The amino acid sequence of the ribosomal protein S8 of Escherichia coli. The primary structure of protein S8 from the 30S subunit of Escherichia coli ribosomes has been determined by sequencing the peptides derived from tryptic, chymotryptic, thermolytic and staphylococcal protease digestion of the protein. Protein S8 has 129 amino acid residues which result in a molecular weight of 13996. The N-terminal part of the sequence up to position 68 is in complete agreement with the reported sequence data[1,2]. However, differences exist in the C-terminal half, where an additional hydrophobic tryptic peptide has been found.


Correlation of carcinoembryonic antigen in tissue sections with spread of mammary carcinoma. An immunoperoxidase technique for the histological demonstration of Carcinoembryonic Antigen (CEA) was applied to paraffin sections from 74 breast carcinomas and 43 benign breast lesions. Sixty-six per cent of the carcinomas and the only case of granular cell myoblastoma examined were CEA positive. Two examples of mammary dysplasia (7%) showed foci of CEA positive acini. All tumour tissue found in lymphatics and in metastases in lymph nodes was CEA positive, including two cases where the primary tumour was CEA negative, and all the metastases examined from CEA positive tumours. A significant relationship was found between CEA positivity of the primary tumour and the presence of lymph node metastases.


Using the Federal Register.

Using the Federal Register. The Federal Register provides official notice of the issuance and contents of agency regulations and proposed rule changes and gives citizens an opportunity for input into their development. Knowing how to use this publication should be part of any administrator's repertory of skills.


Stage classification of nasopharyngeal carcinoma: a review. Various stage classifications of NPC have been described and discussed. Treatment results have been reported for the various stages for only four of these classifications; the correlation between stage and treatment results was best in the classification of the author.


The Cri du Chat syndrome: epidemiology, cytogenetics, and clinical features. Data for 331 cri du chat cases, including 34 Danish probands, are reviewed. The incidence nad the prevalence among the mentally retarded population amounted to 1/45,000 and 1.5/1000, respectively. No striking association with prenatal events, parental ages, or birth order could be demonstrated. There was  a significant excess of females. Parental translocations were present in slightly more than 10% of the families, while more rare cytogenetic aberrations (mosaicism, rings, and de novo translocations) accounted for less than 10% of all cases. The phenotypically relevant segment has been narrowed down to the midportion of the 5p15 band. Clinical, radiologic, and dermatoglyphic features are summarized and discussed, with special attention to the abnormal cry, which persists in many older probands, and to developmental abnormalities. No obvious correlation could be detected between clinical features and the localization of the deletion. No marker locus has yet been assigned to the short arm of chromosome 5. Treatment and prevention are briefly discussed.


Production of erythropoietin in vitro: a review. The in vitro production of the important regulator of erythropoiesis, erythropoietin (Epo), is reviewed. It is concluded that it is possible to produce almost routinely small quantities of Epo in tissue culture. Although such procedures offer the potential to provide large quantities of the hormone for clinical use, the optimum culture conditions and mechanisms for triggering Epo production have yet to be resolved.


[Comparative susceptibility testing to sulfamethoxazole and trimethoprim by agar diffusion test on different media (author's transl)]. 14 lots of 6 different commercial media for susceptibility testing were compared by agar diffusion test with 161 bacterial strains. The result "resistant" to sulfamethoxazole, trimethoprim, or co-trimoxazole varied in wide limits between the media. The lot employed by the "Work-Group on Resistance" of the Paul Ehrlich Society is of limited value for testing of the investigated drugs.


[Inoculum effects and bacterial resistance (author's transl)]. Massive bacterial populations are frequently involved in infection. Such dense cultures may appear 'resistant' to therapeutically achievable concentrations of some antibacterial agents in vitro, but the implications of this for treatment are not always clear. A better understanding of the mechanisms responsible for inoculum effects may help in assessing their clinical significance and some possible approaches to this goal are described.


[Susceptibility testing of anaerobes (author's transl)]. The methodology of susceptibility testing of anaerobes has not yet been standardized and many individual test methods are in use. Among the factors which may influence the outcome of susceptibility tests are test organisms, antimicrobials and methods of susceptibility testing. Comparative studies have shown that the density of the inoculum largely influences the MICs of sulphonamides and certain other antimicrobials. In testing susceptibility of anaerobes to doxycycline, the MICs determined by agar dilution tests were 2--4--8 times the MICs obtained by broth dilution technique. However, with certain cephalosporins and beta-lactamase-producing strains of Bacteroides fragilis and other Bacteroides species the MICs obtained by the two methods differed by four to five dilution steps. Judging by the broth dilution test, the majority of the beta-lactamase-producing Bacteroides strains would be classified as resistant, whereas with beta-lactamase-negative strains of Sphaerophorus varius the broth dilution test gave much lower MICs than the agar dilution technique. These findings pose the rather embarrassing problem as to whether strains are to be regarded as resistant or sensitive. As is demonstrated by lack of stochastic linearity and/or low correlation coefficients, the results of standardized anaerobic agar diffusion tests are for the most part not correlated to MICs determined by either method of quantitative susceptibility testing.


[Microcalorimetric Investigations, regarding the antibacterial efficiency of chemotherapeutics (author's transl)]. An investigation method is described by which microcalorimetry and continuous density measurement are used to show the antibacterial efficiency of chemotherapeutics. Oleandomycin, erythromycin, both macrolide antibiotics, and doxycycline, a tetracycline, blockers of the protein synthesis, have the same effect on the microcalorimetric curve of a staphylococcus strain. Among the aminoglycosides tobramycin and gentamicin influenced the microcalorimetric activity of a growing E. coli strain in the same way, spectinomycin produced different curves. Penicillins interfere with the synthesis of the cell membrane by blocking the enzyme transpeptidase. Epicillin, penicillin, ampicillin, oxacillin, carbenicillin, and azlocillin influence the microcalorimetric activity of staphylococci in a typical way which is different from that of the blockers of the protein synthesis.


Internal fixation of fractures of the neck of the femur using von Bahr screws and allowing immediated weight bearing: a prospective clinical study. This article presents the results of von Bahr screw fixation in 103 patients, allowing immediate weight bearing. After a follow-up period of 1 year, a failure rate of 18% was recorded. Analysis of the series shows that the failures are due to inadequate reduction of the displacement during operation, especially when the head is in varus, and to bad positions of the screw. The high failure rate was not related to the initial displacement of the fracture.


Plasmid carriage and the serum sensitivity of enterobacteria. The carriage of a range of plasmids by rough, serum-sensitive laboratory strains of Escherichia coli made no difference to their reactivity in human serum as determined by two methods. Plasmid-carrying enterobacteria isolated from polluted river water gave a variety of responses to serum. Smooth E. coli river isolate C8 was killed by serum but only after a delay of 1 h, and curing of antibiotic resistance and colicin determinants from this strain led to a small but significant increase in serum sensitivity. Plasmids from eight strains were transferred by conjugation to a cured derivative of C8 (C8(-)Nal(R)), and in six cases a significant increase in the serum resistance of the progeny was observed. Plasmid-mediated enhancement of resistance was particularly marked with plasmids R1 and NR1, and a round of replication mutant of NR1 conferred greater resistance than did the normal R factor. However, R1 and NR1 were unable to modify the serum response of a cured strain (P21(-)Nal(R)) derived from promptly serum-sensitive isolate P21. These findings suggest that lipopolysaccharide O-side chains, the cell surface components responsible for the delay in serum killing, are essential for the expression of plasmid factors that modify sensitivity to serum. Examination of K(A)(-) variants of two isolates indicated that the K(A) antigen has only a marginal effect on the serum response.


Protective effects of a supernatant factor from Salmonella typhimurium on Salmonella typhimurium infection of inbred mice. A supernatant factor prepared from 48-h cultures of Salmonella typhimurium has been used to immunize mice against subsequent challenge with normally lethal doses of S. typhimurium. The mouse strains used, C57BL and BALB/c, were sensitive to S. typhimurium with 50% lethal doses of less than 50 organisms. Two doses of supernatant factor, given intraperitoneally 20 days apart, protected mice against a subcutaneous challenge dose 10 days later of 100 50% lethal doses of S. typhimurium, resulting in 50 to 80% survival. The viable counts were reduced initially in organs of immunized mice compared with controls, and the multiplication of bacteria was delayed, although the final levels found in the organs would normally have been lethal. Protection obtained was specific for S. typhimurium in that no increased survival was shown after Salmonella enteritidis challenge of immunized mice. Although lipopolysaccharide was demonstrated in the supernatant factor, lipopolysaccharide alone did not protect challenged mice. Supernatant factor produced delayed-type hypersensitivity reactions in mice sensitized with nonlethal doses of Salmonella. The nature of the active factor, found to be partially protein, has yet to be elucidated.


Inhibition of the secretory activity of Escherichia coli heat-stable enterotoxin by indomethacin. The effect of indomethacin on the net intestinal accumulation of fluid induced by Escherichia coli heat-stable (ST) enterotoxin in the infant mouse model was examined. Indomethacin, when administered with ST enterotoxin, caused a striking decrease in net intestinal fluid accumulation. This inhibition of ST activity was dose dependent with various concentrations of indomethacin (P less than 0.01). A significant inhibition of toxicity was also observed when indomethacin was given before (P less than 0.01) or after (P less than 0.02) ST enterotoxin challenge. No significant differences in fluid accumulation were observed between control mice treated with buffer alone and those challenged with only indomethacin. These data indicate that indomethacin markedly decreases the net intestinal fluid accumulation induced by E. coli ST enterotoxin. Further studies on the potential use of indomethacin in both the prophylaxis and the therapy of diarrheal diseases appear warranted.


DNA release as a direct measure of microbial killing by phagocytes. A new assay for the precise measurement of microbial killing by leukocytes is presented. The method assumes that release of radioactively labeled DNA from the microbe is direct evidence of cell death. Human peripheral blood leukocytes incubated with [14C]thymidine-labeled Salmonella typhimurium released 32 to 59% of the radioactivity after 4 h and 63 to 75% after 18 h. Inactivated leukocytes released less than 5% of the radioactivity. None of the released radioactivity is retained within the leukocyte, and 60% remains precipitable with trichloroacetic acid. Leukocytes released substantial radioactivity from labeled Escherichia coli but only a slight amount from staphylococci. Mouse peritoneal macrophages were also shown to release radioactivity from Salmonella. The DNA release assay avoids the errors inherent in prior killing methods which measure viability by growth inhibition. It is rapid, reproducible, and highly specific.


