macrophage function during infection. Infection of mice with Trypanosoma cruzi and subsequent intraperitoneal challenge with heat-killed trypanosomes elicits peritoneal macrophages which display in vitro microbicidal activity against trypomastigotes of T. cruzi. These cells also display other activated properties including rapid spreading, intense membrane activity, secretion of high levels of plasminogen activator, and ingestion mediated by the C3 receptor. An intravenous infection with BCG, followed by an intraperitoneal challenge with mycobacterial antigens brings about macrophages with similar properties. These criteria of macrophage activation were compared in normal and BCG- or T. cruzi-immune mice, with or without an intraperitoneal challenge with specific or unrelated antigens. Trypanocidal activity is displayed by both BCG- and T. cruzi-immune macrophages after intraperitoneal challenge with either antigen. Resident-immune macrophages from both T. cruzi- and BCG-infected mice show a trypanostatic, rather than trypanocidal activity. Macrophages from noninfected mice, challenged with the same antigens, show neither trypanostatic nor trypanocidal activity. Increased secretion of plasminogen activator shows a definite immunological specificity. Challenge with the specific antigen induces the appearance of macrophages secreting high levels of plasminogen activator, while unrelated antigens induce much smaller levels. Noninfected mice challenged with the same antigens do not display any enchancement in secretion. In contrast, increased spreading and phagocytosis mediated by the complement receptor are also displayed by cells from noninfected mice challenged with any of the agents tested.


Trypanosoma cruzi: the immunological induction of macrophage plasminogen activator requires thymus-derived lymphocytes. In this article we describe methods in which unstimulated mouse peritoneal macrophages were induced to secrete high livels of plasminogen activator under in vitro conditions. The exposure of sensitized peritoneal or spleen cell populations from Trypanosoma cruzi-infected animals to either viable or heat-killed trypanosomes lead to the release of an inducing factor(s). Maximal levels of plasminogen activator secretion are achieved by the incubation of such factors (s) with unstimulated macrophages for 48 h. A significant increase in enzyme secretion was already observed after a 24 h incubation. The production of the inducing factor(s) by sensitized cells was immunologically specific and unrelated antigens did not stimulate the production of the factor(s) by sensitized peritoneal or spleen cell populations. The inducing factor(s) was produced by nylon-wool-fractionated spleen and peritoneal cells which had been depleted of marcrophages. Pretreatment of sensitized spleen cells with anti-theta serum and C abolished the production of the activating factor(s). The active supernatant fluids were able to induce secretion of macrophage plasminogen activator across H-2 barriers. Attempts to induce trypanocidal activity in unstimulated macrophages have not been successful.


Evidence for differences in erythrocyte surface receptors for the malarial parasites, Plasmodium falciparum and Plasmodium knowlesi. Human erythrocytes lacking various blood group determinants were susceptible to invasion by Plasmodium falciparum including Duffy-negative erythrocytes that are refractory to invasion by Plasmodium knowlesi. Erythrocytes treated with trypsin or neuraminidase had reduced susceptibility of P. falciparum and normal susceptibility to P. knowlesi. Chymotrypsin treatment (0.1 mg/ml) blocked invasion only by P. knowlesi. The differential effect of enzymatic cleavage of determinats from the erythrocyte surface on invasion by these parasites suggests that P. falciparum and P. knowlesi interact with different determinants on the erythrocyte surface.


Streptococcal-induced cell-mediated-immune destruction of cardiac myofibers in vitro. We have demonstrated that T lymphocytes from the spleens of adult guinea pigs sensitized to group A streptococcal antigens are cytotoxic for cultured fetal guinea pig heart cells. Lymphocyte cytotoxicity, measured by 51Cr release from target cells, was stimulated by sensitization in vivo with group A whole cells, cell walls, and purified protoplast membranes emulsified with complete Freund's adjuvant (CFA). Sensitization with group C streptococcal antigens in CFA or CFA alone produced lymphocytes with little or no specific cytotoxic activity. Target cells of cultured fetal skeletal muscle, liver, or skin were relatively refractory to effector cell cytotoxicity. The presence of antigenic determinants on the membranes of cultured myofibers, cross-reacting with group A streptococcal cellular antigens, was confirmed by immunofluorescence. These data are discussed in terms of a model for poststreptococcal rheumatic myocarditis in which cell-mediated autoimmune mechanisms may participate.


