Prenatal development of central optic pathways in albino rats. The development of the central optic projections in albino rat fetuses has been studied using light and electron microscopic degeneration techniques and the horseradish peroxidase method for demonstrating axonal projections of neurons. The first optic axons to reach the region of the optic chiasm arrive at day 15. By day 16, a substantial optic chiasm is seen and the optic tract can be traced into the epithalamus, having first passed through the ventral lateral geniculate nucleus and a thin lamina of cells which is thought to correspond to part of the future dorsal geniculate nucleus. A growth rate of 80-100 mum per hour is estimated for the fastest growing axons. By day 16-1/3 the first axons have entered the anterior border of the superior colliculus and in the next day have grown across the entire rostrocaudal extent with the exception of the medial and lateral edges. The optic axons are recognized at day 17 as bundles lying just below the surface, but in older animals they come to lie deeper, as the whole layer of optic innervation broadens. The first synapses to be formed in the superior colliculus (some of them of optic origin) appear on day 17. Subsequently, there is a gradual increase in the number of contacts, the great majority being formed by optic axons. Compared with previous studies on Xenopus and chick, one of the most striking features of the development of the central visual connections in the rat is the relatively long time before the first optic axons reach the brain and the speed with which they innervate the central structures once they have arrived.


Chemical composition of Streptococcus mutans type c antigen: comparison to type a, b, and d antigens. Studies of the serotype a, b, and d antigens of S mutans were extended to include the serotype c antigen. Type c antigen was extracted from cells of strain Ingbritt and purified by repeated column chromatography. Two type c antigens fractions were separated by chromatography on DEAE-Sephadex and were designated I and II. On agar gel diffusion, the major components of antigens I and II were serologically identical with the major component of the crude Ingbritt extract. Rhamnose and glucose were present in a 2.4:1 ratio and comprised 98 and 93% of the purified antigens, respectively. The compositions of the c antigens were directly related to the carbohydrate content of the serotype c cell wall. In contrast, purified a, b-II, and d antigens, which are also cell wall polysaccharides contain only trace amounts of rhamnose, a major wall carbohydrate. Studies which used comparative immunoelectrophoresis demonstrated that the purified a, b-I, and d antigens were serologically identical to the antigen preparations that have been used for the immunological classification of these serotypes.


Grouping and cross-reacting antigens of oral lactic acid bacteria. The grouping antigens from five of the seven serological groups of lactobacilli have been defined with respect to the carbohydrate units responsible for serological specificity. When the antigens are polysaccharides, specificity is more absolute than with the teichoic acids where cross-reactions may occur because of the same or similar carbohydrate substituents, or more particularly because of the common "backbone" structure of the glycerol teichoic acids. This glycerol phosphate "backbone" accounts for a number of reports on the presence of a "common antigen" in gram-positive organisms and their culture fluids, with the amount of extracellular material being particularly high for S mutans strains.


Improved salt fractionation of animal serums for immunofluorescence studies. With all animal serums studied except rat serum, an improved salt fraction that contained a high percentage of gamma-globulin and little or no albumin could be obtained by using optimal ammonium sulfate concentrations. These concentrations were less than the routinely used half-saturated solutions and different from the sometimes quoted one-third-saturated solutions. These are simple, economical methods for obtaining the antibody-containing globulin fractions from the serums of a variety of animals commonly used as antibody producers for immunofluorescence applications. Fluorescent antibody reagents prepared in this laboratory from serum fractions obtained by these optimal procedures provided conjugates that may be superior to those prepared from fractions obtained with a higher and uniform salt concentration.


Conjugation methods in immunofluorescence. We have described methods of labeling antibody preparations with FITC, TMRI, and RBI. The degree of labeling with FITC can be precisely controlled by using well-defined conjugation procedures and FITC of a known degree of purity. Our experience shows that relatively high F/P ratios of the order of 20 to 25 mug/mg are desirable for antibacterial conjugates. Many commercial preparations of rhodamine isothiocyanate are of very poor quality and are unsatisfactory for use in conjugate preparation. Therefore, one should analyze the rhodamine isothiocyanate product before preparing immune conjugates. Our experience indicates that very satisfactory conjugates of immune IgG or pure antibody can be prepared with TMRI of about 60% purity by using a dye-protein ratio of 20 mug/mg. The optimal dye-IgG ratio for labeling with RBI appears to be about two times that for labeling with TMRI because of the lower specific absorbance and fluorescence emission of RBI. Rhodamine conjugates may be preferred to FITC conjugates in certain situations where tissue autofluorescence interferes with the observation of the yellow-green emission of FITC. Furthermore, mixed rhodamine and FITC conjugates of different specificity can be used to great advantage in double-staining techniques that allow simultaneous screening for two antigenically different organisms on a single microscope slide.


Physicochemical characterization of direct fluorescent antibody reagents. When the data from performance and physicochemical studies of conjugates are combined for analysis, the performance data and specific titers show a direct relationship to the physicochemical data (Table 2). These reagents were prepared from the same lot of antiserum. The specific titers are very misleading without the accompanying data (Table 2). The protein concentrations range from 4 to 10 mg/ml, the F/P ratios from 10 to 30, and CASE shows gamma-globulin to constitute 30 to 100% of the protein. CASE also shows the gamma-globulin F/P ratio to be only 10 to 20. Using these data, we calculated the concentrations of the gamma-globulins and normalized their titers to 10 mg/ml. The value of good fractionation procedures for recovering gamma-globulin and the desirability of obtaining optimal F/P ratios are reflected in the adjusted titers. Physicochemical characterization of conjugates identifies superior and deficient reagents and frequently reveals the cause of inadequate performance. In this way it serves as a quide for improving reagent quality.


Improvement in specificity of immunofluorescent reagents for identifying Streptococcus mutans by DEAE-cellulose-bacterial cell column immunosorption methods. We have described a new method for preparing bacterial cell columns, in which DEAE-cellulose is used as the cell support material. The columns can be used to isolate pure antibody or to remove cross-reacting antibodies from immune IgG. The columns are remarkable stable, can be scaled up to any desired dimensions, and should be suitable for large-scale production of highly specific serological reagents. The method is practical for obtaining serotype-specific and polyvalent IF reagents for identifying S. mutans organisms. However, the method could also be used as a tool for sophisticated studies of antigenic relationships that exist among these and other taxonomic groups of microorganisms.


Optimum immunization of rabbits for Streptococcus mutans antiserum and conjugate production and studies of batch immunoabsorption methods. By far, the most significant rises in titers were seen with the immunization protocol used in series 6. Conjugates prepared from bleedings on the 33rd day produced exceptionally high titers for type b S mutans, and reasonably high titers for type a were obtained in a short time. A concentrated antigen with Formalin (13.4 ml) was given during a ten-day period followed by a two-week rest period, after which booster doses of either antigen with Formalin or live antigen were given (Fig 1). Based on evaluation of the immunization protocol just described, series 6 resulted in the highest titered reagents, but the data are insufficient to permit recommending that particular schedule without limitations. Our experience in the use of live antigens of S mutans for immunization is limited in that only types b, c, and e have been used in this way. The rabbits survived these injections, but the pathogenicity of other strains and other serotypes has not been determined. In addition, protocols including combined injections of killed and living organisms should be tested further for possible improvement in antibody production. In view of these considerations, our recommendations for production of high titered antiserums for S mutans in rabbits are as follows: -Take a preimmunization bleeding from each rabbit and screen by indirect FA tests with the antigens to be used. -Inject heavy concentrations (40 IU/ml) of Formalin-killed cells, intravenously. -Inject for eight to ten consecutive days, giving increasing doses of antigen ranging from 0.2 to 5.0 ml for a total of 12 to 15 ml. -Rest the rabbits for one week. If you are monitoring the progress of immunization, bleed the rabbits before giving booster injections. -Give booster injections on four consecutive days, giving 0.25, 0.5, 1.0, and 1.5 ml of live antigen that has been washed one time to remove traces of media and adjusted to a concentration of 40 IU/ml. If live antigen is not used, continue to give booster injections with killed antigen, injecting 2.0 ml on each of three consecutive days. -Rest the rabbits for one week and take sufficient blood to produce the trial reagents needed, or exsaguinate the rabbits. Absorption of type a conjugates resulted in the total loss of titer for type a cells. The cross-reactions with type b conjugate were easily eliminated by dilution, with the exception of the cross-reaction with S sanguis JC-43. Bratthall's absorption method eliminated all cross-reactions of the type b conjugate. Absorption of type c conjugate successfully removed the cross-reaction with type e cells; however, the loss of homologous type c titer was so great that this absorption is of limited value. High-titered conjugates for types d and e have been obtained by using batch absorption procedures.


Application of fluorescent antibody methods in the analysis of plaque samples. Direct FA staining for S mutans serotypes may be performed on smears made from plaque or strain isolates and on colonies attached to agar plates of black membrane filters. Staining with single conjugates directed to S mutans serotypes (a to e) as well as with polyvalent a-, b-, d-, f-fluorescein label and c-, e-rhodamine label conjugates indicate that serotype c is the most common. This is in agreement with many reports by other investigators. Cross-reactions with many S mutans conjugates occurred with organisms resembling Lancefield groups C and G streptococci.


Allergy to insect stings. II. Phospholipase A: the major allergen in honeybee venom. In order to determine the proteins of major allergenic importance in honeybee venom (Apis mellifera) it was chromatographed on G-50 Sephadex. The four major protein peaks eluted were identified as hyaluronidase, phospholipase, melittin, and apamin. Testing these preparations on the leukocytes of 6 honeybee-sensitive patients, with the in vitro method of histamine release, revealed that all individuals were most sensitive to phospholipase A. IgE antibodies against phospholipase A (RAST) were found in the sera of honeybee-sensitive patients and IgG antibodies to this venom component were found in the sera from beekeepers and venom-treated patients. Melittin appeared to be allergenic in several patients, but the results were variable and were possibly due to contamination with phospholipase. All patients were insensitive to the hyaluronidase and apamin preparations. We conclude that phospholipase A is the major allergen of honeybee venom and, since this protein is readily available, it should be useful for diagnostic and therapeutic studies as well as for the standardization of materials used in the management of honeybee-sensitive patients.