Detection of antibodies to Mycoplasma pulmonis by an enzyme-linked immunosorbent assay. The enzyme-linked immunosorbent assay, which entails the use of antigen-coated tubes and enzyme-labeled anti-immunoglobulins, was applied for the detection of antibodies against Mycoplasma pulmonis in mice. A lysate of M. pulmonis was used as the antigen, and anti-mycoplasmal antibodies of the different immunoglobulin classes were detected by class-specific anti-immunoglobulin labeled with alkaline phosphatase. The optimal conditions for the test were determined, the specificity was evaluated, and the assay was compared with other procedures for detection of mycoplasmal infection. The enzyme-linked immunosorbent assay was found to be a specific, highly sensitive, and reliable procedure for detecting anti-mycoplasmal antibodies in mice.


Genetic separation of serum opacity factor from M protein of group A streptococci. Mutagenized cultures of three strains of group A streptococci which were M positive and produced the extracellular serum opacity factor (OF) were screened for mutants which failed to exhibit OF activity on serum agar plates. All cell fractions from three mutants studied in detail, including culture supernatants, protoplast membranes, and cytoplasmic fractions, were completely devoid of activity. Two OF- isolates also lacked OF antigen, whereas the third continued to produce cross-reacting antigen. All three mutants were resistant to phagocytosis and yielded acid-extractable M antigen, indicating that the M protein of each strain was unaltered by the mutations. The separation of the OF+ and M+ phenotypes by mutation establishes that genes which code for the M protein and OF are distinct; therefore, the activities and antigenic determinants of each must be associated with separate, distinct protein molecules.


Regional T- and B-cell responses in influenza-infected ferrets. Ferrets were infected with A/Port Chalmers/72 influenza virus and the T- and B-cell responses in the spleen, in lymph nodes draining the upper and lower respiratory tract, and in lung washings were examined in vitro. Lymphocyte responses were measured by using a hemolytic plaque assay for B cells and a proliferation assay for T cells. Virus and antibody levels were measured in respiratory tract washings, and antibody titers were measured in sera from infected animals. Individual B cells secreting specific antibody to A/Port Chalmers/72 virus were detected in regional lymph node and spleen preparations as early as 3 days and as late as 43 days after infection. T-cell assays showed an in vitro response of lymph node cells to A/Port Chalmers/73 virus from day 6 to day 43. Virus was isolated from the respiratory tract up to 7 days after infection. Serum hemagglutination-inhibiting antibody was first detectable on day 6, with maximum titers reached by day 10. These results demonstrated that antibody production and a cellular immune responses were detectable at regional sites at a time when virus was still present and before serum antibody was measured.


Radiolabeling of Treponema pallidum (Nichols virulent strain) in vitro with precursors for protein and RNA biosynthesis. We observed uptake of [U-14C]serine, U-14C-labeled amino acid hydrolysates, and [2-14C]uracil by virulent Treponema pallidum in vitro for at least 96 h. No uptake of [2-14C]thymine, [1-14C]pyruvate, [U-14C]pyruvate, and [2-14C]uridine was detected. Treponemal protein and RNA biosynthetic activity was identified by erythromycin inhibition of amino acid and uracil uptake. Radioactivity due to uptake of radiolabeled amino acids by residual testicular cells in the cultures remained at background levels regardless of the presence or absence of cycloheximide. Accumulation of the radiolabeled substrates by T. pallidum proceeded at a linear rate for 48 to 96 h during incubation in vitro. The longevity of substrate uptake using the system of incubation described will facilitate future studies on the metabolism of the microorganism to help determine essential growth factors and environmental conditions for multiplication of T. pallidum in vitro.


Mannose binding and epithelial cell adherence of Escherichia coli. The mannose-binding activity of several isolates of Escherichia coli was monitored by aggregometry with mannan-containing yeast cells. The velocity of yeast cell aggregation was found to correlate with the ability of the organisms to adhere to human epithelial cells. Mannose or its derivatives specifically inhibited or reversed epithelial cell adherence and yeast cell aggregation. Most of the adherent bacteria could be displaced within 30 min from the epithelial cells with methyl alpha-d-mannopyranoside, but not with other sugars tested. Cultures of E. coli were fractionated into nonadherent and adherent populations by adsorption with epithelial cells followed by elution of the adherent bacteria with methyl alpha-d-mannopyranoside. When the methyl alpha-d-mannopyranoside-displaced organisms were washed free of the sugar, they exhibited a high degree of mannose-binding activity and were heavily piliated. In contrast, the nonadherent fraction of organisms lacked detectable mannose-binding activity and were devoid of pili. Our results suggest that the binding activity of a mannose-specific lectin on the surface of E. coli can be quantitated directly on intact organisms, and the observed variations in the amount of mannose-binding activity among human isolates accounts for the variation in adherence of the organisms to mannose residues on epithelial cells.


Effect of L-form Streptococcus pyogenes and of lipoteichoic acid on human cells in tissue culture. These studies showed the destruction of growing primary and established human cell lines with a predilection for the group A streptococci by an L-form of Streptococcus pyogenes adapted to grow in isotonic media. Also, this L-form was detected by fluorescent antibody for longer periods of time than by viable count in infected but recovered tissue culture monolayers. Additional studies with human heart cells showed changes in their protein profile and fatty acid content (but not composition) after L-form infection. This report is the first to show that the morphological changes and death of human kidney cells by this viable L-form were mimicked by the structurally different lipoteichoic acids from this organism and its parental streptococcus. These lipoteichoic acids were also equally effective in preventing attachment of S. pyogenes to human cell monolayers, but their deacylation obviated these two activities. Finally, the attachment of the isotonic L-form, as well as the parental streptococcus, to growing human kidney cells suggested that a rigid cell wall is not a prerequisite for host attachment in vitro.


Activation of the alternative complement pathway by a streptococcal lipoteichoic acid. A glycerol lipoteichoic acid antigen from Streptococcus pyogenes 1-RP41 was found by rabbit erythrocyte hemolytic assay to activate the alternative complement pathway in human sera. Over a narrow concentration range of the teichoic acid, complement consumption was dose dependent, whereas at higher concentrations of the acid complement consumption could not be detected.


Comparison of the histamine hypersensitivity and the Limulus amoebocyte lysate tests for endotoxin activity. The histamine hypersensitivity test and the Limulus amoebocyte lysate test were compared for their effectiveness to quantitate endotoxin activity. The two tests compared favorably in all the trials, except with a sample of endotoxin from Brucella abortus that gave a positive Limulus amoebocyte lysate test at a concentration of 0.001 microgram, while failing to sensitize mice to histamine at a dose of 16 microgram per mouse. The Limulus amoebocyte lysate test was more sensitive than the histamine hypersensitivity test.


Antibody-independent interaction of the first component of complement with Gram-negative bacteria. The interaction of the first component of complement with two serum-sensitive strains of Escherichia coli and Klebsiella pneumoniae was studied. It could be demonstrated that highly purified C1, free of immunoglobulin G and immunoglobulin M, binds to E. coli or K. pneumoniae. C1 binding was also found with specifically absorbed human serum, after incubation of bacteria with normal serum in the presence of ethylenediaminetetraacetate or agammaglobulinemic serum; the number of C1 molecules taken up by the bacteria was not influenced, indicating that C1 binding was independent of naturally occurring antibodies. C1 bound to bacteria was still able to cleave C4, the natural substrate of C1. From these observations, it is concluded that C1 in an enzymatically active state can be bound directly to bacteria independently of antibody.


Delayed-type hypersensitivity responses in infected mice elicited by cytoplasmic fractions of Cryptococcus neoformans. Four subcellular fractions of Cryptococcus neoformans prepared by differential centrifugation of disrupted whole yeast and a 3-day culture filtrate were examined for their ability to elicit delayed-type hypersensitivity in sensitized animals. The methods used to detect sensitization were (i) the footpad swelling test and inhibition of peritoneal macrophage migration in mice and (ii) skin testing in guinea pigs. Two entities, the post-mitochondrial supernatant and the culture filtrate, showed considerable activity in the footpad test, with 26- and 30-microliter 24-h swellings, respectively, at 6 weeks after infection. With the latter there was interference from a strong antibody-mediated 4-h skin reaction. The post-mitochondrial supernatant produced strong delayed-type hypersensitivity in guinea pigs at a dose of 69 microgram, and there was no demonstrable cross-reactivity in animals sensitized with heterologous fungi. The footpad swelling in mice correlated well with the macrophage migration inhibition test, with 71% inhibition in mice infected subcutaneously with C. neoformans at 6 weeks. However, mice infected intravenously developed poorer cell-mediated immunity than the subcutaneously infected mice. The post-mitochondrial supernatant was found to contain detectable amounts of cryptococcal capsular polysaccharide.


Heat stability of Mycobacterium leprae immunogenicity. The protection provided to mice by vaccines administered intradermally was measured after footpad challenge with Mycobacterium leprae. The protection offered by M. leprae suspensions was not decreased when the vaccines were killed by 60 degrees C heat or at the higher temperatures tested, which included 215 degrees C (autoclave). Even highly purified suspensions retained their immunogenicity. In contrast, the vaccine protection provided by intradermal M. bovis (strain BCG) was markedly reduced when heated to 60 degrees C. The enlargement of the lymph nodes regional to the intradermal vaccines was measured and found generally to parallel the vaccine protection provided by M. leprae and by BCG.


Interaction of human serum and neutrophils with Escherichia coli strains: differences between strains isolated from urine of patients with pyelonephritis or asymptomatic bacteriuria. Strains of Escherichia coli isolated from the urine of children with pyelonephritis (n = 18) or asymptomatic bacteriuria (ABU) (n = 20) were serotyped and compared with respect to opsonic requirements, sensitivity to serum, amount of chemiluminescence produced by normal neutrophils during phagocytosis of the various strains, and amount of capsular antigen. Strains isolated from cases of ABU more often had the capacity to activate the alternative complement pathway than had strains causing pyelonephritis. The ABU strains were also better opsonized in fresh human serum and were more often serum sensitive than the pyelonephritic strains. There were individual variations in neutrophil chemiluminescence during phagocytosis of the various strains, but the two groups of strains did not differ significantly. The strains from pyelonephritis cases produced a greater amount of capsular antigens than the ABU strains. There was no correlation between presence of a particular O or K antigen and capacity to activate the alternative complement pathway. The observed differences between strains isolated from patients with ABU and those isolated from patients with pyelonephritis may be relevant for further studies on differences in bacterial virulence or host defense factors.


