Neutrophil CD11b expression as a diagnostic marker for early-onset neonatal infection.

OBJECTIVES: To determine whether neutrophil surface expression of CD11b predicts early-onset infection or suspected infection in at-risk infants. STUDY DESIGN: CD11b expression on peripheral blood neutrophils was determined by flow cytometry of whole blood samples. Blood (0.1 ml) was obtained from a convenience sample of at-risk infants admitted to the neonatal intensive care unit, stained with antibodies detecting CD11b and CD15, chilled, and analyzed within 8 hours. Blood for culture, blood counts, and C-reactive protein (CRP) determination was obtained simultaneously. Subjects were grouped on the basis of culture results and clinical signs, and investigators were blinded to CD11b level. RESULTS: Of 106 subjects, seven had positive bacterial or viral cultures ("confirmed infection"), 17 had clinical signs of infection but negative cultures ("suspected infection"), and 82 had negative cultures and no clinical signs ("no infection"). Neutrophil CD11b was elevated in all infants with confirmed infection, 94% with suspected infection, and none with no infection. The negative and positive predictive values, sensitivity, and specificity were 100%, 99%, 96%, and 100%, respectively, for diagnosis of neonatal infection at initial evaluation. CD11b levels correlated with peak CRP (r2 = 0.76, p < 0.0001); however, CD11b was elevated at the time of admission in all five infants with proven bacterial infection, whereas CRP was normal until the second day in the neonatal intensive care unit in three of these five. Both infants with positive viral cultures had elevated CD11b, but the CRP levels remained within normal limits. The negative predictive value of neutrophil CD11b for identifying suspected or confirmed infection was 99%. CONCLUSION: This assay for neutrophil CD11b is a promising test for exclusion of early-onset neonatal infection. If validated prospectively, this assay may reduce hospital and antibiotic use in the population of neonates at risk for early-onset infection.

[Predictive experimental animal test of contact allergenicity. Relevance of the methods of the OECD and EEC guidelines. The organization for Economic Co-operation and Development]. / Prädiktive tierexperimentelle Kontaktallergenitätsprüfung. Relevanz der Methoden der OECD- und EG-Richtlinien.

In the first part of this article a historical note is given on the development of predictive contact allergenicity tests in guinea pigs starting with the Draize test, including epidermal methods and at least induction procedures using Freund complete adjuvant as a nonspecific enhancer of immunoreactivity. The tests of the OECD and EEC guidelines are listed and classified according to their predictive sensitivity. It is recommended to test reference allergens by different test methods in one's own laboratory to obtain personal experience with the various methods. Comparisons of the different methods based only on the literature are difficult, because there are so many influences on the performance of sensitization tests and because one is often not sure how the animals were termed positive, i.e. the threshold of a positive erythema score is often not clearly defined. Correlations between test results obtained in guinea pigs and man are presented for the Draize test, the maximization and the optimization test. It is emphasized that guinea pig tests are performed to assess the contact allergenic potential of a compound or formulation, but that the risk assessment is dependent on many other factors and should, therefore, be evaluated separately.

Predictive contact allergenicity influence of the animal strain used.

In order to determine the validity of reported alleged differences in the susceptibility of various strains of guinea pigs to the sensitizing action of known allergens, three unrelated strains of guinea pigs, namely Pirbright White, Hartley and Himalayan spotted strain, were selected. In our investigation, 3 haptens differing in their sensitization potential were used. The results showed that the sensitivity of the 3 given guinea pig strains was comparable. Furthermore, the sensitivity of a given guinea pig strain can only be determined by comparison of the sensitization reaction of several known contact allergens of various sensitizing potential.

[Comparative clinical trial of a new trihalogenated dermatocorticoid (halometasone) versus betamethasone dipropionate]. / Klinische Vergleichsprüfung zwischen einem neuen trihalogenierten Dermatokortikoid (Halometason) und Betamethasondipropionat.