Heavy chain variable region allotypic sub-specificities of rabbit immunoglobulins. I. Identification of three subpopulations of  a1 IgG molecules. Four anti-al Ab subpopulations were isolated from an anti-al antiserum by sequential immunoadsorption chromatography. These four anti-al Ab subpopulations were differentially bound by two "limited heterogeneity" Abs having different components of the al allotypic specificity. Each of the four anti-al Ab subpopulations reacted with al IgG molecules obtained from a2 and a3 rabbits. A subpopulation designated anti-al Ab reacted with 100% of al IgG molecules. Thus, the anti-al-A Ab recognizes al determinants common to all al IgG molecules. Each of the other three subpopulations, designated anti-al-B Ab, anti-al-C Ab, and anti-al-D Ab, reacted with only a fraction of the al IgG molecules but the sum of the percentages of al IgG molecules which reacted with each of these three anti-al Ab subpopulations approximated 100% of the al IgG molecules. Thus each of the anti-al-B Ab, anti-al-C Ab, and anti-al-D Ab recognizes non-common determinants distinct for each of three subpopulations of al IgG molecules. Although 65 to 90% of IgG molecules in al homozygous rabbits have the al allotypic specificity, these IgG molecules are heterogeneous with respect to their antigenic determinants comprising the al allotype; at least three kinds of al IgG molecules are identified. This heterogeneity probably reflects variation in the amino acid sequence of the Vh region of al IgG molecules and, therefore, poses a similar argument which had led to the hypothesis of two genes for one polypeptide chain and to the theory of episomal insertions for the genetic control of immunoglobulin synthesis.


Suppression and reversal of allergic encephalomyelitis in guinea pigs with a non-encephalitogenic analogue of the tryptophan region of the myelin basic protein. The administration of synthetic peptide S42 leads to suppression and reversal of experimental allergic encephalomyelitis (EAE) induced in guinea pigs by myelin basic protein. Peptide S42 contains a linear sequence of 21 amino acid residues, H-Phe-Ser-Trp-Gln-Lys-Phe-Ser-Trp-Gln-Lys-Phe-Ser-Trp-Gln-Lys-Phe-Ser-Trp-Gln-Lys-Gly-OH, made up of four repeating unit sequences of H-Phe-Ser-Trp-Gln-Lys-OH in addition to a C-terminal glycine. Injected at relatively high doses, peptide S42 is non-encephalitogenic. It induces delayed-type hypersensitivity which is not followed by EAE, and elicits delayed-type hypersensitivity responses in peptide S42, encephalitogenic trytophan peptide, or BP-challenged animals for either of the three antigens. The repeating unit sequence of peptide S42 is analogous to the encephalitogenic tryptophan region of the BP molecules . The sequence homology is responsible for cellular recognition of this antigen by the skin test assay and suggests in vivo interaction between peptide S42 and EAE-inducing cells leading to suppression and reversal of disease.


Antigenic determinants of adenovirus capsids. I. Measurement of antibody cross-reactivity. Evidence is presented that the type-specific antibody to the adenovirus hexon is not simply the antibody with the highest activity for cross-reactive determinants, but is a distinct, minority population that recognizes seperate determinants. To quantify it, we have developed an inhibition method with radio-immunoprecipitation (RIP) as a sensitive assay for the type-specific antibody that remains after all the excess of cross-reactive antibody has been blocked by heterologous antigen. During the primary response, 0.1 to 1% of antibody to types 2 or 5 hexon is type-specific, but after boosting, this population may reach 10 to 20%. Antibody to fiber is more than 70% type-specific during primary and secondary responses. The cross-reacting antibody can be removed on immunoabsorbent columns without affecting the virus neutralization titer of the serum.


Further characterization of the anti-encephalitogenic protein (SCP): isolation from bovine spinal cord and spinal roots. The three molecular forms of the anti-encephalitogenic protein, beta-SCP, gamma-SCP, and SCP-peptide were isolated in higher yield by a shortened procedure, which involved 1) extraction of bovine spinal cord (BSC) or bovine spinal roots (BSR) with 0.05 M sodium acetate buffer, pH 4.5, 2) batch absorption on CM-52 cellulose, 3) stepwise elution with sodium acetate buffers, pH 5.8, containing increasing concentrations of sodium chloride and finally, 4) removal of trace contaminants by gel-exclusion chromatography on Sephadex G-50 superfine. The m.w. of the purified proteins determined by sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis was 13,200 daltons. The same value for the molecular sizes was obtained by gel-exclusion chromatography by using 0.1% SDS in 0.05M sodium chloride as eluant. In the absence of SDS the molecular sizes estimated by gel exclusion chromatography ranged from 14,000 to 18,500. The amino acid compositions of the beta-SCP and gamma-SCP from BSC and BSR were similar except that beta-SCP from BSR lacked half-cystine whereas gamma-SCP from BSR contained three times as much half-cystine as the SCP forms prepared from BSC. All forms of SCP showed reactions of identity when compared by immunodiffusion analyses with a rabbit anti-bovine SCP serum; none formed precipitin lines with a rabbit anti-bovine myelin basic protein (MyBP) serum.


Cell-mediated immunity to Friend virus-induced leukemia. II. Characteristics of primary cell-mediated cytotoxic response. The primary cell-mediated cytotoxic response to a Friend virus-induced leukemia, FBL-3, in C57BL/6 mice was measured by the 125IUdR release assay. Intraperitoneal (i.p.) inoculation of 1 x 10(1) FBL-3 cells produced progressive tumor growth (progressors); subcutaneous (s.c.) inoculation of as many as 5 x 10(6) FBL-3 cells produced only transient tumor growth (regressors), and these mice would subsequently resist i.p. challenge of FBL-3 cells at 3 days after s.c. inoculation. The kinetics of the primary cell-mediated cytotoxic response of regressors was biphasic. Significant cytotoxicity could be detected at 3 to 5 days after s.c. inoculation of 5 x 10(6) FBL-3 cells peaked at days 10 to 14, declined to a very low level or became undetectable around days 20 to 30; then the reactivity reappeared and persisted at least up to 60 days. In progressors, the kinetics of the cell-mediated cytotoxic response was similar to the regressors, but the reactivity was much lower. The cytotoxic response was found to be T cell dependent, during both the first peak (days 10 to 14) and the second peak (days 40 to 60). In adoptive transfer experiments, lymphocytes from regressors gave 90% protection against i.p. challenge of FBL-3; lymphocytes from progressors only gave 40% protection.


Cell-mediated immunity to Friend virus-induced leukemia. III. Characteristics of secondary cell-mediated cytotoxic response. By employing the 125IUdR release cytotoxicity assay, we have been able to measure the primary and secondary cell-mediated cytotoxic response of C57BL/6 mice to FBL-3 cells, a syngeneic Friend virus-induced leukemia. It was found that the secondary cell-mediated cytotoxic response occurred more rapidly after challenge (within 3 days) than the primary response, and the levels of reactivity were considerably higher. As in the primary response, the secondary cytotoxic reactivity of spleen cells was T cell dependent, being eliminated by pretreatment with anti-theta antibody plus complement. However, the secondary reactivity of pertioneal exudate (PE) cells was not entirely T-cell dependent. The specificity of the secondary cytotoxic response was analyzed by primary or secondary immunization with various tumor cells and by testing of cytotoxic lymphocytes against a variety of target cells. When spleen cells were used for testing, only tumor cells induced by Friend, Moloney, or Rauscher (FMR) leukemia viruses could produce secondary cell-mediated cytotoxic responses against FBL-3 cells. This correlated well with the specificity observed in the in vivo tumor transplantation protection studies. Similarly, spleen cells immune to FBL-3 had appreciable cytotoxicity against tumor cells induced by FMR viruses. The FBL-3 immune mice also gave significant protection against the challenge of FMR leukemias. When PE cells were used for testing, they gave higher levels of cytotoxicity against tumor cells induced by FMR viruses, but also gave less, but appreciable, cytotoxicity against non-FMR tumors. The latter reactivity might be related to the antigens induced by the murine endogenous type C viruses.


Antigenic determinants of adenovirus capsids. II. Homogeneity of hexons, and accessibility of their determinants, in the virion. We have tested the two principal theories which explain the previous finding that small amounts of type-specific antibody to the adenovirus hexon can neutralize infectivity, whereas even large amounts of cross-reactive antibody do not. a) It has been suggested that the type-specific determinants are especially prominent in the virion. We have therefore measured the capacity of whole virus to bind appropriate antibodies, using a sensitive radioimmunoprecipitation (RIP) system. In fact, virions bound type-specific and cross-reactive antibodies impartially. Moreover, they bound both much less effectively than did free hexon or disrupted virus, suggesting that many of each kind of determinant are inaccessible in virions. b) It has been suggested that the type-specific determinants are confined to those hexons located next to the pentons, and that they are the targets for neutralizing antibody. We have therefore studied the antigenicity of peripentonal and nonamer hexons isolated from virions, and found that each possessed both kinds of determinants. Furthermore, these were present in the same proportion as in hexons purified from the soluble antigens in infected cells ("free hexons"). We concluded that the mechanism of neutralization by antibody is complicated, and that the type-specific determinants exposed on the virion must play a crucial role.


Regulation of helper cell activity by specifically adsorbable T lymphocytes. Mice immunized with soluble proteins such as human serum albumin (HSA) or ovalbumin (OA) develop in their spleens antigen-specific T and B lymphocytes. These populations of lymphocytes can be separated from each other by different means; e.g. treatment with anti-theta-antiserum and complement removes selectively T lymphocytes, whereas passage through glass bead columns coated with mouse immunoglobulin (Ig): anti-Ig complexes creates a relatively pure population of T lymphocytes. During the course of such separation studies it was observed that the helper capacity of HSA (or OA) immune mouse spleen cells after Ig:anti-Ig column passage frequently was higher than expected from the enrichment in theta-positive cells. In addition, after adsorption onto antigen coated Bio-Gel beads this effect was even more pronounced, i.e., and increase in the relative helper capacity of about 3 or 4 times compared with an increase in the content of theta-positive cells from about 30% to 40 to 50% after adsorption. The present results will demonstrate that the increased helper capacity was a specific phenomenon which was regulated by theta-positive cells. The regulatory cells specifically adsorbed onto antigen-coated Bio-Gel beads have not been successfully eluted by EDTA or excess-free antigen so far, and they were still adsorbed after pre-incubation with anti-Ig antibodies under conditions where specific B lymphocyte adsorption was almost prevented.


An improved fluorescence probe cytotoxicity assay. The phenanthridine dye, ethidium bromide, which is actively excluded by viable cells, undergoes a significant fluorescence enhancement at 5900 A upon binding intracellular double-stranded polyribonucleotides. A rapid and sensitive assay of antibody mediated cytotoxicity to cells grown in vitro has been developed using this phenomenon. In this communication, we describe this fluorescence probe cytotoxicity assay and a sensitive electro-optical system designed to measure the fluorescence enhancement of ethidium bromide as it intercalates with intracellular polyribonucleotides. Basic characteristics of the fluorescence enhancement resulting from the interaction of ethidium bromide and non-viable cells are presented as well as examples of this assay as it has been used to study surface membrane neoantigens of cells tranformed by the oncogenic DNA virus, SV40.