Alterations in the tRNA's of Escherichia coli recovered from lethally infected animals. Escherichia coli grown in chemically defined iron-deficient media or in fluids containing the iron-binding proteins transferrin, lactoferrin, or ovotransferrin have well-characterized alterations in the chromatographic properties of tRNA's containing the modified nucleoside 2-methylthio-N6-(delta2-isopentenyl)-adenosine. The present work shows that similar tRNA alterations occur in E. coli O111 recovered from the peritoneal cavities of lethally infected guinea pigs and rabbits. Adding iron to these in vivo-grown bacteria resulted in the rapid conversion of chromatographically abnormal tRNA's to the normal species. The work strongly suggests that host iron-binding proteins, present in mucosal and other secretions, can affect the metabolism of invading organisms. The idea that the tRNA alterations are connected with the adaptation of E. coli to growth under the iron restricted conditions imposed by iron-binding proteins in tissue fluids, and thus with bacterial pathogenicity, is therefore made particularly attractive.


Association between cross-reacting Escherichia coli K100 and disease caused by Haemophilus influenzae type b. An unexpected association between Haemophilus influenzae type b (HIB) disease and simultaneous intestinal colonization with cross-reacting Escherichia coli K100 strains is reported. E. coli K100 were found in 18 of 92 (19.5%) HIB patients, compared with 2.5% in healthy individuals of comparable age and in none of 21 patients with meningitis caused by other bacteria. There was no difference in disease outcome, serum antibodies, and circulating HIB antigen between the E. coli K100-positive and -negative HIB patients. Possible explanations and implications of this association are discussed.


Hyporeactivity factor produced after induction of immune interferon in mice sensitized with BCG. Sera from BCG-sensitized ICR and C57BL/10J mice challenged with BCG were examined. Two types of hyporeactive factor (HF), which produced refractoriness to induction of interferon (IF) by Newcastle disease virus in mouse embryo fibroblasts, were produced in addition to type II immune IF. One was destroyed at pH 2, and the other was not. The acid-labile HF, together with immune IF, appeared 2 h after challenge with BCG. Acid-labile HF and immune IF were not neutralized by antiserum against L-cell IF and may be one substance. On the other hand, acid-stable HF appeared later and was neutralized by antiserum against L-cell IF. Furthermore, acid-stable HF was more stable at 56 degrees C than were immune IF and pH-labile HF. Induction of IF by Newcastle disease virus was also reduced in mice 6 h after receiving one injection of BCG, irrespective of whether the animals were previously sensitized to BCG.


Role of bacterial products in periodontitis: humoral immune response to Eikenella corrodens. Eikenella corrodens can induce periodontitis-like disease in gnotobiotic rats. Some components of this bacterial cell elicit measurable humoral immune response during the development of the disease, but in this system endotoxin is not among the efficient immunogens. Because no humoral immune response could be seen to the endotoxin of Eikenella corrodens it is assumed that this endotoxin can act uncontrolled in monoinfected rats. Accordingly, the lack of protective humoral immune response to pathogenic components of Eikenella corrodens may be the major factor permitting the development of the disease described here. The possibility that both cell-mediated immunity and uncontrolled endotoxic action are parts of the pathomechanism of the disease is supported by our observations.


Adherence of an enteropathogenic strain of Escherichia coli to human intestinal mucosa is mediated by a colicinogenic conjugative plasmid. The capacity of a human enteropathogenic Escherichia coli (EPEC) strain serotype O26:K60:H11, to adhere to the mucosa of the human fetal small intestine was shown to be plasmid mediated. Adherence was transferred at a high frequency in a long-term conjugal mating experiment to E. coli K-12 and was lost by treatment of the EPEC strain with the curing agent ethidium bromide. Analysis of radioactively labeled DNA from lysates of the EPEC, transconjugant, and cured strains indicated that adherence was correlated with the presence of plasmid DNA species with an approximate average molecular weight of 56 X 10(6). Resistance to the antibiotics spectinomycin, streptomycin, sulfonamides, and tetracycline and production of colicin Ib were all transferred in long-term mating and lost upon curing coordinately with the property of adherence. In conjugal mating experiments of limited duration between E. coli K-12 strains, however, segregation of colicin production and mucosal adherence from multiple drug resistance was observed. Analysis of plasmid DNA of segregant transconjugant strains confirmed the presence in the 56 X 10(6)-dalton plasmid species of two previously unresolved components, pLG101 designating the ColIb plasmid which also carries the determinant for mucosal adherence and pLG102 representing the slightly smaller multiple drug resistance plasmid.


Differences in hydrophobic surface characteristics of porcine enteropathogenic Escherichia coli with or without K88 antigen as revealed by hydrophobic interaction chromatography. Porcine enteropathogenic Escherichia coli strains possessing or lacking K88 antigen were studied by using hydrophobic interaction chromatography on cross-linked agarose gels with alkyl or aryl substituents (amphiphilic gels) to determine whether or not they possessed surface-associated hydrophobic properties. Strains with K88ab or K88ac antigen adsorbed to phenyl and octyl Sepharose gels in the presence of 4 M sodium chloride. This property correlated with phenotypic expression of K88 antigen. Cells grown at 37 degrees C but not those grown at 18 degrees C possessed hydrophobic adsorptive characteristics in addition to the property of mannose-resistant hemagglutination of guinea pig erythrocytes. Adsorption of K88-positive strains to gels with hydrophobic ligands was independent of O group and enterotoxicity. Strains lacking K88 antigen did not adsorb to the hydrophobically substituted derivatives of Sepharose and lacked mannose-resistant hemagglutinating characteristics. Neither the presence of additional polysaccharide K antigens nor nonhemagglutinating pili conferred the property of hydrophobic interaction on the strains. K88-positive bacteria had a lower electrophoretic migration rate than did K88-negative bacteria of the same serotype in free-zone electrophoresis. K88-positive bacteria also adsorbed strongly to hydrophobic ligands in the presence of 1 M ammonium sulfate, whereas K88-negative strains did not. These observations provide evidence for the suspected role of hydrophobic interaction in the adhesive properties of certain enteropathogenic strains of E. coli. Moreover, hydrophobic interaction chromatography provides convenient and rapid alternative means of screening strains for a property potentially associated with adhesiveness.


Dynamics of Escherichia coli infection and meningitis in infant rats. Escherichia coli strains isolated from newborn infants were injected intraperitoneally into infant rats. Strains possessing the K1 capsular polysaccharide antigen were significantly more virulent than strains lacking this antigen. When 5-day-old animals were injected with 1.2 X 10(1) colony forming units of a K1 E. coli strain (serotype O18ac:K1:H7), about 80% had bacteria isolated from their blood. Forty-eight percent of bacteremic animals had positive cerebrospinal fluid cultures. The development of bacteremia with greater than 10(4) colony-forming units per ml of blood correlated with positive cultures of cerebrospinal fluid. Some animals, studied with serial blood cultures, were able to clear bacteria spontaneously from their blood, whereas others succumbed to infection within 48 h of challenge. The susceptibility of infant rats to E. coli infection was age dependent and appeared related to the K1 antigen.


Effects of injected iron and siderophores on infections in normal and immune mice. The fate of virulent and avirulent strains of Salmonella typhimurium in untreated and iron-injected mice and in transferrin-containing media demonstrated a direct relationship between bacterial virulence and the ability of bacteria to acquire transferrin-bound iron. Effects of injected iron on the development of infections with virulent and avirulent bacterial strains were determined in normal and immune mice by determinations of bacterial numbers in tissue homogenates and the mortality of infected animals. Results showed that infected and iron-injected mice died much more rapidly and frequently from overwhelming infections than infected and saline-injected mice. The infection-promoting effect of iron varied with the degree of bacterial virulence; the more virulent the bacteria, the more helpful was iron for the development of lethal infections. Siderophores promoted lethal infections in mice infected with virulent but not with avirulent bacteria. Experiments with vaccinated animals showed that iron exerted a deleterious effect on acquired immunity. Immune mice infected with virulent bacteria and injected with iron developed lethal infections as rapidly and nearly as frequently as similarly treated normal mice. Siderophores did not promote the development of lethal infections in immune mice. The effectiveness of iron, but not of siderophores, to promote bacterial infections in vaccinated mice revealed that acquired immunity is dependent upon the activity of an iron-neutralizable antibacterial system.


Growth of Mycobacterium lepraemurium in nonstimulated and stimulated mouse peritoneal-derived and bone marrrow-derived macrophages in vitro. Mycobacterium lepraemurium cells were found to multiply in normal mouse peritoneal-derived and bone marrow-derived macrophages in vitro. Whereas activated peritoneal-derived macrophages demonstrated marked bacteriostasis for M. lepraemurium, significant bactericidal activity was exhibited by activated bone marrow-derived macrophages. However, only a small proportion of the bacterial were killed by activated bone marrow-derived macrophages with subsequent and enhanced bacteria growth. It is suggested that a rapid turnover of monocytes in active lesions is required to control mycobacterial infections in vivo. These results would suggest that careful consideration be given to the choice of the host cell in studies involving obligate intracellular parasites.


Strain difference of delayed-type hypersensitivity to BCG and its genetic control in mice. Delayed-type hypersensitivity to BCG was remarkably different in two inbred strains of mice, SWM/Ms and C3H/He, when measured by the spleen index, the disappearance of peritoneal macrophage, or the footpad reaction. High responsiveness in the SWM/Ms strain appeared to be dominant over low responsiveness in the C3H/He strain. Results of the footpad reaction test if F1, F2, and backcross hybrids of these two strains of mice suggested that the delayed-type hypersensitivity was mainly controlled by a gene which was transmitted under Mendel's laws and was possibly non-H-2 linked. The spleen cells and their nylon wool nonadherent fraction from BCG-infected C3H/He mice were not reactive to purified protein derivative in vitro, whereas both the spleen cells and the nylon wool nonadherent fraction from BCG-infected SWM/Ms mice reacted well to purified protein derivative. Possible mechanisms of the different responses in the delayed-type hypersensitivity to BCG were discussed.