In a multicenter controlled study carried out in Austria and Switzerland by 8 dermatologists in 208 patients with acute eczematous dermatoses, 0.05% halometasone cream proved to have significantly superior clinical efficacy than 0.05% betamethasone dipropionate cream (P much less than 0.05) showing an equal tolerability. Good to very good results were reported in 93% of the patients treated with halometasone and in 84% of those treated with betamethasone dipropionate creams (Diprosone, Diproderm). General tolerability of both treatments was good. Neither skin atrophy nor any systemic adverse effect, due to the transcutaneous absorption of the corticoids, were observed in this study.

Evaluation of halometasone ointment in the treatment of paediatric patients with chronic eczematous dermatoses.

Fifty children suffering from non-infected chronic eczematous dermatoses were treated with an ointment containing 0.05% halometasone, a new high potency trihalogenated synthetic corticosteroid. Halometasone ointment does not contain parabens or perfumes. The trial population consisted of patients (under 1 to 16 years old) with non-infected chronic nummular dermatitis, seborrhoeic dermatitis, atopic dermatitis and contact dermatitis. Halometasone ointment yielded very satisfactory therapeutic results and exhibited an excellent tolerability. It produced 'good' or 'very good' results in 82% of the patients and gave an overall cure rate of 76%. The number of patients obtaining an early cure, i.e. in less than 30 days, was thirty-six (72%). An early onset of therapeutic effect, i.e. within 6 days of starting the treatment, was reported in 77.1% of the patients. All the children (including eighteen infants) tolerated halometasone ointment well and no adverse effects, either due to local skin intolerability or the transcutaneous systemic absorption of the corticoid, were observed in this trial, nor were any instances of skin atrophy reported. Cosmetic acceptability and ease of application were considered 'good' in forty-nine (98%) patients.

Keratin-like proteins in normal and neoplastic cells of human and rat mammary gland as revealed by immunofluorescence microscopy.

Normal and neoplastic human breast tissue as well as lactating and nonlactating rat mammary glands and 7,12-dimethylbenz(alpha)-anthracene-induced mammary adenocarcinomas of rat, were examined by indirect immunofluorescence microscopy using guinea pig antibodies to human and bovine epidermal prekeratin and to cytokeratin polypeptide D from mouse hepatocytes. In normal mammary glands of both species, lactating rats included, the antibodies raised against human and bovine epidermal prekeratins strongly stained ductal and myoepithelial cells, whereas antibodies to hepatic cytokeratin D revealed, in addition, fibrillar staining in cells of the alveolus-like terminal lobular units and in milk secreting cells of the rat. The presence of some finely dispersed intermediate-sized filaments of the cytokeratin type in lactating alveolar cells of rat mammary gland was also demonstrated by electron microscopy. In human intraductal mammary carcinomas the antibodies to epidermal prekeratins showed staining in myoepithelial cells and intralumenal papillary protrusions of the tumor, whereas the antibodies to hepatic cytokeratin D presented an almost complementary pattern in that they showed strongest staining in the more basally located layers of tumor cells. Intraductal adenocarcinomas of rats showed strong staining with all keratin antibodies examined. In contrast to previous studies using exclusively antisera raised against epidermal prekeratin, out results show that all types of neoplastic and non-neoplastic epithelial cells of mammary gland of both species contain-at least some-filaments of the cytokeratin type identifiable by immunologic reaction, if antibodies are used that recognize a broad range of epidermal and nonepidermal cytokeratins. Consequently, such broad range antibodies to keratin-like proteins provide adequate tools to identify and characterize neoplastic and non-neoplastic epithelial cells and to eliminate false negative immunocytochemical findings in tumor diagnosis. In addition, our observation that in the same human carcinoma two cell types can be distinguished by their reaction with two different antibodies to cytokeratins from epidermis and liver, respectively, indicates that the cells of a given carcinoma can differ in their cytoskeletal composition, thus presenting further criteria for diagnostic differentiation.

Predictive animal testing for photocontact allergenicity.