A method for staining multiple chemotaxis filters. A device is described which holds chemotaxis filters on glass slides so that the cells on as many as 128 filters can be stained simultaneously. The procedure for using the batch stainer with Wright's stain is outlined.


Detection of antibody in virus--antibody complexes by immunoferritin labelling and subsequent negative staining. A technique for the immunoferritin labelling of antibody participating in virus--antibody complexes has been developed. Complexes were labelled by incubation with the immunoferritin reagent and excess reagent removed by polyacrylamide gel electrophoresis. Labelled complexes were then visualised in the electron microscope by negative staining.


Mixed tumors of the skin of the salivary gland type: a light and electron microscopic study. The light and electron microscopic features of two mixed tumors of the skin of the salivary gland type demonstrate eccrine differentiation of the epithelial component. The tumors were made up of tuboalveolar spaces lined by an eccrine duct type of epithelium with luminal cells showing numerous microvilli. The mesenchymal elements were fibroblasts and chondrocytes embedded in a mucous and chondroid matrix. Groups of undifferentiated cells without eccrine differentiation but with ultrastructural features suggestive of epithelial origin were seen among the mesenchymal and epithelial elements of the tumor.


Dibutyryl cyclic AMP-induced differentiation of epidermal cells in tissue culture. Histochemical and biochemical techniques have been used to compare the effects of dibutyryl cyclic AMP on epidermal cells and dermal cells in primary tissue culture. Rhodamin B staining showed only scattered positive cells in nontreated epidermal cells and a few contaminating keratinizing cell foci in both nontreated and treated dermal cell cultures. In contrast, treated epidermal cells stained strongly and had many keratinizing cell foci. A significant increase in histidine, cystine, and arginine incorporation was noted in epidermal cells treated with dibutyryl cyclic AMP as compared to untreated epidermal cells and to dermal cell cultures both treated and untreated. Dibutyryl cyclic AMP had no significant effect on leucine and phenylalanine incorporation. These results seem to suggest that the intracellular level of cyclic AMP not only controls the synthesis of DNA by epidermal cells in culture but also induces the process of differentiation toward keratinization.


A new hepatitis B virus surface antigen. A new determinant of hepatitis B surface antigen (HBs Ag), g, which is distinct from the previously identified determinants a, d, y, w, and r, was studied. This new antigen tended to be associated with the d antigen; it was found in 97% of ad specimens and in 15% of ay specimens. With few exceptions, the atypical subtypes, adg- and ayg+, were identified in sera from blood donors, but not in sera from patients with acute viral hepatitis; this finding possibly reflects a reduced tendency of these viral strains to evoke a host immune response. The ability to identify g represents a refinement of HBs Ag serotyping that may be of value in the study of the epidemiology of hepatitis B virus.


Effects of IgM and IgG antibody in patients with bacteremia due to gram-negative bacilli. Earlier studies, which indicated that high titers of O-specific antibody to the patient's infecting organism in acute-phase serum specimens were not associated with a decrease in the frequency of subsequent shock and death in bacteremia due to gram-negative bacilli, were reexamined for evaluation of the protective activity of specific IgG and IgM antibody. Titers of hemagglutination antibody and levels of IgM, determined by indirect immunofluorescent staining of the patient's infecting organism, as well as hemagglutination titers after reduction of serum with 2-mercaptoethanol and IgG levels, correlated closely (P less than 0.001). High titers of IgG antibody to the patient's infecting organism in acute-phase specimens were associated with a significant reduction in the frequency of shock and death in bacteremia. In contrast, high titers of IgG antibody were not associated with a diminution in the frequency of shock and death. The previously demonstrated protective activity of antibody to an antigen, Re lipopolysaccharide, shared by most gram-negative bacilli was reconfirmed and shown to be independent of the protective activity of O-specific IgG antibody.


Evaluation, surveillance and treatment of panoral leukoplakia. Leukoplakia is well-recognised as a premalignant condition of the oral mucosa. Despite many attempts to produce definitive laboratory tests for the prediction of those cases of premalignant leukoplakia which may undergo malignant transformation, there is as yet no fool-proof method of clinical and laboratory assessment. The problem is nowhere more manifest than in the management of panoral leukoplakia, especially in frail, elderly and apprehensive patients. In such cases the selection of sites for biopsy and treatment is often more empirical than logical. The introduction of cryosurgical techniques for the treatment of leukoplakia has enabled the surgeon to eradicate panoral white patches in a more conservative fashion. The application of a diagnostic nuclear staining test using toluidine blue dye, coupled with local exfoliative cytology, has provided a system for evaluation and surveillance of panoral leukoplakia which is clinically simple and at least as reliable as any predictive test previously described.


The effects of intra-arterial bleomycin therapy on squamous cell carcinoma of the oral cavity. Biopsy and autopsy examinations. The results of a clinico-pathologic study of 7 cases of advanced squamous cell carcinoma of the oral cavity are presented, which were treated by intra-arterial perfusion of bleomycin. In 5 cases, no histologically vital residual tumour could be found locally after therapy. The tumour area shows advanced fibrosis, and ulcerated areas have re-epithelized. The tumour cells are devitalized by keratinisation, while simple necrosis plays a minor role. Following the keratinisation there is a marked resorptive granulomatous inflammation with giant cells typical of a foreign body reaction and fibrous organisation. The special type of reaction of the tumour cells with keratinisation seems to be important for biopsy examinations and for therapeutic considerations. It explains the therapeutic success in highly differentiated squamous cell carcinomas with a tendency for keratinisation. For the first time a "cell-specific, cytocidal" therapy seems to be possible. Previous or simultaneous therapy with methods which might cause a de-differentiation of the tumour, seems to be harmful. Also with intra-arterial perfusion the lung must be considered as the target for the most serious side effects. In one case a bleomycin-induced lesion of the lung was the cause of death, in 4 cases a bronchopneumonia. Although in all cases the therapeutic chances and the prognosis were poor, 4 patients were cured of the tumour. One patient has lived without recurrence or metastases of the tumour for 20 months.


A teaching bank of audiovisual materials for family practice. Although increasing emphasis has been placed in recent years on the production and use of audiovisual materials in medical education, little work has yet been done on the identification and application of these materials in family practice teaching programs. This paper describes the content, uses, limitations, and initial experience of a Teaching Bank developed to support family practice teaching in varied settings. Video cassette and tape-slide units are most useful; audio cassettes alone are less likely to be selected. The evaluation of content, quality, and effectiveness of audiovisual media poses a particular problem. Although audiovisual materials can enhance learning based on different individual learning needs and styles, they cannot stand alone and usually must be supplemented by other teaching methods.


Immunological relationships of bacterial L-asparaginases. Rabbit antisera against L-asparaginase preparations from Escherichia coli, Erwinia carotovora, Citrobacter sp. and Chromobacterium violaceum showed on immunoelectrophoresis that only the enzymes from E. coli and Citrobacter are immunologically related. Purified preparations had to be used to determine the immunological cross-reactions. Immunoelectrophoresis at different pH values yielded the zero mobility points of the enzymes. The activity of the Er. carotovora preparation was enhanced up to fourfold by homologous antiserum but not by normal sera. Heterologous antisera also enhanced, but only at a higher concentration. Less enhancement was observed for the other enzymes with antisera as well as with bovine serum albumin. Inhibition was not observed.


Characteristics of the major internal protein and RNA-dependent DNA polymerase of bovine leukaemia virus. A virus designated bovine leukaemia virus (BLV), associated with leukaemia in cattle and previously demonstrated to induce the disease in sheep, was purified from chronically infected sheep cell cultures. Electrophoretic analysis showed a major protein of mol. wt. about 24,000 (p24) which reacted in gel diffusion and complement-fixation tests with sera from naturally infected cattle, experimentally infected sheep, and guinea pigs immunized with p24. BLV p24 has an isoelectric point of 8-6. Interspecies antigenic reactivities characteristic of mammalian Type C virus p30s were not detected in disrupted BLV or on p24. Sheep and guinea pig antisera to BLV, reactive with p24, also did not precipitate several Type C virus p30s in radioimmunoassays. BLV is also distinguished from Type C viruses and resembles mouse mammary tumour virus and Mason-Pfezer virus in having an RNA-dependent DNA polymerase which is preferentially active in the presence of Mg++ when synthetic templates are used. Along with previously published morphological data, the above indicates that BLV is not a Type C virus as classically defined. Four hundred and forty one human sera from cancer patients and matched controls were non-reactive with disruped BLV, BLV infected cells, and BLV p24 in complement-fixation tests.


Detection of simian type-C particles in Mason-Pfizer virus infected cultures. A simian type-C virus has been detected in cultures chronically infected with Mason-Pfizer monkey virus (M-PMV). Simultaneous budding of M-PMV and type-C virus particles from the same cells was observed in cultures incubated at 37 or 40 degrees C. However, the frequency of such cells was greater in cultures grown at 40 degrees C. Although clusters of type-C viral buds were seen at the surface of the cells, extracellular mature type-C particles in cell pellets or concentrated virus preparations were very rarely found. The increase in frequency of type-C buds was found to be transitory since cultures adapted to growing at the high temperature demonstrated budding type-C particles only occasionally. Cultures producing type-C buds were found to contain, in addition to M-PMV antigens, serological activity with polyvalent antisera produced against multiple structural components of endogenous baboon virus (BV) or simian sarcoma virus (SiSV). The reactivity, however, was found not to be serologically related to the major SiSV P28 core protein.


Comparison between the effect of colchicine and lumicolchicine on axonal transport in rat motor neurons. The mitotic inhibitor colchicine (COL) and its isomer lumi-colchicine (lumi-COL) were tested on the axonal transport of acetylcholine (ACh), cholineacetyltransferase (CAT) and ACh-esterase (AChE) in rat motor nerves in vivo. COL was found to be far more effective in blocking axonal transport of all 3 substances than was an equimolar concentration (0.1 M) of lumi-COL, when injected into the sciatic nerve. Since COL and lumi-COL have similar properties as regards binding to cellular membranes, but differ markedly in their binding capacity to microtubule protein, the results give strong support for the theory that microtubules are essential for intra-axonal transport in mammalian nerves.