Nonspecific and specific immunological mitogenicity by group A streptococcal pyrogenic exotoxins. Group A streptococcal pyrogenic exotoxin (SPE) types A, B, and C induced lymphocyte proliferation both specifically and nonspecifically, and the responses showed characteristics associated with both types of stimulation. Guinea pig lymphocytes from animals presensitized to SPE A displayed immunologically specific proliferation in response to SPE A; control lymphocytes showed little activity in the presence of SPE A. Lymphocytes from guinea pigs not presensitized to SPE responded nonspecifically to SPE types B and C. Guinea pig lymphocytes from SPE A-presensitized animals showed enhanced proliferation over controls when treated with SPE B, suggesting that a degree of cross-reactivity between SPE types may exist, though they are serologically distinct. Mouse splenic lymphocytes exhibited low-level responsiveness to all SPE types, as would be expected for an antigen-specific proliferative response. Unlike mouse splenic lymphocytes, rabbit spleen cells and human cord blood, lymphocytes responded nonspecifically to all SPE types. Although rabbit spleen cells and human cord blood lymphocytes responded nonspecifically, the maximum response occurred at day 4 or 5, comparable to an antigen-specific system rather than a day 2 or 3 such as that with the nonspecific thymus-derived cell mitogen, concanavalin A.


Influence of oxygen tension, sulfhydryl compounds, and serum on the motility and virulence of Treponema pallidum (Nichols strain) in a cell-free system. The motility and virulence of Treponema pallidum (Nichols strain) were monitored during incubation in a modified tissue culture medium to study the effects of oxygen tension and medium composition on survival of the organism. A basal medium of Eagle minimal essential medium with 50% fresh, heat-inactivated normal rabbit serum was used inasmuch as better survival occurred with 50% normal rabbit serum than with lower concentrations. Addition of 0.5 to 2.0 mM dithiothreitol or 2.0 mM dithioerythritol to the basal medium led to significantly longer retention of T. pallidum viability in the presence of 3% oxygen than under aerobic or anaerobic conditions. The results of this investigation lend support to the classification of T. pallidum as a microaerophilic organism and provide direction for the design of potentially successful culture systems, with or without tissue culture cells.


Streptococcus mutans dextransucrase: stimulation by phospholipids from human sera and oral fluids. Serum, gingival crevicular fluid, and parotid, submandibular, and labial minor gland saliva from four individuals stimulated glucan formation from sucrose by the Streptococcus mutans strain 6715 dextransucrase (EC 2.4.1.5). At final dilutions of 1:10 all of the fluids stimulated crude enzyme preparations approximately 1.8-fold. The fluids stimulated the purified water-insoluble glucan-synthesizing form of the dextransucrase approximately 3.2-fold and the water-soluble glucan-producing form of the enzyme approximately 2.4-fold. The fluids all contained concentrations of stimulatory material that could be reduced to undetectable levels only after dilutions of greater than 1:1,000. The increased rates of glucan formation caused by the fluids and dextran were additive, indicating that stimulation by the fluids was primarily due to interactions with entities other than glucan primer molecules. In contrast, the elevated levels of glucan formation in the presence of the fluids was not further enhanced by the addition of lysophosphatidylcholine. Lysophosphatidylcholine purified from parotid and submandibular saliva by solvent extraction and thin-layer chromatography stimulated the dextransucrase as effectively as egg yolk lysophosphatidylcholine. Thus, phospholipids normally found in human oral fluids can enhance the activity of an enzyme believed to be directly associated with the cariogenic potential of S. mutans.


Genetics of Treponema: characterization of Treponema hyodysenteriae and its relationship to Treponema pallidum. Saturation reassociation assays with 125I-labeled treponemal DNAs show that Treponema hyodysenteriae is genetically unrelated to T. pallidum (Nichols), T. phagedenis biotype Reiter, and T. refringens biotype Noguchi. Pathogenic and nonpathogenic isolates of T. hyodysenteriae exhibited 28% sequence homology and had an extremely low guanine-plus-cytosine content (25.8%).


Intestinal colonization and virulence of Salmonella in mice. Within 3 h after oral challenge of mice with Salmonella typhimurium, foci of infection developed in the Peyer's patches of the small intestine. The numbers of organisms in the cecum, although in excess of those found in the small intestine, were not firmly associated with the cecal wall but were present largely in the cecum's contents. The Peyer's patches at first were remarkably incapable of eliminating even small numbers of Salmonella, but at about 7 days after infection developed the ability to eliminate a less virulent strain of S. typhimurium. Selected strains of Salmonella of varied virulence, and hybrid Escherichia coli/Salmonella typhimurium with varied O-antigens, revealed that those of low virulence could multiply within the intestinal Peyer's patches at nearly the same rate as a virulent strain, and the ability to multiply within the Peyer's patches was not dependent upon O-antigen type or smooth lipopolysaccharide. The ability of these strains to adhere to intestinal mucosa in vitro did not reflect on their ability to colonize the Peyer's patches, although strains of high in vitro adhesive ability appeared in greater numbers initially after oral challenge. Anti-O serum, ineffective in reducing the in vitro adhesive ability of virulent S. typhimurium, when given with the oral challenge prevented Peyer's patch colonization but was unable to prevent the appearance of a systemic infection. Anti-H serum, although effective in vitro in preventing adherence, had no effect in vivo. These experiments suggest that adhesiveness is neither essential nor sufficient for the virulence of Salmonella and that the usual development of a systemic infection after colonization of the small intestinal Peyer's patches may be subverted by the presence of O-antibody.


Immunization of suckling pigs against enterotoxigenic Escherichia coli-induced diarrheal disease by vaccinating dams with purified 987 or K99 pili: protection correlates with pilus homology of vaccine and challenge. Pregnant gilts were vaccinated with purified strain 987 pili (987P), strain K99 pili, or a saline-formaldehyde control. Suckling pigs born to vaccinated gilts were allowed to consume colostrum and were then challenged intragastrically with one of three enterotoxigenic Escherichia coli strains: 987 (O9:K103, 987P:NM), 74-5208 (02O:K101, 987P:NM) or 431 (O101:K30, 99:NM). In litters where the dam was vaccinated with the same pilus as that possessed by the challenge organism, the incidence and duration of diarrhea and the degree of intestinal colonization (either duration or extent) were less than those of the other vaccine groups. Surviving pigs in the homologous vaccine groups also had better weight gains than pigs in the other vaccine groups. The experiments extend and confirm previous reports that vaccination of the dam with purified pili confers protection to neonatal suckling pigs against diarrheal disease caused by enterotoxigenic E. coli strains that possess the same pili. Protection did not extend to enterotoxigenic strains possessing different pili.


Host defenses in murine malaria: induction of a protracted state of immunity with a formalin-killed Plasmodium berghei blood parasite vaccine. Random-bred mice were immunized with a nonliving antigen prepared from mixed-blood forms of Plasmodium berghei, strain NYU-2, in combination with Corynebacterium parvum and/or living BCG. A high proportion of intravenously immunized mice survived virulent challenge, but subcutaneous vaccination was less effective. Vaccinated mice developed a patent infection after challenge similar to that observed in normal controls. However, between days 12 to 20 postchallenge, infections in some vaccinated mice became subpatent, whereas infections in all normal controls progressed until death. The incidence of recrudescent infection was low and, eventually, a state of sterile immunity was established. The capacity of vaccinated mice to withstand P. berghei challenge was sustained at a fairly stable level for the 6-month period of observation. Mice that had survived a primary infection with P. berghei almost completely suppressed a second and larger challenge with the same organism.


Ultrastructural studies on the interaction between Salmonella typhimurium 395 M and HeLa cells. The interaction of Salmonella typhimurium 395 MS and its rough Rd-mutant 395 MR10 with HeLa cells was studied by transmission and scanning electron microscopy. The bacteria attached to central as well as more marginal positions of the HeLa cell surface. Bacteria associated preferentially to HeLa cells with a relatively low number of microvilli, in which they often were entangled. Bacteria attached to the cell border were sometimes surrounded by membrane folds, possibly as a response to their attachment. Infected cells had longer and more slender microvilli compared with noninfected cells. Some parts of the attached bacteria were in close contact with the HeLa cell membrane, whereas other parts were separated from the latter by a gap. Bacteria adhered preferentially to microvilli without obvious membrane damage. Most of the intracellular bacteria were surrounded by a membrane, often appearing as a vacuole, which sometimes contained more than one bacterium. Intracellular bacteria seemed to be morphologically intact. We propose that S. typhimurium enter HeLa cells by a process of phagocytosis.


Lung response to congenitally athymic (nude), heterozygous, and Swiss Webster mice to aerogenic and intranasal infection by Nocardia asteroides. Congenitally athymic (nude, Nu/Nu), heterozygous (Nu/+), and Swiss Webster mice were exposed to virulent Nocardia asteroides GUH-2 inhaled from aerosols or administered intranasally. Clearance of the bacteria from the lungs was determined at 6 h and 1, 2, 3 and 7 days after infection. N. asteroides aspirated into the lungs from intranasal administration were killed less rapidly and induced more severe pulmonary infections than did comparable numbers of organisms inhaled from aerosols. Bacterial clearance and histological data indicated that nude mice were significantly more susceptible to nocardial infection than were heterozygous littermates or Swiss Webster mice. From these data we conclude that: (i) pulmonary defenses cope less well with intranasally administered N. asteroides than with aerosolized organisms, (ii) alveolar macrophages alone appear not to be an efficient barrier to nocardial infections, and (iii) T cells are important to pulmonary clearance and prevention of dissemination of N. asteroides from the lung.


Swiss and inbred mice in the infant mouse test for the assay of Escherichia coli thermostable enterotoxin. Swiss and inbred mice (C3H, ASn, B10A, and C57) were used to assay different thermostable enterotoxins prepared from Escherichia coli strains 3 (human), E57 (swine), B41 (cattle), and S13 (sheep). All strains of mice detected enterotoxins, but some reacted better to a given thermostable enterotoxin preparation than other inbred strains. Since the results obtained with Swiss mice were more consistent than with inbred mice, the former are apparently more suitable in the infant mouse test.