Antigenic modulation of Friend virus erythroleukemic cells in vitro by serum from mice with dormant erythroleukemia. Friend leukemia virus (FLV) erythroleukemic cells cultured in medium containing FLV-immune serum from dormant FLV-infected mice undergo modulation of FLV cell surface antigens. Modulation was determined by an increased resistance to FLV antibody-mediated complement-dependent lysis and was associated temporally with the capping of FLV-immune complexes at the cell surface. Modulated cells regained their susceptibility to FLV antibody-mediated complement-dependent lysis when transferred to medium containing normal mouse serum. After 48 h of culture in FLV-immune serum, 26% of the FLV erythroleukemic cells were devoid of FLV cell surface antigens as demonstrated by immunofluoresence. Antigenic modulation occurred to a greater extent in cells maintained in logarithmic growth than in cells in GO or resting phase. FLV-antigenic modulation is discussed as a possible mechanism by which antibody induces and maintains FLV-transformed cells in a dormant state.


Pathogenesis of Shigella diarrhea. VII. Evidence for a cell membrane toxin receptor involving beta1 leads to 4-linked N-acetyl-D-glucosamine oligomers. The binding of ShigeUa dysenteriae 1 cytotoxin to HeLa cells in culture and to isolated rat liver cell membranes was studied by means of an indirect consumption assay of toxicity from the medium, or by determination of cytotoxicity to the HeLa cell monolayer. Both liver cell membranes and HeLa cells removed toxicity from the medium during incubation, in contrast to WI-38 and Y-1 mouse adrenal tumor cells, both of which neither bound nor were affected by the toxin. Uptake of toxin was directly related to concentration of membranes added, time,and temperature, and indirectly related to the ionic strength of the buffer used. The chemical nature of the membrane receptor was characterized by using three principal approaches: (a) enzymatic sensitivity; (b) competitive inhibition and (c) receptor blockade studies. The receptor was destroyed by proteolytic enzymes, phospholipases (which markedly altered the gross appearance of the membrane preparation) and by lysozyme, but not by a variety of other enzymes. Of 28 carbohydrate and glycoprotein haptens studied, including cholera toxin and ganglioside, only the chitin oligosaccharide lysozyme substrates, per N-acetylated chitotriose, chitotetraose, and chitopentaose were effective competitive inhibitors. Greatest inhibition was found with the trimer, N, N', N" triacetyl chitotriose. Of three lectins studied as possible receptor blockers, including phytohemagglutinin, concanavalin A, and wheat germ agglutinin, only the latter, which is known to possess specific binding affinity for N, N', N" triacetyl chitotriose, was able to block toxin uptake. Evidence from all three approaches indicate, therefore, existence of a glycoprotein toxin receptor on mammalian cells, with involvement of oligomeric beta1{arrow}4-1inked N-acetyl glucosamine in the receptor. This receptor is clearly distinct from the G(M1) ganglioside thought to be involved in the binding of cholera toxin to the cell membrane of a variety of cell types susceptible to its action.


C6: synthesis by the liver in vivo.

C6: synthesis by the liver in vivo. The allotype of the sixth component of complement was determined in a patient before and after liver transplantation. The C6 phenotype changed from A before transplantation to B (the donor phenotype) within 10 days of the transplant and remained wholly of the donor phenotype at 17 wk. This demonstrates that the liver is the exclusive or predominant site of C6 synthesis in vivo in man.


Group A streptococcal antigens cross-reactive with myocardium. Purification of heart-reactive antibody and isolation and characterization of the streptococcal antigen. Heart-reactive antibody (HRA) appears in the sera of experimental animals inoculated with group A streptococci as well as patients with acute rheumatic fever. Adsorption of either serum with group A streptococcal membranes will remove the HRA. Blocking experiments between these two types of HRAs have demonstrated that the antibodies are directed towards different antigenic determinants on either the same or different molecules. To isolate and purify the antigen from the group A streptococcus cross-reactive with sarcolemmal sheaths of cardiac myofibers, it became necessary to purify the HRA from rheumatic fever patients' sera. Isolated gamma globulin containing all of the HRA was adsorbed onto human sarcolemmal sheaths. The specific HRA was released by using potassium iodide. Over 99 percent of the purified HRA was shown to bind the sarcolemmal sheath whereas less than 1 percent of the antibody would bind nonspecifically to other material. Preparations of group A streptococcal membrane will bind HRA purified from the sera of acute rheumatic patients at levels of 97 percent or greater. The cross-reactive antigen solubilized by nonionic detergent was purified 120-fold by column chromatography. On sodium dodecyl sulfate polyacrylamide electrophoresis, the antigen was demonstrated to be composed of four polypeptides with mol wt of 32,000, 28,000, 26,000, and 22,000 daltons, respectively. Only proteolytic enzymes could destroy the antigenic determinant whereas glycosidases and lipases had no effect. The purified antigen blocked the binding of purified HRA to normal human heart sections.