Graphic symbols in an occupational health service. The use of graphics is newly entered in the communications arena and is limited only by the producer's imagination and the clarifying skills of the artist. We described the use of graphics in an occupational health setting, and the basic principles of their development. It is suggested that others using original graphics, or considering their adoption, deposit specimen copies with the Central Symbols Archives.


Alpha1-antitrypsin in amniotic fluid and cord blood of preterm infants with the respiratory distress syndrome. Total protein, alpha1-antitrypsin, and alpha2-macroglobulin were measured on amniotic fluid in 125 pregnancies between 11 and 42 weeks' gestation, and on the cord blood of 66 newborn infants. Amniotic fluid surface active material was assessed by the foam stability test. Amniotic fluid alpha1-antitrypsin is linearly and directly related to amniotic fluid total protein (r = 0.703, p less than 0.001). The cord alpha1-antitrypsin is also linearly related to cord total protein. Intrapartum complications are associated with a significant lowering of the cord alpha1-antitrypsin. Infants with a negative foam stability test had RDS regardless of the amniotic fluid alpha1-antitrypsin. The cord blood alpha1-antitrypsin value did not appear to be related to amniotic fluid surface active material. There were 23 infants with cord alpha1-antitrypsin of less than 0.2 gm/dl and with an intermediate or positive foam stability test; 19 of them had respiratory difficulties of varying severity. It is conceivable that infants, in spite of apparent adequate prenatal lung surfactant, develop respiratory disturbances on the basis of pulmonary fluid and protein transudation and/or reduction or inhibition of pulmonary surfactant incident to intrapartum complications.


Comparative pharmacokinetics of coumarin anticoagulants XV: relationship between pharmacokinetics of dicumarol and warfarin in rats. The distribution, elimination, and anticoagulant effect of dicumarol and warfarin were determined in adult males rats following intravenous injection of single doses of these drugs in crossover experiments. The biological half-life of dicumarol ranged from 5 to 28 hr; that of warfarin ranged from 9 to 30 hr. There was a statistically significant correlation between the following pharmacokinetic characteristics of dicumarol and warfarin in individual animals: biological half-life, apparent volume of distribution, total plasma clearance, and concentration in plasma eliciting one-half the maximum anticoagulant effect (effective concentration). The mean ratio of the respective biological half-lives (warfarin/dicumarol) was 1.42, and that of the apparent volumes of distribution was 1.50. The ratio of the effective plasma concentrations (dicumarol/warfarin) was correlated negatively with the half-life of dicumarol and positively with the ratio of the half-life values (warfarin/dicumarol) in individual animals. Additional studies with serum samples from other rats showed pronounced interindividual differences in the serum protein binding of both dicumarol and warfarin and a strong correlation between the protein binding of these two drugs in serum of individual animals. The results of this study, together with the results of previous studies in this series, indicate that serum protein binding is the major determinant of interindividual differences in the pharmacokinetics of dicumarol and warfarin in rats under these experimental conditions.


The influence of fasting and stress on the response of rats to warfarin. A study was undertaken to investigate the influence of fasting (24 hours), epinephrine (four 0.25 mg/kg s.c. doses at hourly intervals), adrenocorticotropin (two 40 I.U. s.c. doses at 2-hour intervals) and immobilization stress (4 hours) on the response of rats to some coumarin or indanedione anticoagulants. The anticoagulants were always administered first and were followed immediately or within the next hour by the appropriate challenge. Fasting produced a significant enhancement of the antiprothrombin response to warfarin (0.5 mg/kg i.v. or 0.75-3.0 mg/kg s.c.), bishydroxycoumarin (7.5 mg/kg i.p. or 10 mg/kg s.c.) and phenindione (40 mg/kg p.o). Epinephrine and immobilization stress, but not adrenocorticotropin, similarly prolonged the prothrombin time after warfarin (0.75-3.0 mg/kg s.c.). When used in the absence of anticoagulants, all challenges had no effect on the prothrombin time. In addition, fasting did not affect the response of anticoagulated animals to vitamin K. Plasma free fatty acids were significantly increased by the various challenges. The binding constant of warfarin to undiluted plasma proteins were decreased from the control value by a factor of 1.7 and 2.0 as a result of immobilization stress and fasting, respectively. Fasting per se increased the amount of bound endogenous free fatty acids per mole of protein; the latter parameter was further increased in the presence of warfarin. The present data show that fasting and stress enhance the anticoagulant response to warfarin and suggest that this might be due to an interference of endogenous free fatty acids with binding of warfarin to plasma proteins.


Dissociation between nerve-muscle transmission and nerve trophic effects on rat diaphragm using type D botulinum toxin. Small doses of botulinum toxin can produce partial blockage of transmitter release at the nerve--muscle junction. 2. Subthreshold e.p.p.s, 3--10 days after poisoning, show a distribution of amplitudes that is fitted by Poisson statistics. Successive e.p.p.s. in a short train show a marked facilitation. 3. Two weeks or more after poisoning with a dose of toxin that paralyses the whole muscle, when nerve--muscle transmission is in course of recovery, subthreshold e.p.p.s have an amplitude distribution that is fitted by binomial statistics. This property of transmission is similar to those described in newly formed nerve--muscle junctions, during embryogenesis or regeneration. 4. Muscle fibres with subthreshold transmission in the 5--10 day group of muscles were all supersensitive to ACh, as were a number of fibres in which nerve stimulation still produced an action potential. 5. Two weeks or more after poisoning, muscle fibres with subthreshold transmission had lost their extrajunctional ACh-sensitivity, as had many fibres with m.e.p.p.s of roughly normal frequency but no response to nerve stimulation. 6. In diaphragm muscles poisoned with botulinum toxin between 1 and 4 days previously, the rate of fast axonal transport of radioactively labelled proteins down the phrenic nerve is not greatly affected, but the amount of materials carried is reduced to about one quarter of normal. These labelled proteins accumulate in the intramuscular portion of the phrenic nerve, in or near the nerve terminals, to a much greater extent than in controls, showing that the normal release of some of these materials has been prevented by the toxin. 7. It is concluded that the blockage of the trophic effects of nerves by botulinum toxin is due to a blockage of release of trophic factors other than ACh. 8. The muscle nerve cannot maintain a muscle in its normal state simply by activation of contraction, and a regenerating nerve terminal can restore a muscle towards its normal state before it can release enough ACh to produce muscle contraction.


The application of long- and short-term progestagen treatments for oestrous cycle control in heifers. Progestagen-impregnated pessaries were inserted into 205 heifers on eight farms. Treatments were either pessaries only for 20 days or pessaries for 10 days combined with an intramuscular injection of 250 mg progesterone+7-5 mg oestradiol benzoate on the day of insertion. Pessary retention was low (86.7%) over the 20-day period, but high (93-6%) during the 10-day period. Calving rates of heifers inseminated in oestrus following the 20-day treatment were low, while those of heifers inseminated on a fixed-time basis with the 10-day treatment were slightly higher than those of control heifers.


Effect of long- and short-term intravaginal progestagen treatments on synchronization of oestrus and fertility in heifers. A total of 273 Herford cross heifers were treated with intravaginal progestagen pessaries to determine some of the factors affecting oestrous response and fertility following long-term (20-day) and short-term (10-day) treatments. Oestrous response and degree of synchronization were high after treatment for 20 days, but the fertility rate was lower than that of control heifers. There was no difference in the fertility of heifers inseminated artificially and those mated naturally. When the treatment period was reduced to 10 days and 900 mg progesterone and 5 mg oestradiol valerate given intramuscularly at the start, a high oestrous response and a low degree of synchronization resulted, but the conception rate was similar to that of the control animals. Reducing the dose of progesterone to 250 mg resulted in a high oestrous response and a high degree of synchronization. The stage of the cycle at the start of the 10-day treatment did not affect the oestrous response. Retention of the progesterone pessary was low (79-9%) in heifers treated for 2- days, but was 100% in those treated for 10 days.


Block of hamster fertilization by anti-ovary antibody. Antibody produced in rabbits against hamster ovary blocked fertilization of golden hamster eggs in vitro by binding to the surface of the zona pellucida and rendering it impenetrable to spermatozoa. The antibody was also effective in blocking fertilization in vivo. An intraperitoneal injection of the antibody into females resulted in the complete inhibition of fertilization for approximately 12 days, i.e. three oestrous cycles. This temporary sterility was apparently due to the binding of the antibody to the zona pellucida of oviducal and ovarian oocytes, and dot due to a failure of sperm ascent or capacitation in the female genital tract. Neither the oestrous cycle nor ovulation was affected by the antibody injection.


Stereochemical analogs of a muscarinic, ganglionic stimulant. 2. Cis and trans olefinic, epoxide, and cyclopropane analogs related to 4-[N-(3-chlorophenyl)carbamoyloxy]-2-butynyltrimethylammonium chloride (McN-A-343). Preparation of analogs of 4-[N-(3-chlorophenyl) carbamoyloxy]-2-butynyltrimethylammonium chloride [1 (McN-A-343)], cis- and trans-4-[N-(4-chlorophenyl)carbamoyloxy]-2-butenyltrimethylammonium iodides (5 and 6), and the corresponding epoxides and cyclopropanes is reported. Pharmacological testing for ganglion-stimulating activity demonstrated that the trans olefin 6 and trans epoxide 8 have properties similar to 1, while the trans cyclopropane analog 10 was inactive. All cis compounds were inactive. The muscarinic ganglion-stimulating properties of the active compounds are interpreted in terms of similar fit at the receptor level by the alkyltrimethylammonium ion and the ether oxygen 5.7 A distant, as well as an electron-rich center midway between groups in the form of a double bond or unshared electron pairs. Comparison of smooth muscle and ganglion-stimulating properties of the compounds showed that trans epoxide 8 was the most selective for muscarinic ganglionic sites.


Stereochemical analogs of a muscarinic, ganglionic stimulant. 3. 2,3-Substituted bicyclo(2.2.1)hept-5-enes and -heptanes related to 4-(N-(3-chlorophenyl)carbamoyloxy)-2-butynyltrimethylammonium chloride (McN-A-343). Preparation of analogs of 4-[N-(3-chlorophenyl)carbamoyloxy]-2-butynyltrimethylammonium chloride (1, McN-A-343), the isomeric 2-trimethylammoniomethyl-3-[N-(4-chlorophenyl)carbamoyloxymethyl]bicyclo [2.2.1]hept-5-ene iodides (10-13), and the corresponding -bicyclo[2.2.1]heptane iodides (14-17) are reported. None of the compounds demonstrated ganglion-stimulating activity similar to 1 or antagonized the effects of 1.