[A new influenza subunit vaccine: hemagglutinating antibodies one year after vaccination (author's transl)]. The antibody response to a new influenza subunit vaccine was compare d one year after vaccination with the responses induced by two other influenza vaccines. The subunit vaccine was given either in a high dose form containing 2100 IU, or in a low dose form containing 700 IU. As comparison a split vaccine was used containing 800 IU and AI(OH)3 as adjuvant and a whole virus vaccine containing 2100 IU. Of the 399 vaccinated subjects which had taken part in this study 151 were available for hemagglutination inhibiting (HAI) antibody determinations one year after vaccination. Protection rates assessed for the respective groups on the assumption that serum HAI titers of 1 : 32 or greater confer protection. With the high dose of subunit vaccine 85% of volunteers were considered still to have protective titers one year after vaccination, compared with 77% of those who received the whole virus vaccine. Although the high dose subunit vaccine and whole virus vaccine induced similarly high protective levels lasting at least one year, the reactions observed on vaccination were significantly less with the subunit preparation. The lower dose of subunit vaccine induced lower levels of protection (60%) after one year, and lower mean HAI titers than the high dose subunit vaccine. Nevertheless protection was superior to that of the split virus adjuvant vaccine. The addition of adjuvant thus does not seem materially to improve the immune response to influenza virus antigens. An increase of antigen content can however be seen as a practical alternative for achieving higher antibody levels. The subunit vaccine would appear to be particularly suitable in this respect as even with a higher dose there is no increase in reactogenicity.


[Vaccination of infants and schoolchildren with an influenza subunit vaccine (author's transl)]. A new influenza subunit vaccine which contains only hemagglutinin and neuraminidase antigens was investigated for reactogenicity and immunogenicity in children aged between three and 15 years. Children under six years of age received either 500 IU or 1000 IU of the commercial vaccine, those aged from six to 15 years either 1000 IU or 2000 IU. The vaccines contained the virus strains recommended by the World Health Organisation for the vaccination season 1976/77. In a double blind study the vaccinees were allocated at random to the different dosage groups. The children were examined for reactions by the vaccinating physician 24 hours after vaccination. Serum hemagglutination inhibiting antibody titers were determined before vaccination and four weeks after vaccination. In the younger age-group additional antibody determination was made two weeks after a booster injection. A very low rate of side-reactions was observed in all dosage groups. The increase of the antigen content was not associated with a higher rate of side reactions. After the first vaccination a significant rise of antibody titers could be observed in all children. After the booster injection a further increase of these antibody titers was observed. The response of the younger age group to the dosages 500 and 100 IU did not different significantly. In contrast, in the older age group the increase of the dosage from 1000 to 2000 IU was connected with a better immune response. This was especially marked in the antibody titers against the influenza B-strain virus.


Therapy of serious infections with cefamandole. Forty-four patients with serious bacterial infections were treated with cefamandole in a dose 1--2 g every four to six hours. Thirty-two patients were cured and six were markedly improved. Three of six failures were due to superinfection with cephalothin-resistant microorganisms. The over-all bacteriologic response was 80%. In 12 of 13 patients with bacteremia the blood was sterilized. Ten of 14 patients with gram-negative bacillary infections responded to treatment. Six of these were due to cephalothin-resistant microorganisms, three of which responded. Fifteen patients who were treated had a history of penicillin allergy. There were no serious reactions although skin rash did develop. Phlebitis was uncommon.


The influence of maternal immunoglobulin-G-antibodies on indirect haemagglutination in newborns. High antibody titers against Escherichia coli were found in 100 randomly selected pregnant women by means of indirect haemagglutination (IHA). After birth, sera from the umbilical vein of the newborns were also tested. It was found that the indirect haemagglutination titers of the children were strongly influenced by the IgG which had been transferred via the placenta. Twenty-two sera of newborns showed an increased IHA-titer against E. coli, but only seven children displayed specific IgM antibodies, determined by indirect immunofluorescence. After chromatographic separation of the sera into IgG and IgM fractions, it was demonstrated that isolated antibodies of the IgG type alone can produce increased IHA titers. The assumption that the indirect haemagglutination is determined almost exclusively by antibodies of the IgG type is unfounded. Therefore, indirect haemagglutination is not suitable as a screening-test for newborns.


Impression technique for denture reconstruction. The advent of tissue conditioning materials has made possible a new method for recording functional impressions for denture reconstruction using the patient's existing dentures as impression trays. The criteria by which the suitability of existing dentures for use in the technique are set out in detail. A particular feature of the method is the physiological recording of the post palatal seal area using Iowa wax and its incorporation into the entire maxillary impression. The advantages of the technique include the possibility of recording the tissues under physiological displacement resulting from normal function over a prolonged period rather than at the moment of final setting of conventional impression materials.


Reactive antibodies in the bronchial washings of lung cancer patients. Lung-carcinoma-reactive antibodies have been previously isolated from tumor tissues and pleural effusions. To explore the immunoglobulin (Ig) content of bronchial secretions, bronchial washings of patients with inflammatory and neoplastic lung diseases were studied with respect to Ig levels and specificity. Expressed as Ig/potassium ratios, statistically significant increases in Ig levels were found in inflammatory diseases and even higher increases in lung carcinomas. The isolation of Ig from bronchial washings was achieved by dissociation of immune complexes at low pH, neutralization and subsequent purification by anion exchange chromatography. The isolated immunoglobulins were tested in indirect immunofluorescence against lung cancer cells of various histologic types in tissue cultures and fresh suspensions. Positive cytoplasmic fluorescence was obtained with cells of adeno carcinomas and squamous-cell carcinomas of the lung but not with cells of normal lung. The accessibility of bronchial washings makes the investigation of their lung-cancer-reactive immunoglobulins relatively easy and raises the possibility of its eventual conversion into a screening test.


Tumour angiogenesis factor (TAF) and its neutralisation by a xenogeneic antiserum. Tumour angiogenesis factor (TAF) has been extracted from a variety of animal and human tumours. Its ability to induce neovascularization in two bioassays, viz. rat air-sac and chick chorioallantoic membrane, is demonstrated. Similar extracts from normal tissues fail to induce angiogenesis. TAF extracts have been used to raise a xenogeneic antiserum, the specificity of which was investigated using classical immunological methods. Its ability to neutralize the biological activity of TAF using the two in vivo assays, was also demonstrated.


Drug use and abuse among women: an overview. A review of the current literature and research on women's usage of three popular drug categories--heroin, marijuana, and the psychotropics--reveals that use and abuse of these drugs by women have been largely neglected in both research and treatment. Women's usage of these three drug categories bears investigation because use and possibly abuse, by women, of all three of these categories is increasing. Heroin addiction among women has increased at a much faster rate than among men, marijuana usage is estimated to equal that of men in some regions of the United States, and use of the psychotropics is consistently higher among women than among men. In all areas of female drug usage, a scarcity of data exists, with some studies including only male respondents. In still other studies, the female user is treated as a deviant user because she does not fit the conceptual models derived from studies of male abusers.


The avowal of motives for marijuana behavior. The present report is a comparative analysis of data from different studies on the motives avowed for the use and nonuse of marijuana. Qualitative data on the reasons given by users and nonusers for their behavior were first described and categorized according to 21 verbal patterns. Then these categories were utilized to analyze quantitative data by various investigators, i.e., were the means to compare the frequencies with which each kind of motive was verbalized. The results indicated that knowledge-ableness is by far the most frequently expressed motive for the initial use of marijuana. Continued use, however, is largely explained either on the grounds of self-fulfillment or with an appeal to psychological drives. Habitual users in different studies mentioned almost all categories of reasons for smoking more often than did occasional users. Explanations for the two types of nonuse were also quite dissimilar. For the discontinued use of pot, lack of interest is claimed as the chief motivation; but for never use, appeals to illegality, injury, drug addiction, and morality are asserted more frequently. Several conclusions based on these findings are drawn.


Chlorosis and chronic disease in 19th-century Britain: the social constitution of somatic illness in a capitalist society. This paper is informed by the recent attempts to construct Marxist views of nature, by the lively historical work on 19th-century British class structure, and by the dedicated work of feminist historians of medicine. Using the common disease of adolescent girls, chlorosis, as an example of a real "physical" illness, I shall argue that disease is, in part, socially constructed. Not only is the social class of both doctor and patient an important determinant in the perception of illness, but so too is the relationship between the disease and the mode of production. Both the "existence" of chlorosis and the way it was understood served ideologically to conceal the growing importance of adolescent labor and the recognition of the social genesis of illness. In doing so, chlorosis was similar to other forms of chronic illness. In a time when the conditions of work were strikingly insalubrious, the etiological emphasis was on individual failure, not on physical or social conditions of work. I argue that notions of health and disease partake of the struggles and social relations of the society that sustains them, but in a way which hides that very social nature. In this sense, they are like Marx's concept of a commodity--and in being a commodity, diseases appear not to embody social relations, but rather to be part of nature. I suggest that we see this "nature" in part as a commodity fetish--something we construct as "other" for a reason--and that we rediscover the social in the natural.


Viminol analgesic activity in elderly patients with chronic pain: a controlled evaluation, using self-rating questionnaires. The analgesic effects of single oral doses of viminol (70 mg), of viminol (70 mg) combined with chlordiazepoxide (10 mg) and of chlordiazepoxide (10 mg) were compared in 41 outpatients with chronic pain mostly due to neoplasms. Alone, or combined with chlordiazepoxide, viminol proved significantly more effective than chlordiazepoxide; No difference in analgestic effect was observed between viminol combined with chlordiazepoxide and viminol alone. The use of self-administered questionnaires was found particularly convenient for study of analgesics in outpatients.


Influence of chenodeoxycholic acid on serum triglycerides in patients with primary hypertriglyceridemia. Administration of chenodeoxycholic acid (CDCA) to patients with primary hypertriglyceridemia caused the diminution of serum triglyceride concentrations of 54.31% and 46.04% at dosages of 500 mg/day and of 250 mg/day, respectively. Results after 1 month of treatment with 250 mg/day are not statistically different from those obtained with 500 mg/day. Administration of CDCA did not change the serum cholesterol, SGOT, SGPT, total bilirubin, gammaGT or alkaline phosphatase levels.


Electron microscopic observations of intracytoplasmic membranous structures in Mycobacterium leprae by means of serial ultrathin sectioning. The fine structures and interconnections between the cytoplasmic membrane and mesosomes of Mycobacterium leprae in human skin were studied in ultrathin sections. These intracellular membranous organelles were seen as laminated structures and as clusters of vesicles, which were trilaminar consisting of two electron-dense layers separated by an electron-transparent zone. The formation of mesosomes seems to be initiated by invagination and/or folding of the cytoplasmic membrane.