Biochemical genetics of the alpha-keto acid dehydrogenase complexes of Escherichia coli K12: isolation and biochemical properties of deletion mutants. Mutants of Escherichia coli K12 with deletions in the nadC-lpd region of the chromosome were obtained for use in studies on the expression of the ace (pyruvate dehydrogenase complex, specific components) and lpd (lipomide dehydrogenase) genes. These were isolated by selecting spontaneous aroP mutants (lacking the general aromatic amino-acid permease and thus resistant to inhibitory aromatic amino-acid analogues) and screening for auxotrophy due to deletions extending into neighbouring genes. From 2892 isolates tested, the AroP- phenotypes of 2322 were confirmed and, of these, 28 stable and independently-derived auxotrophos were designated as deletion mutants. Six nutritionally-distinct categories were recognized: Nad- (8 strains); Nad-Ace-(7): Nad-'Ace-' (3); Ace- (8); 'Ace-' (I); Lpd-(I). The Ace- phenotypes of four isolates designated 'Ace-' were leaky and enzymological studies confirmed that they had less than 7% of parental pyruvate dehydrogenase complex activity. Enzymological studies showed that the 15 Ace- or Nad-Ace- strains all lacked the pyruvate dehydrogenase complex and pyruvate dehydrogenase (EIp) activities and only three retained detectable dihydrolipoamide acetyltransferase (E2p). The one Lpd- strain lacked pyruvate dehydrogenase, dihydrolipoamide acetyltransferase and lipoamide dehydrogenase (E3) activities as well as the activities of the pyruvate and alpha-ketoglutarate dehydrogenase complexes. The results confirmed the gene order nadC-aroP-aceE-aceF-lpd and indicated that no other essential functions are determined by genes within the nadC-lpd region. Resistance to lactate during growth of pps mutants on acetate was directly related to the specific activity of the pyruvate dehydrogenase complex. None of the deletions promoted the high degree of resistance characteristically associated with constitutive expression of the dehydrogenase complex. Six pps mutants having Ace+ or 'Ace-' phenotypes were more sensitive than the parental strains and expression of their ace operons appeared to be affected; most sensitive were the Ace- strains which lacked pyruvate dehydrogenase complex and phosphoenolpyruvate synthetase activities. The lipoamide dehydrogenase activities of the deletion strains (Lpd+) ranged between 30% and 100% of parental levels indicating that expression of their ace operons appeared to be affected; most sensitive were the Ace- strains which lacked pyruvate dehydrogenase complex and phosphoenolpyruvate synthetase activities. The lipoamide dehydrogenase activities of the deletion strains (Lpd+) ranged between 30% and 100% of parental levels indicating that expression of the lpd gene may be affected by the ace operon but can be independent.


Induction of cell division in a temperature-sensitive division mutant of Escherichia coli by inhibition of protein synthesis. Synchronous cells of the thermosensitive division-defective Escherichia coli strain MACI (divA) divided at the restrictive temperature (42 degrees C) if they were allowed to grow at 42 degrees C for a certain period before protein synthesis was inhibited by adding chloramphenicol (CAP) or rifampicin. The completion of chromosome replication was not required for such divA-independent division. Synchronous cells of strain MACI divided in the presence of an inhibitor of DNA synthesis, nalidixic acid, if they were shifted to 42 degrees C and CAP or rifampicin was added after some time; cells of the parent strain MC6 (div A+) treated in the same way did not divide. These data suggest that coupling of cell division to DNA synthesis depends on the divA function. The ability to divide at 42 degrees C, whether or not chromosome termination was allowed, was directly proportional to the mean cell volume of cultures at the time of CAP addition, suggesting that cells have to be a certain size to divide under these conditions. The period of growth required for CAP-induced division had to be at the restrictive temperature; when cells were grown at 30 degrees C, in the presence of nalidixic acid to prevent normal division, they did not divide on subsequent transfer to 42 degrees C followed, after a period, by protein synthesis inhibition. A model is proposed in which the role of divA as a septation initiator gene is to differentiate surface growth sites by converting a primary unregulated structure, with the capacity to make both peripheral wall and septum, to a secondary structure committed to septum formation.