Partial characterisation of an inhibitor of streptolysin O produced by bacterial growth in serum. An inhibitor of streptolysin O is generated in human and animal sera by the growth of certain organisms. The ability to do this occurs most often in Pseudomonas aeruginosa and Staphylococcus aureus (in 90% and 86% of strains respectively), but in only 32% of Staph. epidermidis strains. The inhibitor is not formed in broth. The effect appears slowly on incubation, with maximum activity after 4-7 days. Evidence suggests that two enzymes are involved, an esterase which splits ester-bound cholesterol and a proteolytic enzyme which partially hydrolyses lipoprotein, resulting in cholesterol remaining attached to protein or polypeptide fractions but with some alteration of its spatial configuration such that it is now capable of attaching to streptolysin O. The inhibitory factor appears to prevent streptolysin becoming attached to cholesterol receptor sites on the erythrocyte membrane. Removal of the precursor from serum with magnesium carbonate suggests that low-density lipoproteins may be the precursor of the inhibitor.


Factors determining the susceptibility of mice to experimental phycomycosis. Various factors influencing the susceptibility of C3H mice to lethal infection by the phycomycete Absidia ramosa were examined. Mice 19-21 days old were exposed to graded doses of A. ramosa spores by various routes. After intravenous inoculation with doses in excess of 10(3) viable units, a variable proportion of mice developed lethal phycomycosis of the central nervous system within 2-8 days. The proportion of mice affected was related to the inoculum size, doses of 5 X 10(7) spores producing lethal infection in 90-100% of the mice. The disease was characterised by signs of involvement of the central nervous system appearing 8-12 h before death. At necropsy, fungal hyphae, frequently surrounded by infiltrations of mononuclear cells, could be demonstrated in the brain. Lesions were also often present in the kidneys; in other organs they were rare, but the presence of viable fungal spores could be detected by cultural procedures. Subcutaneous inoculation of A. ramosa spores did not produce lethal infection but intraperitoneal inoculation occasionally did so. Direct intracerebral inoculation invariably produced lethal infection even with very small doses of spores. The clinical and histopathological features of the disease were almost identical with those resulting from intravenous inoculation. Pre-inoculation treatment with azathioprine, cyclophosphamide and antithymocyte serum did not increase susceptibility. Susceptibility was increased by injections of zymosan, aggregared gamma-globulin, cortisone acetate and Fe(II) salts and by reticuloendothelial blockade. These treatments increased the proportion of mice developing lethal phycomycosis of the central nervous system, and in the case of cortisone acetate, also promoted disseminated infection. It was concluded that the natural resistance of mice to A. ramosa infection was probably dependent upon the activity of phagocytic cells, possibly acting in conjunction with complement and other non-specific serum factors.


Analysis of platelets from patients with thrombocythemia for reverse transcriptase and virus-like particles. A preliminary analysis of an RNA-directed DNA polymerase was made and a C-type virus-like particle was identified in platelets from 2 patients with the myeloproliferative disorder thrombocythemia (primary, essential, hemorrhagic, or idiopathic thrombocythemia). Platelet homogenates were centrifuged through a sucrose equilibrium density gradient. Both endogenous and exogenous DNA polymerase activity was found at a density of 1.19 g/ml. No activity was seen at comparable densities in control gradients. Electron micrographs of thin sections of these platelets revealed a particle with the morphologic characteristics of a C-type virus; however, the diameter of this particle was about 80 nm, slightly lower than that commonly found for C-type particles. Critical-point dried specimens, from the fractions of the sucrose gradient at which DNA polymerase activity was found, contained particles of the same size and morphology as those in the thin sections.


Factor supprossing alpha1 fetoprotein production. The injection of sera of adult mice or of a 20% extract of small and large intestine, liver, kidney, spleen, thymus, and lungs of adult mice suppressed the alpha1 fetoprotein production in syngeneic newborn recipients. Extracts of the striated muscles of adult mice had a meager inhibitory effect. Extracts of mouse embryonic intestine and those of the while embryo without the gastrointestinal tract had almost negligible amounts of suppressing factor. The possible role of humoral factors regulating the synthesis of embryonic antigens for the control of tumor growth was discussed author...


Assay in the mouse for delayed-type hypersensitivity to murine leukemia virus. Delayed-type hypersensitivity (DTH), assayed by footpad swelling, was induced in 6- to 8-week-old BALB/cCr mice immunized with formalin-inactivated, sucrose-banded murine type-C viruses. The DTH response was inducible with as little as 11.25 mug sensitizing antigen, was greatest after sc sensitization as compared to im and ip sensitization, and was optimally elicited with a 7-day challenge. A statistical evaluation of the DTH assay revealed that the test was consistently reproducible and limited only by biologic variability of the mouse and the standardization of the antigen preparation. The DTH response was specific for type-C virus subtypes because it could distinguish the Rauscher strain of murine leukemia virus from AKR leukemia virus when the challenge antigen was extracted with Tween 80-ether. Immunized mice that gave DTH responses were resistant to challenge with exogenous, live murine leukemia viruses.


An ether-sensitive antigen associated with bovine leukemia virus infection. An ether-sensitive antigen (es-Ag) associated with bovine leukemia virus infection was detected by immunodiffusion tests. This antigen was sensitive to ether, sodium periodate, and trypsin treatment. Based on column chromatography, es-Ag was a larger molecule than that of ether-resistant antigen (gs-Ag). With infection, precipitating antibody against es-Ag developed earlier than the precipitating antibody against the gs-Ag.


Cellular microcytotoxicity in human tumor systems: analysis of results. Using the tritiated-proline microcytotoxicity assay with cultured target cells, we tested a large series of melanoma, breast cancer, and bladder cancer patients for the presence of cell-mediated immunity. Specific, disease-related activity was infrequently observed, since the patients' lymphocytes exhibited selective activity against both disease-related and non-disease-related target cells. Most normal controls also demonstrated selective activity against these target cells. Neither the length of time the target cells had been cultured in vitro nor technical aspects of the assay, including the lymphocyte preparation methods, seemed to account for our results. We concluded that the experimental design of these tests may be the critical factor responsible for many of the disparate results being observed in different laboratories.


In vitro activation, infectivity, and production of endogenous type-C virus from OM rats. T24C, a continuous cell line derived from the pooled thymic tissue of normal inbred OM rats, spontaneously produced type-C virus. The virus genome was expressed cyclically. The amount of RNA-dependent DNA polymerase (RDP) and the number of 1.14 g dense particles/ml fluctuated simultaneously with cultivation. The released virus, RPT24C, did not infect cell lines from the rat, mouse, dog, or human. T31, also a rat thymus line, during its 2.5 years of cultivation did not produce type-C virus. Cocultivation with potentially permissive lines did not rescue any virus. 5-lodo-2'-deoxyuridine treatments at earlier passages yielded negative results. Chemical treatment at passages 111, 116, 123, and 128 yielded varying amounts of 3H-uridine incorporation at a sucrose density of 1.14 g/ml. Enzyme assays on chemically treated T31 cultures tested at passage 111 showed a small but transient burst of RDP activity. T31-B, a subline of T31, which was frozen and thawed once, released rat type-C virus spontaneously at passage 56. Two additional sublines of T31 (NI-T31 and NII-T31) were maintained for 2.5 years in culture without any cell-dispersing treatment. NI-T31, but not NII-T31, spontaneously released type-C virus. Once induced, the type-C viruses from T31-B and NI-T31 were continuously produced.


In vitro inhibition of cell-mediated cytotoxicity against syngeneic Friend virus-induced leukemia by immunoregulatory alpha globulin. Immunoregulatory alpha-globulin (IRA) derived from normal human plasma decreased cytotoxic reactivity as measured by an in vitro 5-iodo-2'-deoxyridine release assay of immune mouse lymphocytes against the syngeneic Friend virus-induced leukemia, FBL-3. This inhibitory effect depended on the dose of IRA used and was not due to the cytotoxic effects of IRA on the effector cells or target tumor cells. We also found elevated levels of serum alpha-gloubins in FBL-3 tumor-bearing mice as compared to normal mice. These data and the demonstration of decreased specific cytotoxic reactivity in FBL-3 tumor-bearing mice suggest that IRA functions in the suppression of the host's immune response against tumors.


Tracheobronchial and pulmonary cytologic changes in shock. Smears were made from tracheobronchial washings of patients undergoing general endotracheal anesthesia for surgery. Morphologically abnormal histiocytes were noticed in specimens obtained from subjects during hemorrhagic shock. These cells were overloaded with Papanicolaou stain (a greenish orange compound) which compressed the nucleus against the cell membrane. Cytochemical staining methods were undertaken to discover the composition of this substance. In secretions suctioned from ten patients in shock, large numbers of histiocytes were found to have ingested inorganic iron detectable by the Prussian blue method. Only two patients from a matched control group had smears in which this phenomenon was discovered. The maximum proportion of histiocytes containing Prussian blue granules was 40% for patients in shock and only 2% in normal controls. Histologic studies conducted on rats submitted to hemorrhagic shock were carried out to investigate this phenomenon. Iron-laden histiocytes were found in the kidneys, spleen, and lungs of both shocked and control animals. However, substantially more histiocytes containing Prussian blue positive granules were discovered in the lungs of rats in shock than in controls. It may, therefore, be that iron is deposited in the lungs in low flow states.


Synthesis of long, representative DNA copies of the murine RNA tumor virus genome. Virions of Moloney murine leukemia virus can synthesize two classes of DNA molecules complementary to their 70S RNA. One class consists of molecules about 200 nucleotides long, which are of limited sequence complexity; these molecules are formed preferentially if the dNTP concentration during the reaction is low. The second class consists of very heterogeneous DNA molecules with weight-average size of about 1,000 nucleotides containing at least 70% of the viral RNA sequences in approximately equal concentration. The longest of these molecules can be 5,000 nucleotides long. This second class of DNA is formed in large amounts only in reactions containing dNTP concentrations of 0.2 mM or higher. In such reactions after 24 h of incubation, at least 35% of the input RNA is represented in DNA copies. The ability to make long, representative DNA transcripts of tumor virus RNA provides a source of excellent probes for molecular hybridization.