Hypotriglyceridemic effect of chenodeoxycholic acid after a short time of administration. Administration of chenodeoxycholic acid (CDCA), 200 mg/day, for 5 days to patients with primary hypertriglyceridemia reduced by 34% the serum triglycerides concentration. Reduction after CDCA is statistically significant vs. the placebo. Even after a short time of CDCA administration, the pharmacological effect is clearly evident.


In vitro cultivation of mycobacteria in cholesterol lecithin media from lepromas of rats infected with Mycobacterium lepraemurium. In vivo grown M. lepraemurium suspensions were inoculated into a basal medium containing cholesterol and lecithin. Slow growing strains of mycobacteria were cultured regularly in these media. The presence of free cholesterol or cholesterol in serum or cholesterol in trypsin-digested egg yolk was essential for growth. The primary cultures were difficult to obtain, but the strains were easily subcultured. A heavy inoculum was necessary to obtain primary cultures in the liquid media, no growth occurred on semisolid agar slants. Similarly slow-growing primary cultures were obtained on Ogawa egg yolk media. Growth developed in a considerably shorter time if Ogawa's medium was enriched with 0.4% yeast extract (Difco). The cultures obtained on Ogawa egg yolk media were successfully subcultered in liquid cholesterol-lecithin media. The relation of the cultured strains of mycobacteria to the pathology of murine leprosy is not yet clear. The dynamics of cholesterol metabolism in the macrophages related to murine leprosy is discussed.


Analysis of a certain type of 2 x 3 tables, exemplified by biopsy findings in a controlled clinical trial. A procedure is described for the evaluation of 2 X 3 Tables, in which the two rows correspond to the two treatments to be compared and the three columns are ordered classifications ("improved", "unchanged", "deteriorated"). The method is illustrated by an example taken from the recent literature: liver biopsy data from a controlled clinical trial of silymarin in the treatment of chronic hepatitis.


The renin-aldosterone system in dihydralazine therapy during hypertensive pregnancy. Ten patients with hypertensive pregnancy were studied in hospital for 3 weeks. Plasma renin activity (PRA), the daily urinary excretion of aldosterone (dU-Aldo) and electrolytes were studied before and during dihydralazine therapy (25 mg 3 times daily). In one week the supine blood pressure decreased (p less than 0.05) without any discernible tachycardia. PRA increased in 3 days of the treatment by 50% (p less than 0.05) and this mean level was maintained. In dU-Aldo no change was found, however, during the study; and the excretions of sodium and potassium remained unchanged as well. In patients without diuretic therapy the basal PRA level before dihydralazine therapy was lower than in normal pregnancy. Also, in patients with preceding diuretic therapy the basal level of PRA was lower than during diuretics in gravidas without hypertension. Judging from the animal experiments, the rise in PRA might be seen as favorable for uteroplacental circulation in pregnancy hypertension.


Dose-response study of a new oral bronchodilator, NAB 365 (clenbuterol). A new oral bronchodilator, NAB 365 (clenbuterol), at doses of 10, 20, 30 and 40 microgram, was studied in a double-blind placebo-controlled incomplete cross-over study in 10 patients suffering from chronic bronchitis who had reversible airway obstruction. The ventilatory response was assessed by measurement of the forced expiratory volume in 1 sec (FEV1), total airway resistance (Raw) and specific airway conductance (sGaw) before and 30, 60, 120, 180, 240, 360 and 480 min after administration. With the 30 and 40 microgram doses, a significant bronchodilating effect was seen between 30 and 360 min after administration, with peak effects between 120 and 240 min. The maximal effects in ventilatory response after 30 microg were only slightly less than after 40 microgram; the bronchodilating effect of 20 microgram was statistically less marked than those of 30 of 30 and 40 microgram on FEV1, but not statistically different on Raw and sGaw. These findings have been briefly discussed. The effects of 10 microgram and of the placebo were always statistically lower than those of 20 microgram. No significant cardiovascular effects and no subjective side effects were observed. In conclusion, the results suggest that 30 microgram of NAB 365 (clenbuterol) are the suitable acute dose.


Reduction of indomethacin-induced gastrointestinal blood loss by sodium salicylate in man. The gastrointestinal bleeding during treatment with indomethacin, aspirin, phenylbutazone and sodium salicylate in healthy human subjects was determined. Gastrointestinal blood loss was determined by the Cr51-labelled erythrocyte method. The method was modified by us. Collected stool was incinerated and radioactivity of ashes was determined. Furthermore, rheumatic patients were treated for 4 weeks with indomethacin alone or with indomethacin combined with sodium salicylate. The gastrointestinal blood loss was determined on the last 4 days of treatment. It was found that blood loss was significantly reduced by combined treatment in comparison to monotherapy with indomethacin, while combined treatment had a marked anti-rheumatic effect.


Some aspects of malignant tumors of the lymph nodes in Iran. Malignant tumors of the lymph nodes are very widespread in Iran. Out of 15,792 lymph nodes examined during a period of 25 years, in 22% of the cases, the diagnosis was a primary malignancy, and in 10% we had metastatic deposits. Out of 40,690 cancer cases diagnosed in pathology laboratories in Tehran, 8.4% were primary malignant tumors of lymph nodes, with a sex ratio of 3:1. The anatomical distribution of malignant lymph nodes shows 46% in the head and neck, with 17% deep in mediastinal or mesenteric areas. The histological typing of these malignancies shows 40% to be Hodgkin's disease, and 60% sarcoma. The distribution by age group shows that these malignant tumors are observed in patients of all age groups, the highest frequency being found between 20 and 54. The statistical data collected in four provinces of Iran, from North to South, reveal that these malignant tumors are observed with the same frequency in other parts of Iran.


Recurrences in primarily infected and uninfected urolithiasis. An analysis was carried out of 640 patients treated because of urolithiasis at the Urological Clinic of the Medical Academy in Warsaw in the years 1971--1974. Factors conducive to stone formation were analyzed. In infected urolithiasis, different bacterial floras were found in men and women. The importance of infections acquired in hospital by patients is emphasized. In primarily uninfected urolithiasis there were 35 per cent recurrences, as against 53 per cent recurrences in infected urolithiasis.


Description and interpretation of functional changes in canine kidneys after preservation with hypothermic perfusates and subsequent reperfusion. The function of canine kidneys stored with hypothermic solutions (Rheomacrodex, Collins4, Sacks) was studied in acute autologous reperfusion models. Changes in the perfusates (increase in LDH level and minor increase in K+ level), as well as the quantitative and qualitative properties (proteinuria) of the urine produced by the graft may be ascribed to storage and revascularization which affect the cells and their membranes. The alterations of the circulatory parameters demonstrable at the outset and during revascularization are due to an increase in renovascular resistance of hypothermic origin on the one hand, and to acute tubular epithelial damage inherent in the procedure, on the other.


A highly sensitive method for the demonstration of carcinoembryonic antigen in normal and neoplastic colonic tissue. A highly sensitive method for the immuno-histochemical localisation of carcinoembryonic antigen (CEA) is described. This method is based on the binding of a peroxidase-antiperoxidase complex (PAP) to anti-CEA antibodies by means of an anti-gamma-globulin which reacts with both the anti-CEA and the antiperoxidase antibodies. Using the technique described here, CEA was localised in conventionally processed normal and cancerous colonic tissue. In normal as well as in neoplastic tissues, a CEA-specific staining of cell membranes and cytoplasm was demonstrated. In frozen sections of normal colonic tissue CEA was found even at high dilutions of the first antibody; this indicates the high sensitivity of the method. The applicability of the method to conventionally processed and thereby well preserved tissue specimens opens the possibility to identify CEA by light microscopy even at very low concentrations.


Distribution of gastrin and CCK cells in the rat gastrointestinal tract. Evidence for the occurrence of three distinct cell types storing COOH-terminal gastrin immunoreactivity. Gastrin and cholecystokinin (CCH) cells of the rat gastrointestinal tract have been studied by immunocytochemistry and radioimmunoanalysis. With antisera directed against the COOH-terminal tetrapeptide sequence, which is common to gastrin and CCK, three distinct endocrine cell types are detected. One of the cell types predominates in the antrum, is scarce in the rest of the gut and corresponds to the gastrin cell. The second cell type is virtually confined to the duodenum and jejunum and corresponds to the CCK cell. The third cell type occurs disseminated in the small intestines, predominates in the ileum, and reacts with COOH-terminus-specific antisera only following diethylpyrocarbonate and not following formaldehyde fixation. It is possible that the third cell type stores a third member of the gastrin-CCK family of gut hormones.


[Surgical correction of mandibular cyst and tumour defects (author's transl)]. The various surgical techniques for the repair of mandibular defects following ablative surgery are discussed. The methods used in the treatment of large mandibular cysts are described and a modification of the osteoplastic technique is presented. The importance of reconstruction and functional rehabilitation with so caused mandibular defects and temporo-mandibular joint substitutions are stressed.


Localization of somatostatin-like immunoreactivity in the Golgi apparatus of central and peripheral neurons. With the indirect immunofluorescence technique of Coons and collaborators somatostatin-like immunoreactivity (SLI) was observed in certain neurons of the central and peripheral nervous system of the rat. In the cell bodies a strong SLI was observed with a distribution resembling that of the Golgi apparatus. In addition a weak SLI was diffusely distributed in the cytoplasm. After photography the sections processed for immunocytochemistry were stained with the thiamine pyrophosphatase technique of Novikoff and Goldfischer. The latter technique is assumed to be a specific marker for the Golgi complex. It was found that the strong SLI and the thiamine pyrophosphatase activity had an identical distribution. Thus, one pool of somatostatin appears to be localized to the Golgi apparatus.


Human alveolar macrophages: antigen-independent binding of lymphocytes. To characterize the initial step in alveolar macrophage (AM)-lymphocyte (L) interaction in the human lung, we studied the ability of human AM to bind autologous blood L in vitro in the absence of antigen. AM were obtained by saline bronchial lavage through a fiberoptic bronchoscope. Monolayers of AM attached to glass bound autologous blood L prepared by Ficoll-Hypaque and nylon wool column separation. The AM-L binding increased from zero time to a maximum at 2 h and then declined to a zero time value at 18 h. The binding was dependent on the number of L added to the AM monolayers, with greatest binding at an AM:L ratio of 1:50. AM:L binding required viability of AM, but not of L, and was temperature dependent. Pretreatment of AM with iodoacetic acid, trypsin, neuraminidase, or colchicine diminished attachment of L at 2 h. Neuraminidase pretreatment of L resulted in increased binding to nontreated AM. Thus, a physical interaction between human AM and autologous peripheral blood L can occur in vitro in the absence of known antigen; similar interaction in vivo may play a role in the generation of cell-mediated immune responses in the lung.