The purpose of this investigation was to establish a standardized animal model to predict the photoallergenic potential of new chemical compounds. In a first series of experiments the main factors influencing the induction of photoallergenicity were evaluated (induction concentration, pH of the test solution, pretreatment of the irradiation site with sodium lauryl sulphate, additional use of adjuvant injections and routes of administration). Osram Ultravitalux lamps were utilized for these studies. In a second series of experiments a Xenon 6000 W radiation source was used. Positive results were obtained with well known photosensitizers, as, for instance, the halogenated salicylanilides, as well as with so-called weak photoallergens (e.g. bithionol, sulphanilamide) by stimulating the animals' immune system with Freund's complete adjuvant. The photoallergenic compounds were at the same time tested for their contact allergenic potential in the optimization test. A standard procedure for the predictive photoallergenicity assay in guinea-pigs is proposed.

Evaluation of the photocontact allergenic potential of 6-methylcoumarin in the guinea pig.

6-Methylcoumarin was tested in a new animal model developed for the predictive evaluation of photocontact allergenic potential. Negative results were obtained and these are discussed with respect to inherent limitations of the animal model.

The optimization test in the guinea pig in relation to other predictive sensitization methods.

The allergenicity of various compounds was tested by means of the optimization procedure in the guinea-pig. Results with antibiotics, preservatives, fragrance raw materials and miscellaneous contactants are reported. Their relevance is critically discussed in relation to other animal sensitization methods (Draize, Bühler, open epicutaneous and maximization tests). Predictive animal tests are compared with those used in human allergy (Shelanski & Shelanski; maximization test). The importance of animal methods entailing the use of adjuvants is underlined.

[Photocontact allergy. On the epidemiology and possible prediction from animal experiments (author's transl)]. / Photokontaktallergie. Zur Epidemiologie und prädiktiven tierexperimentellen Erfassung.

The incidence of photocontact allergic reactions in man is compared with that of pure contact allergic reactions by reviewing the literature of the last 15 years. A list of the more important photoallergenic compounds with the numbers of published clinical cases has been compiled. The common animal test procedures are reviewed and critically discussed.

[Effect of dermatocorticoids on the activity of hepatic tryptophane pyrrolase in the guinea pig (author's transl)]. / Wirkung von Dermatocorticoiden auf die Aktivität der hepatischen Tryptophanpyrrolase beim Meerschweinchen.

The enzyme activity of tryptophane pyrrolase in guinea pig liver can be measurably increased--as in other mammalian species--by systemic administration of corticoids, but this effect is short-lived and achieved only with high s.c. doses of, e.g., prednisolone (Ultracorten-H-hydrosoluble). In contradistinction and rather surprisingly the enzyme activity is variably but markedly reduced by high-dosed and protracted epicutaneous application (10 times within 2 weeks) of the well-known dermatocorticoids: hydrocortisone, fluocinolone acetonide, and clobetasol propionate. This reaction must be explained by the capacity of these corticoids to inhibit, after their percutaneous absorption, the functional axis pituitary--adrenal cortex--liver, the stimulatory role of which is mandatory for the endogenous basic activity of hepatic tryptophane pyrrolase.

Erythrocyte adherence to the marginal zone of mouse spleen follicle mediated by receptor(s) for neuraminic acid.

Erythrocytes of different species (chicken, sheep, man, mouse, rat, guinea pig) except rabbit erythrocytes strongly adhere to the marginal zone of mouse spleen follicles in frozen sections. This adherence reaction (AR) is not restricted to red blood cells but is also observed with human lymphocytes. Pretreatment of the tissue sections with trypsin, mercaptoethanol, periodate, chloroform/methanol, acetone, and heating the sections abolishes AR whereas neuraminidase (VCN) treatment of the sections has an amplifying effect. AR is inhibited by preincubation of the neuraminidase- or untreated sections with neuraminic acid (NA). Treatment of the erythrocytes with VCN completely abolishes AR whereas treatment with other enzymes (hyaluronidase, collagenase) is ineffective in this respect. Determination of NA in the erythrocyte membrane before and after VCN treatment reveals a positive correlation between the amount of NA and AR. Rabbit red blood cells have the lowest NA content in their membranes and, in addition, there is little effect of VCN treatment in further reducing it. It is possible that a lectin-like substance is responsible for AR. The biologic significance of AR is hypothetical, but since AR occurs in an area of the spleen playing a role in antigen trapping it is conceivable that this trapping may be mediated by interaction of NA and NA receptor(s).