Induction of type C virions from normal rat kidney cells by 2-deoxy-D-glucose. The sugar 2-deoxy-D-glucose (2-DG) induced the release of type C virions from an established line of normal rat kidney (NRK) cells. Within 20 h after the addition of 5 mg of 2-DG per ml to exponentially growing NRK clutures, more than 80% of the cells expressed the mammalian type C virus interspecies-specific antigen (p30) as determined by indirect cytoplasmic immunofluorescence. Maximal virion release occurred 1 to 2 days after 2-DG was added for 24 h to the growth medium although a low level of virion production was detected as early as 2.5 h after 2-DG treatment. Studies with inhibitors of RNA synthesis indicated a requirement for de novo RNA synthesis after the addition of 2-DG. Sensitivity of NRK cells to type C virion induction was limited to a relatively short period of in vitro growth and preceded spontaneous virion release by 8 to 10 subculture generations. A model is presented for the sequential derepression of latent type C virus information in serially propagated NRK cells.


In vitro transcription of 70S RNA by the RNA-directed DNA polymerase of Rouse sarcoma virus: lack of influence of RNase H. The influence of Rous sarcoma virus (RSV)-associated RNase H on the in vitro synthesis of DNA by the RSV RNA-directed DNA polymerase was determined under conditions whereby RNase H activity was selectively inhibited with NaF. Not only were we unable to detect any effect on the size, structure, or genetic complixity of the DNA product synthesized in the absence of RNase H activity, but the displacement of DNA from the 70S RNA:DNA hybrid structures was also unaffected. The suitability of 70S RNA:DNA hybrid structures synthesized in vitro to serve as a substrate for RNase H is discussed.


Species and interspecies radioimmunoassays for rat type C virus p30: interviral comparisons and assay of human tumor extracts. The major internal protein, p30, of rat type C virus (RaLV) was purified and utilized to establish intra- and interspecies radioimmunoassays. Three rat viruses were compared in homologous and heterologous intraspecies assays with no evidence of type specificity. The only heterologous viruses to give inhibition in these species assays were the feline (FeLV) and hamster (HaLV) type C viruses; these reactions were incomplete and required high virus concentrations. An interspecies assay using a goat antiserum prepared after sequentially immunizing with FeLV, RD 114, and woolly monkey virus p30's and labeled RaLV p30 was inhibited by all mammalian type C viruses, although preferentially by RaLV, FeLV, and HaLV. Thus, as in a previously reported assay developed with HaLV p30, rat, hamster, and cat p30's seem more closely related to each other than to mouse type C virus p30. High levels of specific antigen were found in all cell lines producing rat virus, whereas embryonic tissues from several rat strains and cell lines considered virus-free based on other tests were negative for p30. Rats bearing tumors containing Moloney murine sarcoma virus (RaLV) did not contain free circulating antibody to RaLV p30. Fifty-one human tumor extracts (including two tumor cell lines) were tested for activity in the RaLV species and 47 in the interspecies assays after Sephadex gel filtration and pooling of material in the 15,000- to 40,000-molecular-weight range. At a sensitivity level of 7 ng/ml (0.7 ng/assay) in the interspecies assay, all human tissues, with one exception, were negative. The one positive result is considered nonspecific based on proteolysis of the labeled antigen. Input tissue protein of the purified tumor extracts averaged 1.9 mg/ml with a range of less than 0.025 to 22 mg/ml. Tissues from NIH Swiss mice processed in the same manner were positive in the interspecies assay but negative in the intraspecies RaLV assay.


In vitro viral transformation of human prostatic carcinoma. Cell cultures derived from fetal human and prostatic adenocarcinoma were inoculated with SV40 virus. Within 6 weeks in vitro morphologic transformation occurred, after which the cells proliferated rapidly. They exhibited the SV40 T-antigen by immunofluorescence and survived propagation beyond the limited lifespan of uninfected cultures. We suggest that this may be a useful technique in establishing stable cell lines of human prostate.


Immunogenetic factors in preeclampsia and eclampsia. Erythrocyte, histocompatibility, and Y-dependent antigens. Determination of ABO and HL-A antigens and sex ratio of infants born to 46 women with preeclampsia or eclampsia, in comparison with normal controls, disclosed no predominant blood group, HL-A haplotype, or qualitative difference in maternal-fetal incompatibility. These results suggest that a link between immunologic mechanisms and the pathogenesis of toxemia syndromes, if present at all, must be associated with feto-placental tissue-specific antigens.


Role of preoperative administration of bleomycin and radiation in the treatment of esophageal cancer. The preoperative use of Bleomycin (BLM) alone or combined with irradiation in the treatment of esophageal cancer was evaluated in 58 patients who underwent surgical removal of the lesion for the past three and a half years. Histologic study showed such preoperative treatment is effective in inducing favorable changes in neoplastic cells in 69 per cent (40/58) of the primary lesions and in a significant number of involved lymph nodes of the second or the third groups. The study clearly suggests that preoperative Bleomycin administration combined with irradiation may be an useful adjunct preoperative procedure in esophageal cancer.


[Ventricular fibrillation in acute phase of myocardial infarct. 1. Relationship between the development of ventricular fibrillation in myocardial infarct and previous rhythm disorders]. The author analyses the results of experimental studies in dogs and cats, that included a continuous recording of ECG, electrogram and monophase cardiac potentials during 1 hour following coronary artery ligation. The ligation caused bradycardia, but no correlation was found between the degree of bradycardia and the development of extrasystole and ventricular fibrillation. Extrasystole developed in 100% of the experiments in which the coronary artery ligation resulted in ventricular fibrillation, and in 66% and 87% of those without this complication, conducted in dogs and cats respectively. The rate of extrasystole proved important for the prognosis of fibrillation. The number of extrasystoli noted during the mean time of the development of fibrillation was 5 times higher in the experiments with ventricular fibrillation than in those without fibrillation. In the experiments with fibrillation the extrasystoli tended to occur earlier within the cardiac cycle. Of the total number of extrasystoli, grouped extrasystoli comprised 89% in the experiments with ventricular fibrillation, and 21%--in those without fibrillation. Ventricular tachystystole was noted in 50% of the experiments with fibrillation and in 17% of those without this complication. In the experiments complicated by fibrillation the period of ventricular tachysystole was characterized by a gradual shortening of the cardiac cycles.


[Muscular neurotisation in traumatic abducens paresis (A new method of operative treatment) (author's transl)]. Traumatic abducens pareses occur quite often. The methods of operation known until now often achieved no good functional result. Therefore the authors experimentally tried to reinnervate by means of "musculr neurotisation" in 10 rabbits an extraocular muscle previously denervated. After 6 months the histological examination revealed the presence of newly germinated medullated nerve fibers and motor end plates. This was the proof that eye muscles may be reinnervated. Consequently in 5 patients with traumatic abducens paresis an implantation of the inferior oblique muscle was done. The functional improvement secured by this method of operation which for the first time has been applied on the human eye was good, in some cases even extremely good, depending on the time elapsed between lesion and operation.


Urinary proteins in Sprague-Dawley rats with chronic progressive nephrosis. By means of polyacrylamide concentration and electrophoresis on cellulose acetate strips, protein fractions have been determined in the urine of male Sprague-Dawley rats from 6 1/2-28 mo of age. An increasing percentage of albumin was expressed in the albumin/globulin ratios of 0.72, in a group of rats excreting a mean concentration of 1.91 mg/ml of protein in the urine, and 2.24 in a group of rats excreting, on the average, 17.62 mg/ml. A relative decrease in globulin, particularly of the alpha2 fraction, was shown. From a statistical model based on the regression parameters for 24-hr protein excretion and urine protein concentration, it was estimated that the doubling time for protein excretion was 3.2-3.69 mo. This interval is roughly one-half that reported for the Wistar rat.


Synthesis of adenosine triphosphate by way of potassium-sensitive phosphoenzyme of sodium, potassium adenosine triphosphatase. The sodium and potassium ion pump is an intrinsic enzyme of plasma membranes. In these experiments it was driven backward in a transient two-step operation involving, first, phosphorylation of the enzyme from inorganic phosphate, and second, transfer of the phosphate group from the enzyme to ADP upon addition of a high concentration of Na+. There was no evidence of a significant concentration gradient across the membranes. Na+ presumably reached the solutions on both faces of the membrane simultaneously and provided the energy for synthesis simply as a consequence of ligand binding. An interaction free energy between the free energy of the binding of Na+ and the free energy of hydrolysis of the phosphate group on the enzyme was estimated. The experiments also suggested a feature of the transport mechanism. This is control by phosphorylation of access pathways from the solutions in contact with the faces of the membrane to an active center for cation binding. In the dephosphoenzyme access would be to the intracellular solution and in the phosphoenzyme access would be to the extracellular solution.


Fixation of Celestin tube to the anterior abdominal wall. A new technique. Palliative intubation in the management of an unresectable carcinoma of the esophagus can be accomplished by "push through" or "pull through" techniques. The most frequently encountered complication is tube dislodgment-proximal or distal migration. Dislodgment occurs more freqeuntly with the "push through" tubes such as the Souttar tube than with the "pull through" tubes such as the Celestin tube. The incidence of migration reported for the "pull through" tubes varies from 3 to 14 per cent. In order to prevent migration, we sutured the tube to the anterior abdominal wall in 6 patients. The sixth patient developed a gastrocutaneous fistula and fasciitis at the point of anterior abdominal wall fixation. In the next 4 cases, we used a two-suture technique, in which no single suture passed from the gastric lumen to the anterior abdominal wall fascia. The Celestin tube was fixed to the gastric wall over a Dacron felt pledget and this pledget was then anchored to the rectus fascia. This technique has prevented both migration of the tube and complications related to gastric perforation.


Intracerebral haemorrhage. Accuracy of computerised transverse axial scanning in predicting the underlying aetiology. The diagnostic accuracy of computerised transverse axial tomography in predicting the pathology underlying cases of nontraumatic intracerebral haematoma has been reviewed. A 160 X 160 matrix was used and the scans of one hundred patients whose diagnoses had been confirmed by either angiographic or post-mortem examination were studied. In the absence of any clinical information a diagnosis of either cerebral aneurysm or primary intracerebral haemorrhage could be made with an accuracy of 90%. The haematomas most likely to be misinterpreted are those associated with angiomas and tumours, and these two lesions were responsible for six out of the ten incorrect diagnoses. In predicting the site of a ruptured aneurysm accuracy ranged from 100% for those involving the anterior cerebral artery complex to 66% for those few haematomas associated with aneurysms of the internal carotid/proximal middle cerebral artery aneurysm complex.