Effect of histamine and methacholine on guinea pig tracheal permeability to HRP. The effects of histamine, methacholine, and ether on the permeability of the respiratory mucosa to macromolecules were investigated employing a radioimmunoassay and histochemical techniques to monitor movement of horseradish peroxidase (HRP) from airway lumen to blood. We found that 0.08% of the dose of HRP instilled into guinea pig tracheas was present in the blood volume at 10 min, and plasma HRP levels increased at a rate of 0.0036% instilled dose/min thereafter. After inhalation challenge, significant increases in plasma rates of accumulation of HRP were recorded for the histamine, methacholine, and ether groups, whereas no change in rate was noted for the control (Tyrode's) group. Electron micrographs of tracheal sections showed HRP penetration into the intercellular spaces of the epithelium after histamine, methacoline, or ether exposure but no penetration in the Tyrode's group. We conclude that, like ether, histamine and methacholine increase tracheobronchial permeability and this effect is most likely mediated by a functional change in the epithelial tight junction.


The functional relationship between artificial food colors and hyperactivity. The presence of a functional relationship between the ingestion of artificial food colors and an increase in the frequency and/or duration of selected behaviors that are representative of the hyperactive behavior syndrome was experimentally investigated. Two eight-year-old females, who had been on the Feingold K-P diet for a minimum of 11 months, were the subjects studied. The experimental design was a variation of the BAB design, with double-blind conditions. This design allowed an experimental analysis of the placebo phases as well as challenge phases. Data were obtained by trained observers on Out of Seat, On Task, and Physically Aggressive behaviors, as they occurred in the subjects' regular class setting. Results indicated (a) the existence of a functional relationship between the ingestion of artificial food colors and an increase in both the duration and frequency of hyperactive behaviors, (b) the absence of a placebo effect, and (c) differential sensitivity of the dependent variables to the challenge effects.


Relaxation exercise with cerebral palsied adults showing spasticity. A method for improving the muscle incoordination associated with cerebral palsy was empirically tested. Relaxation exercises, involving the successive tensing and relaxing of the major muscle groups of the body, were performed by four adults with spastic quadraplegia. All four subjects were sheltered workshop employees. Their IQs ranged from normal to mentally deficient. Testing consisted of two timed-trials of both the Placing Test and the Turning Test from the Minnesota Rate of Manipulation Tests. These tests measured the amount of time required to complete various manual manipulations involving pegs and pegboard. A multiple baseline across subjects design was employed to provide experimental control. All subjects showed significant improvement over baseline performance on the tests following the relaxation training. Limitations of this preliminary investigation and implications of these results for future research were noted.


Comparison of two rapid test procedures with the standard EC test recovery of fecal coliform bacteria from shellfish-growing waters. A study was conducted to compare recovery and enumeration capability of two 24-hr multitube fermentation tests with the standard EC test for fecal coliform levels in shellfish-growing waters. The 2 tests were the A-1 test developed by Andrews and Presnell, specifying 24-hr incubation in A-1 medium at 44.5 degree C; and a modification of the A-1 test requiring a 3-hr resuscitation of 35 degree C before incubation at 44.5 degree C for 21 hr. Fifteen State, Federal, and Provincial laboratories examined 10 routine shellfish-growing area samples per month in parallel by the 3 methods for 1 year. IMViC tests (indole, methyl red, Voges-Proskauer, and citrate) were conducted on all gas-positive tubes. The modified A-1 test recovered higher levels of fecal coliforms than the A-1 test. Although there were seasonal and geographic variations in recovery and enumeration by the modified A-1 test, overall there was good correlation of the modified A-1 test with the EC test. Both the A-1 and modified A-1 tests were more specific for Escherichia coli than the EC test. Results of the study indicate that the 24-hr modified A-1 test can be used as an alterantive test for the standard 72-hr EC test as an adjunct to the sanitary survey for the classification and control of shellfish-growing areas waters.


A controlled crossover study of triiodothyronine in autistic children. A placebo-controlled crossover study of behavioral effects of triiodothyronine (T3) was conducted in 30 young clinically euthyroid autistic children. Multiple independent raters and multiple rating scales were used. Except for a few symptoms that were reduced on T3, the drug did not differ from placebo. Time itself accounted for most of the improvement in the whole sample. As a group, the lower IQ children responded to T3. The individual children who were responders could not be defined by any parameter.


Role of uracil-DNA glycosylase in the repair of deaminated cytosine residues of DNA in Escherichia coli. Uracil-DNA glycosylase, which acts specifically on uracil-containing DNA, was purified 250-fold from an extract of Escherichia coli 1100. The enzyme releases free uracil from DNA, producing alkali-labile apyrimidinic sites in the DNA. The enzyme is active on both native and heat-denatured DNA of phage PBS1, which contains uracil in place of thymine. piX174 DNA which had been treated with bisulfite and then at alkaline pH was susceptible to the action of uracil-DNA glycosylase. Since DNA treated with bisulfite alone was less susceptible to the enzyme, it is likely that the enzyme recognizes deaminated cytosine, namely uracil, but not bisulfite adducts of uracil and cytosine in the treated DNA. DNA treated with nitrite or hydroxylamine was not attacked by the enzyme. Enzyme activity acting on bisulfite-treated DNA was absent from an extract of E. coli mutant BD10 (ung). The mutant exhibited higher sensitivity to bisulfite than did the wild-type strain and was unable to reactivate phage T1 pre-exposed to bisulfite and weak alkali.


An enzyme activity specific for nitrous acid-treated DNA in Escherichia coli. An enzyme activity specifically active on nitrous acid-treated DNA was found in an extract of Escherichia coli. The enzyme acts on both double- and single-stranded DNAs, treated with nitrous acid, in the presence of EDTA, although the former DNA is a better substrate. Evidence is presented that nitrous acid- and bisulfite-induced types of damage in DNA are recognized by different enzymes: (1) Uracil-DNA glycosylase, purified 250-fold from E. coli 1100, attacks bisulfite-treated DNA but not nitrous acid-treated DNA. (2) Almost equal levels of activity toward nitrous acid-treated DNA were found in wild-type and uracil-DNA glycosylase-deficient strains of E. coli.


Total hip replacement in congenital dislocation and dysplasia of the hip. The results of thirty-one total hip replacements in twenty-four patients with either severe congenital dysplasia or dislocation, after an average follow-up of four years, were excellent in eleven, good in sixteen, fair in one, and poor in one. The operative technique included superolateral bone grafts to increase the acetabular coverage in six hips. Twenty-seven hips required smaller and straighter femoral components than normal. The incidence of major complications was 19 per cent.


Distraction-lengthening of digital rays in the management of the injured hand. The distraction-lengthening principle and apparatus, formerly used in the hand for reconstruction in congenital anomalies and traumatic amputations, is described for reconstruction of metacarpals and phalanges shortened by injuries, especially gunshot wound, where those bones have lost significant length. Hands in which single metacarpals have been shortened or in which multiple digits are involved may benefit from the technique.


Microvascular anastomoses for bone grafts in the treatment of massive defects in bone. Six patients with large defects in bone are described in whom we performed microvascular anastomoses of grafted fibular vessels (arteries and veins) to vessels in the recipient site. Two other patients, with massive loss of bone and skin, were treated by grafting of osteocutaneous composites also using microvascular anastomoses. All but one defect healed successfully. There is a wide potential for applications of these two techniques in the treatment of large segmental bone defects secondary to trauma or following tumor resection.


Decoration of specific sites on freeze-fractured membranes. Fracturing under ultrahigh vacuum (UHV, P less than or equal to 10(-9) Torr) produces membrane fracture faces devoid of contamination. Such clean surfaces are a prerequisite for studies of interactions between condensing gases and distinct regions of a surface. For the study of water condensation, a device has been developed which enables production of pure water vapor and controlled variation of its partial pressure in an UHV freeze-fracture apparatus. Experiments with yeast plasmalemma fracture faces, produced at -196 degrees C and exposed to pure water vapor before replication, resulted in a "specific decoration" with ice crystals of those pits in the extraplasmic face where the corresponding particles of the plasmic face had been removed. Because water condenses as discrete ice crystals which resemble intramembrane particles, ice crystals might easily be misinterpreted as actual membrane structures. At low specimen temperature (T less than or equal to 110 degrees C) the structural features of membrane fracture faces produced under high vacuum (P approximately 10(-6) Torr) should, therefore, be interpreted with caution.


Analysis of the distribution of spindle microtubules in the diatom Fragilaria. The spindle of the colonial diatom Fragilaria contains two distinct sets of spindle microtubules (MTs): (a) MTs comprising the central spindle, which is composed of two half-spindles interdigitated to form a region of "overlap"; (b) MTs which radiate laterally from the poles. The central spindles from 28 cells are reconstructed by tracking each MT of the central spindle through consecutive serial sections. Because the colonies of Fragilaria are flat ribbons of contiguous cells (clones), it is possible, by using single ribbons of cells, to compare reconstructed spindles at different mitotic stages with minimal intercellular variability. From these reconstructions we have determined: (a) the changes in distribution of MTs along the spindle during mitosis; (b) the change in the total number of MTs during mitosis; (c) the length of each MT (measured by the number of sections each traverses) at different mitotic stages; (d) the frequency of different classes of MTs (i.e., free, continuous, etc.); (e) the spatial arrangement of MTs from opposite poles in the overlap; (f) the approximate number of MTs, separate from the central spindle, which radiate from each spindle pole. From longitudinal sections of the central spindle, the lengths of the whole spindle, half-spindle, and overlap were measured from 80 cells at different mitotic stages. Numerous sources of error may create inaccuracies in these measurements; these problems are discussed. The central spindle at prophase consists predominantly of continuous MTs (pole to pole). Between late prophase and prometaphase, spindle length increases, and the spindle is transformed into two half-spindles (mainly polar MTs) interdigitated to form the overlap. At late anaphase-telophase, the overlap decreases concurrent with spindle elongation. Our interpretation is that the MTs of the central spindle slide past one another at both late prophase and late anaphase. These changes in MT distribution have the effect of elongating the spindle and are not involved in the poleward movement of the chromosomes. Some aspects of tracking spindle MTs, the interaction of MTs in the overlap, formation of the prophase spindle, and our interpretation of rearrangements of MTs, are discussed.