Preliminary characterization of cell-free K99 antigen isolated from Escherichia coli B41. The K99 antigen of Escherichia coli B41 was isolated by isoelectric precipitation from heated bacterial suspensions. Chromatography and immunoabsorption experiments suggested that the mannose-resistant haemagglutinating activity of partially purified preparations of antigen was K99. The antigen was partially susceptible to bacterial proteases and was inactivated by periodate oxidation. Haemagglutination inhibition experiments with sugars and absorption of K99 with antisera to human blood groups A and B substances suggested that K99 contains a terminal alpha-linked N-acetylgalactosamine moiety, which is involved in the haemagglutination reaction, and an adjacent terminal alpha-linked galactose moiety, which plays no part in the reaction.


The influence of growth substrate and capacity for oxidative phosphorylation on respiratory oscillations in synchronous cultures of Escherichia coli K12. Fluctuations in cell volume during exponential growth of Escherichia coli K12 changed the effectiveness of the continuous-flow centrifugation method for preparing synchronous cultures. Rates of oxygen uptake in synchronous cultures were measured using an electrode system open to the atmosphere. In synchronous cultures of both the parental strain and an adenosine triphosphatase-deficient mutant, which was incapable of oxidative phosphorylation, respiration rates doubled during the cell cycle but oscillated with a periodicity of approximately half a cycle. Synchronous cultures of the parental strain growing on glycerol and Casamino acids showed a stepwise pattern of oxygen consumption. Continuous flow centrifugation did not markedly affect the increases in the numbers and respiration rates of cells in syndhronous cultures. Respiratory oscillations also occurred on inoculation of a late-stationary phase culture into fresh medium, although synchronous division was not observed. The possible mechanisms underlying respiratory fluctuations under different growth conditions are discussed.


Cytochemical differentiation of the axon membrane in A- and C-fibres. Guinea pig and rat sciatic nerves were fixed with cacodylate-buffered aldehydes and OsO4, and were stained with ferric ion and ferrocyanide. Cytoplasmic surfaces of the non-myelinated nodal axon membrane of A-fibres display distinct electron-dense aggregates of stain. These aggregates were not observed in association with the paranodal or internodal axolemma. The membranes of C-fibres exhibit no staining under these conditions. Thus, the nodal axolemma of normal myelinated fibres is structurally distinct from both the myelinated internodal membrane, and from the axolemma of C-fibres. The ferric ion-ferrocyanide technique may provide a method for marking axonal membrane with normal nodal properties.


Iron-acrylic compound for steriotaxic aneurysm thrombosis. A new iron-acrylic compound has been developed for stereotaxic thrombosis of intracranial aneurysms. The compound polymerizes rapidly, does not fragment, and is nontoxic. It has been used in a series of experimental animals and in initial clinical cases with good results. The use of this material simplifies and increases the safety of stereotaxic aneurysm treatment.


Pain reduction by electrical brain stimulation in man. Part 1: Acute administration in periaqueductal and periventricular sites. Acute studies performed in five patients indicate that electrical stimulation of the brain could be a powerful tool for the reduction or control of intractable pain. While chronic or spontaneous pain could be relieved by stimulation of the periaqueductal gray matter, the accompanying side effects render it impossible to stimulate this site regularly. On the other hand, stimulation of medial thalamic sites, particularly medial to the nucleus parafascicularis, yielded good relief of chronic pain at parameters which did not cause many undesirable side effects. The same parameters also produced inhibition of acute pain in two of the five patients.