Pharyngeal lecithin/sphingomyelin ratios in newborn infants. Lecithin/sphingomyelin ratios were measured on hypopharyngeal secretions from newborn infants within 6 h of birth. L/S ratios of 1.8 or less correlated with the development of the idiopathic respiratory-distress syndrome. Low L/S ratios were more common in the shorter gestation infants, and there was a predominance of L/S ratios of more than 1.8 above 32 weeks' gestation. In multiple births, the first-born of triplets and quadruplets had higher L/S ratios than those subsequently born. This may indicate a link between fetal lung maturation and the onset of labour.


Prostaglandin E and F concentrations in human endometrium after insertion of intrauterine contraceptive device. The prostaglandin (P.G.) content of endometrium from fourteen women was examined before and 1-5 months after insertion of an intrauterine contraceptive device. A statistically significant increase in P.G.E but not P.G.F was observed. The P.G.E concentration increased in eleven to fourteen cases and fell in three. The P.G.F content increased in five cases and fell in eight.


Apparatus for patient-controlled administration of intravenous narcotics during labour. The intermittent intravenous administration of narcotic analgesics had advantages over intramuscular administration, but is often impracticable. The design of a prototype apparatus is described, from which analgesic, pethidine, can be self-administered intravenously during labour. The apparatus consists of a syringe pump and control system from which the patient can demand an increment of analgesic. The minimum interval between increments and the dose of the increment are preset. A demand for analgesia is only successful when the patient passes a reaction-time test, subsequently modified. The apparatus has been used by forty-two mothers in labour. The mean total dose self-administered (2.2 mg/kg) was well within the prescribed limits, suggesting that mothers regulated their demand. The apparatus appears acceptable and safe in labour. The apparatus will probably be of value in comparative studies of analgesics.


Virus-like particles demonstrated by freeze-squeeze technique in acute-phase serum of patients with HBAg-negative hepatitis. Electromicroscopy of fluid, squeezed from excised and frozen precipitation lines obtained by immunoelectrophoresis in agarose gels of acute-phase sera from patients with HBAg-negative hepatitis, revealed particles of 22, 27, and 31-41 nm. These precipitates and particles were not present in normal human sera and disappeared from the sera of the patients after recovery. These particles were still demonstrable after two years' storage at -20 degrees C.


Gentamicin resistance in Staphylococcus aureus. A clinical isolate of Staphylococcus aureus was resistant to gentamicin, kanamycin, and tobramycin by virtue of the production of a drug-inactivating enzyme. Attempts to establish the cellular location of the genetic determinants for enzyme synthesis have given equivocal results.


Acute bacterial meningitis in childhood. Incidence and mortality in a defined population. Cases of acute bacterial (excluding tuberculous) meningitis and meningococcal infection, occurring in children under 10 years of age in the North-West Metropolitan region between 1969 and 1973 have been studied retrospectively by case-note review. The risk of having an acute meningococcal infection was estimated to be 1 in 1090, and that of having an attack of haemophilus meningitis to be 1 in 1500, but the age of 10 years. The case-fatality rates for meningococcal infection, haemophilus meningitis, and pneumococcal meningitis were 10.9%, 5.7%, and 16.4% respectively. Meningitis was not diagnosed in life in 26 of 94 fatal cases (28%).


Melioidosis antibodies in Commonwealth soldiers. Titres of melioidosis haemagglutinating antibodies of 1/40 or more were found in 18 of 905 British, Australian, and New Zealand soldiers serving in West Malaysia. Previous mild unsuspected melioidosis seemed to be responsible for these positive titres, which were more common in men exposed to surface water at work and during recreation. This accords with the current view that soil and surface water is the normal habitat of Pseudomonas pseudomallei, the causal organism. Pyrexia of unknown origin after arriving in Malaysia was significantly more common in men with titres of 1/40 or more than in the remainder. It is suggested that mild melioidosis may present as pyrexia of unknown origin. Pyrexias of unknown origin should be investigated vigorously in patients who are in or who have visited endemic areas.


Ultrasound and human-placental-lactogen screening early detection of twin pregnancies. In the past five years gradual introduction of ultrasonic screening of pregnant women increased the ante-partum detection of multiple pregnancy from 60% in 1971 to 95% in the first half of 1975. At the same time the average gestational age for dection of multiple pregnancies diminished from thirty-three to twenty-five weeks. The corresponding figures for 1963 were 32% and thirty-six weeks. Plasma human-placental-lactogen concentrations were assessed for their value in selecting a smaller target group for subsequent ultrasonic screening. All but 2 of 39 twin pregnancies examined by single H.P.L. determinations had H.P.L. values more than 1 S.D. above the mean of the normal distribution. The use of plasma-H.P.L. screening might lower the proportion of patients requiring ultrasonography for antepartum diagnosis of multiple pregnancies to 16% of the total pregnant population.


Intensive antenatal plasmapheresis in severe rhesus isoimmunisation. Intensive antenatal plasmapheresis can significantly lower Rh-antibody levels in the blood of mothers carrying babies severely affected with rhesus haemolytic disease. Twenty-seven out of forty-four mothers were successfully delivered of babies that survived, after being treated with a combination of plasmapheresis and intrauterine transfusions. A further 52 mothers were treated by plasmapheresis along. When severe disease developed as an early stage in pregnancy the results were rather poor, but excellent results were obtained when severe disease developed relatively late in pregnancy. No serious complications were encountered in the mothers or babies.


Familial male breast cancer.

Familial male breast cancer. Infiltrating ductal carcinoma of the breast occurred in a total of six men from two families. In one family specimens from three men who had prophylactic mastectomies revealed focal intraductal hyperplasia, suggesting a familial tendency toward proliferation of mammary-duct epithelium. In the other family, benign and malignant breast lesions also developed in several women. Preliminary data suggest elevated urinary oestrogen excretion in three men from these families, implicating a defect in oestrogen production or metabolism in the pathogenesis of male breast neoplasms.


Oral disopyramide in prophylaxis of arrhythmias following myocardial infarction. Oral disopyramide given prophylactically following myocardial infarction has been compared with placebo in a double-blind trial using continuous-tape monoriting of the electocardiogram. It caused a significant reduction in the incidence of ventricular arrhythmias and of the various degrees of heart-block. There was a significant reduction of reinfarction during hospital stay in patients who had received disopyramide. Disopyramide appears to be a safe and effective oral therapy in the prevention of potentially serious arrhythmias following myocardial infarction.


Effect of low-dose heparin prophylaxis on arterial oxygen tension after high laparotomy. The effect of low doses of heparin (5000 units of sodium heparin every 12 hours for 5 days) on arterial oxygenation was studied in 24 patients in the postoperative period after upper abdominal surgery. Another 24 patients served as a control group. The arterial oxygen tension was the same in both groups preoperatively and was equally significantly reduced during the 1st postoperative day. During the 2nd day, oxygen tension rose in the heparin-treated group to values which no longer differed significantly from the peroperative level. In the control group the significant reduction persisted until the 4th postoperative day. The arterial carbon-dioxide tension did not differ between the groups, neither did it vary significantly between days. There were no clinical signs of large pulmonary embolism during the postoperative period, chest X-ray was normal in all patients examined and a photoscan was normal in 23 of 24 subjects studied. Low-dose heparin treatment may apparently shorten the period of postoperative hypoxaemia, probably by counteracting both large pulmonary emboli and microthromboembolism.


Soluble fibrinogen/fibrin complexes in pre-eclampsia. Significantly increased concentrations of soluble fibrinogen/fibrin complexes were found in plasma samples from ten normal pregnant women when compared with ten non-pregnant age-matched controls. In ten women with pre-eclampsia mean soluble complex concentration was more than three times that in the age, parity, and gestation matched pregnant controls. Soluble fibrinogen/fibrin complexes are also found in the plasma of patients in various hypercoagulable and thrombotic states, including disseminated intravascular coagulation. These findings provide additional evidence that pre-eclampsia is associated with disseminated intravascular coagulation.


Acute lethal carditis caused by high-dose combination chemotherapy. A unique clinical and pathological entity. An acute lethal myopericarditis has been observed in four out of fifteen patients receiving high-dose combination chemotherapy which includes cyclophosphamide 45 mg/kg/day for four days. In all cases the myopericarditis occurred 5-9 days after the initiation of chemotherapy, with dyspnoea, tachycardia, orthostatic hypotension, fluid retention, decreased voltage on electrocardiography, and pericardial effusion documented by echocardiogram, and progressed in 2 to 6 days to a fatal low-output state despite vigorous treatment. In three of the four patients, necropsy was permitted and revealed the unique pathological finding of fibrin microthrombi in capillaries, fibrin strands in the interstitium, and fibrin strands within the heart-muscle cells.


A simplified assessment of response to parathyroid hormone in hypoparathyroid patients. Hightly purified bovine parathyroid hormone (B.P.T.H) was given by injection and/or infusion to six normal volunteers and to patients with surgical hypoparathyroidism (five cases), idiopathic hyparathyroidism (five cases), or poeudo-hypoparathyroidism (six cases). Infusion and injection of B.P.T.H. produced very similar patterns of response in plasma adenosine 3' 5' cyclic monophosphate (cyclic A.M.P.) In all six normal volunteers and in the patients with surgical (five cases) or idiopathic (four cases) hypoparathyroidism who had injections of B.P.T.H., plasma-cA.M.P. had risen significantly within 5 min and the peak response was genereally observed 10 min after injection of hormone. In the five pseudohypoparathyroid patients who received injections of B.P.T.H., plasma-c?A.M.P. concentration increased only slightly or not at all after the hormone was administered. Unlike the traditional test for the investigation of hypocalcaemia, the test described here does not require collections of urine samples.


Antibodies to herpes-simplex virus in "normal" cerebrospinal fluid. Thirty samples of cerebrospinal fluid (C.S.F.) obtained during routine myelography were examined for antibodies against herpes-simplex virus (H.S.V.) by antibody-mediated cell-dependent immune lysis. Antiviral antibody was found in the C.S.F. of all ten patients who were subject to recurrent cold sores. Thirteen of the twenty patients who denied or could not recall any previous herpetic infection had a serum antibody, and nine of them also had C.S.F antibody. All patients with C.S.F. antibody also had serum antibody. These results support the suggestion the H.S.V. antibodies may be responsible for maintaining the latency of H.S.V.


Acute diabetic abdomen in childhood.

Acute diabetic abdomen in childhood. Three children presented as acute surgical emergencies due to undiagnosed diabetes mellitus. Where diabetic ketoacidosis mimicks the acute abdomen three clinical features are important in reaching the right diagnosis-namely, a history of polydipsia, polyuria, and anorexia preceding the abdominal pain, the deep sighing and rapid respirations, and severe dehydration.