Localization of actin and microfilament-associated proteins in the microvilli and terminal web of the intestinal brush border by immunofluorescence microscopy. Indirect immunofluorescence microscopy was used to localize microfilament-associated proteins in the brush border of mouse intestinal epithelial cells. As expected, antibodies to actin decorated the microfilaments of the microvilli, giving rise to a very intense fluorescence. By contrast, antibodies to myosin, tropomyosin, filamin, and alpha-actinin did not decorate the microvilli. All these antibodies, however, decorated the terminal web region of the brush border. Myosin, tropomyosin, and alpha-actinin, although present throughout the terminal web, were found to be preferentially located around the periphery of the organelle. Therefore, two classes of microfilamentous structures can be documented in the brush border. First, the highly ordered microfilaments which make up the cores of the microvilli apparently lack the associated proteins. Second, seemingly less-ordered microfilaments are found in the terminal web, in which region the myosin, tropomyosin, filamin and alpha-actinin are located.


Presence of spectrin in untreated Friend erythroleukemic cells. Its accumulation upon treatment of the cells with dimethyl sulfoxide. Friend leukemia cells (FLC) are nucleated erythroid precursors, and are markedly stimulated towards more advanced stages of differentiation by treatment with dimethyl sulfoxide (DMSO). The presence of spectrin, an erythrocyte membrane protein, has been investigated in untreated and in DMSO-treated FLC by indirect immunofluorescence and by analysis in SDS-polyacrylamide gel electrophoresis of low-ionic-strength cell extracts immunoprecipitated with a monospecific anti-spectrin serum. Spectrin is detectable in significant amounts in the "inducible" clones prior to DMSO stimulation, and accumulates 4- to 5-fold upon addition of this compound to the cultures. Spectrin accumulation occurs rather early (24 hours after cell seeding) and reaches its peak on the third day to decline thereafter. Semiquantitative determinations of spectrin amounts present in DMSO-stimulated 745A and A degree 1 cells on the third day after treatment were 2.4 X 10(5) and 3.0 X 10(5) molecules/cell, respectively. Spectrin is also detectable in very low amounts in an "uninducible" line of FLC, and is not accumulated upon DMSO treatment thereof, whereas treatment with hemin does cause a significant increase of spectrin-positive cells. These data indicate that spectrin is a convenient "early" marker for in vitro studies of erythropoiesis.


High performance liquid chromatographic analysis of fermentation broths: cephalosporin C and tylosin. Procedures are described for the analysis of cephalosporin C or tylosin in fermentation broths by high performance liquid chromatography (HPLC). Quantitation of the major components in cephalosporin C broths was done by a system involving separation by an ion-pair technique. Tylosin data were obtained using a reverse phase method. A method of equating uv potency of tylosin to microbiological potency is discussed. A comparison of HPLC with a microbiological method for the determination of tylosin concentration is also made.


Simple assay for staphylococcal enterotoxins A, B, and C: modification of enzyme-linked immunosorbent assay. The enzyme-linked immunosorbent assay (ELISA) introduced for the detection of staphylococcal enterotoxins by Saunders et al., Simon and Terplan, and ourselves has proved to be a simple, reliable, and sensitive test. A new modification is described that uses polystyrene balls (diameter, 6 mm) coated individually with antibody against one of the toxins A, B, or C. In a single tube, 20 ml of the food extract was incubated with the three balls differently stained, which were then each tested for the uptake of enterotoxin by a competitive ELISA. A concentration of 0.1 ng or less of enterotoxin per ml can be measured, making tedious concentration procedures of the extracts superfluous. Culture supernatants and extracts from foods artificially or naturally contaminated with toxin were successfully examined. Cross-reactions did not occur, and nonspecific interfering substances did not create serious problems.


Direct use of counterimmunoelectrophoresis in detection of group B streptococci in specimens containing mixed flora. Isolation procedures for the group B streptococci were bypassed by applying counterimmunoelectrophoresis (CIE) directly to the broth containing the initial inoculum. A total of 458 tests were performed on 389 specimens received from 93 mothers and 81 babies. Of all tests, 19.4% were positive group B streptococci by the Lancefield precipitin method. Twenty hours of incubation were required to produce a positive CIE result in 54% of those specimens which were eventually proven a contain group B streptococci by Lancefield technique. There were no false positive CIE results. It is recommended that the initial broth culture be sampled and tested with CIE after an overnight incubation in those cases where another 24 h may be critical in terms of identifying the group B streptococci. A positive result with CIE can be clinically relied upon, but a negative result is indeterminate, and routine isolation procedures would need to be followed in that case.


Micromethod for biochemical identification of coagulase-negative staphylococci. We have endeavored to elaborate a suitable method for easy and rapid identification in clinical microbiology laboratories of the different species of infection-inducing, coagulase-negative staphylococci. Ten type strains described by Kloos and Schleifer and 269 strains isolated from 95 patients were tested; the classical tests were used for determination of Staphylococcus species. Strains were identified by using the Kloos-Schleifer reference method and the micromethod simultaneously. After preliminary tests on 77 substrates, 19 were retained, 15 for determination of species and 4 to reveal biotypes. The substrates were placed in wells in a rigid strip of inert plastic. Inoculation of wells was carried out with rich microbial suspensions in a special medium; reading of substrate reactions was done after incubation for 48 h at 35 degree C. The intrasystem reproducibility was excellent, from 91 to 100% for the 19 substrates. It was in excellent agreement with the reference method, 100% for type strains and 97.9% for hospital-isolated strains. Because it is simple and easy to reproduce, the micromethod will be most useful in clinical and ecological microbiology laboratories.


Complement fixation antibody test for human nocardiosis. Complement-fixing antibody was detected in 13 of 16 patients with histological and/or culture evidence of infection with Nocardia species. The antigen used was a filtrate of soluble substances secreted into liquid growth medium by cultures of N. asteroides. Apparent false positives reactions were found in three of three patients with leprosy and two of five  patients with tuberculosis--results similar to some previously reported methods. N. asteroides and mycobacteria share antigens. Only the false positives with tuberculosis are considered a diagnostic problem. No reactions were obtained with sera from 26 patients with other infections and 41 unifected individuals. Whereas previous nocardia serodiagnostic methods have a sensitivity of approximately 50%, our overall sensitivity (81%) compares favorably and included 9 of 11 positive tests in immunocompromised nocardiosis patients (a source of false negative reactions with previous methods).


Commercial cryptococcal latex kit: clinical evaluation in a medical center hospital. A commercial latex kit for the detection of cryptococcal antigen (LCAT) was used in a medical center hospital to test cerebrospinal fluid (CSF) and serum specimens from patients suspected of having cryptococcal infections. The LCAT was also performed on 8 CSF and 2 serum specimens from other mycotic infections and on 50 serum specimens from healthy controls. Of a total of 561 specimens (489 CSF and 72 sera) from 426 patients, 78 (13.9%) specimens were LCAT positive; these came from 12 patients with culturally proven cryptococcosis. Eleven of these 12 patients were diagnosed as having disseminated cryptococcosis (9 with meningitis). Fourteen other patients, all with positive cultures for Cryptococcus neoformans but nevative LCATs on sera or CSF, were found not to have disseminated infections. All CSF and sera from other mycotic infections and all 50 control sera were negative. No false positive or flase negative tests were encountered. It was concluded that the commercial kit is useful for diagnosis and prognosis of disseminated cryptococcosis, but not as useful for infections such as localized pulmonary or cutaneous cryptococcosis.


Laboratory identification and epidemiology of streptococcal hospital isolates. A total of 343 streptococcal strains were identified to species on the basis of the progressive method of bacterial identification as advocated by Cowan & Steel's Manual for the Identification of Medical Bacteria. Comparative studies were also performed with these strains to determine the accuracy and feasibility of using various types of blood, differential media, and biochemical tests in conjunction with the progressive method of identification. The streptococcal species were then correlated with the type of specimen, sex, and age of hospitalized patients to obtain some insight into the epidemiology of hospital streptococcal isolates.


The use of a prosthetic speech aid for a laryngectomized esophageal speaker with right hemiparesis of the tongue: a case report. A dental-palatal prosthetic device, which had been demonstrated to aid laryngectomees who use the glossal press method of esophageal speech, was used on a patient whose primary method of air charging was bilabial plosive injection and who had a right hemiparesis of the tongue. Cointervention of a prosthodontist and a speech pathologist is described.


Tooth allografts fail to stimulate immunologic memory in mice. Tooth transplants should not preclude future transplants in the recipient. In this study teeth failed to stimulate immunologic memory in weakly disparate mice. The teeth did carry transplantation antigens and were rejected after second-set skin grafts. The teeth probably enhanced their own survival through some immunoregulatory mechanism.


LETS protein in normal and pathological human oral epithelium. The distribution of LETS protein in human oral mucosa was studied by indirect immunofluorescence. Normal epithelium showed surface staining. Intracellular staining occurred in epithelial cell cytoplasm in lichen planus and pemphigoid. Surface staining was absent in discoid lupus erythematosus. In pemphigus, intercellular staining was seen near areas of acantholysis.


Repigmentation of leukoderma by minigrafts of normally pigmented, autologus skin. A method of repigmenting some leukodermas by transplantation of minigrafts of normally pigmented, autologous skin into them is described. Such grafts in addition to retaining their pigment stimulate repigmentation around them by migration of melanocytes and spread of pigment from the grafts. Three patients, one with piebaldism, another with leukoderma from monobenzyl ether of hydroguinone, and a third with depigmentation following healing of a burn enjoyed successful and cosmetically acceptable repigmentation from practice of the method.


ASSESS: Adjustment Scales for Sociometric Evaluation of Secondary-School Students. The Adjustment Scales for Sociometric Evaluation of Secondary-School Students (ASSESS) was designed to assess peer ratings of the behavior of male and female adolescents. Five scales were developed and cross-validated: Aggression-Disruptiveness, Withdrawal, Anxiety, Social Competence, and Academic Difficulty. Evidence of adequate internal consistency and test-retest reliability was presented. Evaluations by male and female classmates were highly correlated. Sex and age differences accounted for only a small proportion of the total variance.