Effects of positive end-expiratory pressure on intracranial pressure and compliance in brain-injured patients. Hypoxic pulmonary disorders and head injuries associated with increased intracranial pressure (ICP) frequently co-exist. Positive end-expiratory pressure (PEEP) improves hypoxemia but has been reported to impede cerebral venous return, potentially causing a further increase in ICP. This study examined the effects of PEEP on ICP at different levels of brain compliance. continuous ICP recordings were obtained after insertion of Scott cannulas to the lateral ventricles of seven comatose patients. Brain compliance was assessed by calculation of the pressure volume index. Patients were maintained in a 30 degrees head-up position. Maintenance of PEEP to levels of 40 cm H2O pressure for as long as 18 hours did not increase ICP in patients with either normal or low intracranial compliance, and did not increase ICP in the absence of pulmonary disease. Central venous pressure and pulmonary artery wedge pressure increased proportionately as PEEP was increased. No consistent changes were found in blood pressure recordings, nor were there any reductions in cardiac output found during the studies. Abrupt discontinuation of PEEP did not result in increased ICP except for a transient rise on two occasions when respiratory secretions became copious and the patients were inadequately ventilated. Improved oxygenation in two patients as a result of PEEP was concomitant with improved intracranial compliance and neurological status. In patients with brain injuries, PEEP improves arterial oxygenation without increasing ICP as previously supposed. Consequently, PEEP is a valuable form of therapy for the comatose patient with pulmonary disorders such as pneumonia or pulmonary edema.


Further characterization of rennin action on kappa-casein using carboxymethylcellulose: effects of various additives on the enzymatic hydrolysis of kappa-casein. The effects of various additives on the reaction of rennin with kappa-casein were investigated by using carboxymethylcellulose. Both urea and sodium 1-anilino-8-naphthalenesulfonate effectively inhibited rennin action at concentrations larger than 2 M and 2 mM, respectively. These reagents, however, activated the enzyme action at the lower concentrations. Both alpha S-and beta-caseins had some ranges of concentrations in which the rennin reaction was activated. Calcium chloride had an inhibitory effect on the rennin action. Neither mercaptoethanol nor KCl had any appreciable effect on the enzymatic hydrolysis of kappa-casein. These results are analyzed in terms of the association and dissociation of kappa-casein due to the presence of these additives in the reaction solutions.


Toxic neuropathy--an overview.

Toxic neuropathy--an overview. The involvement of the nervous system in systemic intoxication offers diagnostic and investigative challenges to the occupational health team. Often early evidence of neurotoxicity is vague, of a highly subjective nature and may be complicated by the presence of neuropathies due to other causes such as diabetes mellitus or exposure to multiple neurotoxins such as lead and solvents. The variety of agents producing neurotoxicity is wide, including heavy metals, organophosphates, acrylamide and its analogs, industrial solvents, therapeutic agents and plant alkaloids. The identification of neurotoxic substances requires well controlled epidemiologic studies of humans, including electrodiagnostic testing and well planned experimental animal studies. The latter include consideration of choice of animal species, routes of administration, duration of exposure, clinical evaluation, electrodiagnostic testing, morphologic description of the damage produced and its distribution, and the recognition of the potential for intercurrent naturally occurring neuropathologic processes. The aim of this discussion is to give an overview of these and other factors involved in the assessment of a neurotoxic potential.


Preprosthetic augmentation grafting--autogenous bone. Alveolar bone graft augmentation has returned successful prosthetic denture function to patients with deficient alveolar ridges who were beyond the scope of treatment by preprosthetic soft tissue surgical procedures alone. The results of a 30.7-month mean postoperative follow-up of 15 patients with panoramic radiographs were recorded. Emphasis was placed on evaluation by functional improvement rather than on radiographic interpretation. Ideal results were not obtained in all instances; however, in the majority of cases, the patient benefitted by improved prosthetic function.


The use of a modified skin grafting technique for alveolar sulcus extension. A modified technique for alveolar sulcus extension was developed from the classic procedure of extending the oral sulcus with epithelium. The ease of surgical manipulation sharply reduces postoperative morbidity and offers highly satisfactory functional results.


Current concepts of the adult respiratory distress syndrome. The adult respiratory distress syndrome (ARDS) is a sequel to pulmonary injury that may be direct, closed chest trauma or indirect, through air or vascular passages, aspiration, or fat embolization. An understanding of this syndrome is essential for the oral surgeon who not only manages severe maxillofacial injuries but is also a member of a trauma team that manages multisystem injuries. Emphasis on pathophysiologic pathways resulting in ARDS is presented with a discussion on oxygenation and ventilation abnormalities. Application of these guidelines will assist the oral surgeon in understanding the management of patients with this acute progressive syndrome.