Treatment of stress-induced upper gastrointestinal/hemorrhage with metiamide. The H-2-blocking antihistamine metiamide was used to treat 14 episodes of bleeding from the stomach or duodenum in eleven patients. In 11 instances bleeding was due to erosive gastritis or duodenitis and bleeding promptly ceased after one or two doses of 300 mg at 6 h intervals and did not recur as long as the drug was continued. In the 2 instances in which bleeding continued, chronic ulcers had eroded into major blood-vessels. There were no complications from the drug even in five patients with severe bone-marrow suppression after renal transplantation. Metiamide seems to be a safe and highly effective agent in the control of bleeding due to erosive gastritis.


Does phenylethylamine cause schizophrenia? The model psychosis associated with amphetamine overdosage is known to bear a close resemblance to acute paranoid schizophrenia. Amphetamine is chemically similar to the endogenous sympathomimetic amine, phenylethylamine, which possess many of its pharmacological properties. It is suggested that some cases of schizophrenia may be associated with an abnormal phenylethylamine response, either from increased concentrations of the amine or from abnormal receptor sensitivity to it.


Pregnancy: Nature's experimental system. Transient manifestation of immunological diseases in the child. Major advances in knowledge of immunological diseases have resulted from observation of transient effects on children borne by women with such diseases. The discoveries that in Graves' disease and myasthenia gravis there are IgG antibodies directed against receptors sites are examples of such developments, while "ikiopathic" thrombocytopenic purpura is now accepted as immunological owing to its behaviour during pregnancy. In some instances observations of transient neonatal forms do not correspond with the disease manifestations in the mother. These discrepancies may be due to surgical removal of an organ vital to the disease process; inactivating damage by the disease to such an organ; presence of a blocking antibody of a molecular type not transferred across the placenta; differing tissue-antigen specificity or differing lymphocyte cooperation based on genetic variation. At present there are unexplained observations of fetal/neonatal effects in relation to diabetes mellitus and systemic lupus erythematosus which suggest that study of immunological parameters might be profitable. Determination of the HLA status of mother/fetus pairs may give rewarding clues. In the elucidation of the diseases now proven as antibody-mediated in the antibodies first discovered often turned out to be irrelevant red herrings.


Long-stay patients with long-stay drugs. A case review; a cause for concern. The psychotropic medication of 200 long-stay psychiatric patients was reviewed by the consutant in charge of the ward, the ward doctor, and the ward sister. Those patients in whom medication was altered or stopped were reassessed by the same team after 6 months. About half the patients on medication had been receiving unnecessary or excessive medication.


Influenza at Christ's Hospital: March, 1974. Boys in a boarding-school given inactivated influenza-A or influenza-B vaccine have been observed during a simultaneous outbreak of influenza due to A/Port Chalmers, B/Hong Kong, and B/Intermediate strains. Influenza-B vaccine conferred substantial protection, the attack-rates in boys given B vaccine being 24% compared with 45% in a control group. A/Hong Kong vaccine, by preventing infection during the previous influenza A/England outbreak, left the boys vulnerable to influenza A/Port Chalmers. Influenza-A haemagglutination-inhibiting (H.I.) antibody acquired by natural infection was associated with more protection than H.I. antibody induced by vaccination. This difference was not seen with influenza-B H.I. antibody.


Mycoplasma T strains and perinatal death. In 23 out of 290 perinatal deaths in Addis Ababa, mycoplasma T strains were the only organisms responsible for congenital pneumonia and death. All but 2 of the infants were stillborn, 16 dying during the last six weeks of gestation. Infections apparently occurred through intact fetal membranes.


Specific B-cell antigens associated with gluten-sensitive enteropathy and dermatitis herpetiformis. Gluten-sensitive enteropathy (G.S.E.) and dermatitis herpetiformis (D.H.) are associated with an increased frequency of the histocompatability antigen HLA-B8. These diseases were found to be even more strongly associated with a specific B-lymphocyte surface antigen recognised by maternal antisera. Two antisera (B-1 and W-1) reacted with B lymphocytes form G.S.E. and D.H. patients. Antiserum B-1 reacted with cells from thirteen of sixteen G.S.E. patients and from fifteen of nineteen D.H. patients, while antiserum W-1 reacted with cells from fifteen of sixteen G.S.E. patients and from fifteen of fifteen D.H. patients. None of the sera tested reacted with B lymphocytes from thirty-seven normal individuals, whether or not they were HL8-B8 positive. The identification of this specific antigen provides further insight into the pathogenesis of G.S.E. and D.H. and might form the basic of a diagnostic test for these diseases.


Controlled trial of cysteamine in treatment of acute paracetamol (acetaminophen) poisoning. A randomised controlled trial of the use of intravenous cysteamine in the treatment of severe paracetamol poisoning has been performed. Thirty-eight patients presenting 3-17 h after ingestion were admitted to the trial; of these eighteen received cysteamine. Two patients died from hepatic failure, one in each treatment group. Analysis of the series as a whole showed no advantage of cysteamine in preventing biochemical abnormalities of liver function except for aspartate aminotranferase and serum ferritin levels, which were significantly less after cysteamine therapy. Separate analysis of the patients treated within 9 h of paractamol ingestion and of those treated 9-17 h after paracetamol ingesion similarly showed no definite advantage of cysteamine. Histological evidence of liver damage showed a possible beneficial effect of cysteamine. Cysteamine therapy did not prevent renal or pancreatic damage.


Atopic disorders and adult coeliac disease. A history of asthma, hay fever, and flexural eczema was significantly more common in patients with adult coeliac disease (A.C.D.) than in normal controls. Autoantibodies were also more common in A.C.D. First-degree relatives of A.C.D. patients were more likely than controls to have atopic disorders. A deficiency of local mucosal immunity due to abnormal IgA responses may underly this association between A.C.D. and atopy.


Hyaline-membrane disease, alkaline buffer treatment, and cerebral intraventricular halphaemorrhage. In 55 preterm infants dying from hyaline-membrane disease (H.M.D.) in 1971-74, infants with associated cerebral intraventricular haemorrhage (H.M.D./I.V.H.) had been given more intravascular sodium-bicarbonate solution, but the same proportions of cases with H.M.D. and H.M.D./I.V.H. received bicarbonate at the time of birth. Much of the sodium-bicarbonate solution given to H.M.D./I.V.H. infants was injected in response to the clinical effects of I.V.H. Maximum serum-sodium concentrations correlated with sodium-bicarbonate dosage but not I.V.H. The incidence of I.V.H. in preterm infants in 1971-74 was unchanged from 1956-59 when alkaline buffer treatment was not used. These findings do not suggest that sodium-bicarbonate therapy plays a major part in the pathogenesis of I.V.H.


Breath hydrogen in hyposucrasia.

Breath hydrogen in hyposucrasia. A simple and reliable test for the diagnosis of hyposucrasia is required, since this may be an unsuspected cause of long-standing gastrointestinal disorder. Furthermore little has been done to define the epidemiology of this condition, possibly because of the limitations of multiple blood-sampling. Breath hydrogen (H2) production after lactose ingestion is a reliable test for hypolactasia, and has now been measured after sucrose ingestion in eleven patients with various gastrointestinal symptoms. Six who had normal sucrase activity on jejunal biopsy produced no H2 after taking 50 g of sucrose. No H2 was produced in three patients with borderline hyposucrasia, either after 50 g sucrose or when retested using 100 g sucrose (two patients). However, the two patients with low jejunal sucrase activity showed rises of breath H2, after only 25 g glucose. Breath H2 measurement is a simple, accurate, and non-invasive test for diagnosing gastrointestinal symptoms due to hyposucrasia.


Magnesium and potassium content of normal heart muscle in areas of hard and soft water. Concentrations of both magnesium and potassium were significantly increased in normal heart muscle samples from an area with a soft water supply compared with matched samples from an area with hard water. The zinc contrentration was also slightly increased in the soft water area. The magnesium/potassium ratio was significantly lower in the softwater area. This relative deficiency of magnesium may increase the risk of death after myocardial infarction in soft-water areas.


Use of an elemental diet for long-term nutritional support in Crohn's disease. An elemental diet was used for long-term (up to 360 days) nutritional support in eight patients with Crohn's diesease. They had lost 30% (range 18-37%) of their weight in health. A whole-body monitor was used to measure total body potassium as an index of the lean tissue mass. Three preoperative patients who has been unable to maintain their weight on a normal diet were able to restore 10% per month of their lost lean tissue once they were established on an elemental diet. In five patients who had had a successful resection of their bowel lesion the postoperative rate of restoration of the lean tissue mass while taking an elemental diet was 18.5% per month, while in two control postoperative patients on a normal diet it was 19% per month.


Pathogenesis of enteritis necroticans in Papula New Guinea. It is suggested that enteritis necroticans occurs in the people of the Highlands of Papua New Guinea because they have low levels of digestive proteases in the intestinal lumen. These low protease levels are the result of a low-protein diet and the presence of heat-stable trypsin inhibitors in sweet potatato, the dietary staple. Low proteolytic activity allows the B toxin of Clostridium welchii type C, which is extremely susceptible to proteolysis, to initiate the disease.


Basic science, science, and medical education. It is generally accepted that medical students should learn those stories from the basic sciences which enable medicine to be rational. But there are several reasons why science as an explanatory activity has much more to offer, particularly if the views of Popper are accepted. Unfortunately an appreciation of the nature of science is only accidentally acquired in traditional schools and current forces may well weaken its chances in newer schools. Some of the criticisms levelled at modern medincine which may be attributed to excessive science are misdirected. There is too little science, not too much.


Mortality from lung cancer and coronary heart-disease in relation to changes in smoking habits. Changes in the type and quantity of cigarettes smoked in the United Kingdom from 1956 to 1971 are compared with changes in the dealth-rates due to lung cancer and coronary heart-disease (C.H.D.) from 1956 to 1973. Associated with a change in filter cigarettes there has been a decrease in lung-cancer mortality among men aged less than sixty years despite little change in the number of cigarettes smoked. In contrast, lung-cancer mortality has increased in women along with their cigarette consumption. C.H.D. mortality has continued to increase in both sexes, but to a greater extent in women. These changes are consistent with the hypothesis that, in tobacco smoke, tar is the principal aetiological factor in lung cancer, whereas carbon monoxide or other gaseous constituents are involved in the development of C.H.D.


Acute eye disease secondary to contact-lens wear. In a three-month survey of some hospital consultant ophthalmic services, 82 instances of complications of contact-lens wearing were reported. Some of the complications gave rise to discomfort, but in only 5 cases was vision impaired.