Characteristics of mononuclear cell populations in chronically inflamed synovial membranes. Mononuclear cells infiltrating synovial membranes in chronic synovitis were characterised both in situ and in cell suspensions by surface markers and histochemical techniques. T-lymphocytes were the predominant infiltrating cell in rheumatoid arthritis as well as in other forms of chronic arthritis, including ankylosing spondylitis and arthritis associated with Crohn's disease. B-lymphocytes were found exclusively in rheumatoid synovial membranes. These cells were demonstrable both in true germinal centres and, focally and diffusely, in nodular mononuclear infiltrates lacking the histochemical characteristics of germinal centres. The synovial lining cells, unlike mononuclear phagocytes, had no demonstrable receptors for C3 and Fc.


The locomotory behaviour of the multinucleate giant cells of foreign body reactions. Cinemicrographic observations have shown that multinucleate giant cells are motile but the degree of translocational movement decreases with increasing size. Generally these polykaryocytes are slower than macrophages exhibiting a speed translocation of 0-3-1-0 micron/min for small polykaryocytes (2-4 nuclei) to 0-04 micron/min for large polykaryocytes (greater than 15 nuclei). Such behaviour would tend to maintain these cells near the site of their formation. The direction of translocation of small syncytia, correlates with the direction of the single leading lamellipodium. Slightly larger cells (5-15 nuclei) show loss of polarity with multiple lamellipodia, none of which is dominant, while the very large syncytia possess a single, large, circumferential lamellipodium whose movement leads to little displacement. These latter polykarya also display reduced contact inhibition which may favour fusion phenomena. Glycolysis appears to be the main source of energy for cellular movement which is also dependent on physiological calcium concentration and an intact microtubular-microfilamentous system. Apposition to the substratum is not complete, occurring in some instances by means of flaps and microvilli which may also be involved in cellular locomotion. Loss of glycocalyceal material probably occurs during locomotion.


Amalgam tattoos: light and electron microscopy and electron-probe micro-analysis. A common tattoo occurring in the mouth is caused by the insertion of amalgam filling material into the soft tissues. Fifteen amalgam tattoos were examined. Amalgam or its derivatives were found within macrophages, fibroblasts and multinuclear giant cells. Fine particles were found associated with: the basement-membranes of mucosal epithelium, of striped muscle fibres, and of muscle cells of blood vessels; collagen; elastic tissue; and the connective tissue of nerves. Amalgam consists mainly of mercury, silver and tin. The analytical results suggest that corrosion occurs and that mercury and some tin are lost from the tattoos, leaving silver and tin in macrophages and giant cells, and silver in fibroblasts and in the above-mentioned extracellular sites.


Jejunal mucosal immunoglobulins and complement in untreated coeliac disease. Jejunal immunofluorescence studies have shown that there is a significantly increased incidence of extracellular IgA and complement in the basement membrane zone and lamina propria of untreated adult coeliac patients compared to coeliac patients on a gluten-free diet. IgG was also demonstrated with complement, particularly in the untreated patients. These findings, taken together with those of other studies, suggest that a local antibody-antigen reaction involving IgA and complement may be responsible for the ongoing mucosal damage in untreated coeliac disease as well as for the acute damage following gluten challenge of the treated patient. Furthermore, consequent upon this damage there may be a secondary IgG antibody response, possibly reticulin antibody, contributing to the mucosal damage.


Ig surface receptors and erythrophagocytic activity of histiocytosis X cells in vitro. Membrane receptors for Ig and IgG and receptors for immune complexes (C3) have been identified on the surface of histiocytosis X cells by direct immunofluorescence and by rosette formation. Histiocytosis X cells also avidly phagocytose human RBC and latex particles. The cells studied originated from three eosinophilic granulomas of bone and were maintained in culture for 12 days. These immunological characteristics suggest that histiocytosis X cells represent a neoplastic variant of a cellular subpopulation (Langerhans lineage) which probably belongs to the mononuclear phagocyte system.


Abnormalities of the local immune system in intestinal neoplasia: a morphological study. In colonic adenocarcinomata abnormalities have been noted in all morphological aspects of the local immune system of the bowel; thus, in such tumours there is a defective production of secretory IgA, a diminished content of secretory component (SC) and a marked decrease in intraepithelial lymphocytes. By contrast, no such abnormalities were noted in the non-neoplastic areas of the bowel, either immediately adjacent to, or some distance from, the carcinoma. These findings suggest that there is not usually any generalised morphological abnormality of the local immune systems in intestinal neoplasia and that the immunological abnormalities noted within the tumour are a result of the changed nature of the epithelium, thus altering, or interfering with, the normal interactions between epithelial and lymphoid tissue in the gut.