Percutaneous absorption of clioquinol (Vioform).

The percutaneous absorption of clioquinol from three different preparations for skin treatment (Vioform cream, Locacorten-Vioform cream and Vioform-Hydrocortisone cream) was evaluated. After topical dosages corresponding to 30 mg clioquinol, concentrations in the blood were below the detection limit of the analytical procedure, i.e., smaller than 0.02 micrograms/ml; therefore the percutaneous absorption was evaluated by measuring cumulative urinary excretion of clioquinol and was compared to that found after an equivalent oral dose. The study was carried out in 4 healthy volunteers. The topical preparations were applied under occlusive dressings. Following epicutaneous application of the three topicals in quantities containing 30 mg clioquinol each, the urinary excretion of the drug was between 1.2 and 3.6% of the applied dose. When the same dose of clioquinol was administered orally to two volunteers, 52.4 and 92.9% of the dose was excreted in the urine. Taking the urinary elimination as the minimal amount of drug absorbed, the extent of percutaneous absorption from the three dermatological preparations amounted to 1.2-3.6% of the applied dose. There was no difference in the pattern of urinary excretion products among the three topicals and the oral formulation. The bulk of clioquinol was excreted as glucuronide (mean: 96 +/- 3%) and only a small fraction was excreted as sulfate (mean: 3.8 +/- 3%). A small amount of free clioquinol (1.1%) was measured in 1 subject only after the oral dose.

Percutaneous absorption of chlorquinaldol (Sterosan).

The percutaneous absorption of chlorquinaldol from three different preparations for skin treatment (Sterosan-hydrocortison ointment and cream, Sterosan ointment) was evaluated by measuring the cumulative urinary excretion of chlorquinaldol and was compared to that found after an equivalent oral dose. In the blood the concentrations were below the detection limit of the analytical procedure, i.e., smaller than 0.02 microgram/ml. The study was carried out in 3 healthy volunteers using a four-period change-over design. The topical preparations were applied under occlusive dressings. Following epicutaneous application of the three topicals in quantities containing 30 mg chlorquinaldol each, the mean urinary excretion of the drug amounted to 11.1 +/- 6.6, 19.5 +/- 3.5 and 13.0 +/- 2.5% of the applied dose. When the same dose of chlorquinaldol was administered orally, 67.6 +/- 9.5% of the dose was excreted in the urine. Taking the urinary elimination as the minimal amount of drug absorbed, the extent of percutaneous absorption from the three dermatological preparations varied between 4.2 and 23.5% of the applied dose. There was no difference in the pattern of urinary excretion products among the three topicals and the oral formulation. The bulk of chlorquinaldol (mean 98.0 +/- 0.1%) was excreted as sulfate (mean 1.3 +/- 0.3%) or unchanged drug (mean 0.8 +/- 0.1%).

Antimicrobial spectrum of triclosan, a broad-spectrum antimicrobial agent for topical application. II. Comparison with some other antimicrobial agents.

The antimicrobial activity of 5-chloro-2(2,4-dichlorophenoxy) phenol (triclosan, one of the active ingredients of Logamel, Ciba-Geigy) was compared in vitro with that of other antimicrobials exclusively or occasionally used as topical agents in dermatology: hexachlorophene, clioquinol, chlorquinaldol. gentamicin, neomycin, nystatin, econazole, clotrimazole and salicylic acid. Upon determination of the MICs for 53 strains of aerobic and anaerobic bacteria, yeasts and fungi, triclosan was found to display a high degree of activity against most of the test organisms and to have the broadest spectrum of chemotherapeutically significant antimicrobial activity of the substances tested.

Predictive evaluation in animals of the contact allergenic potential of medically important substances. II. Comparison of different methods of cutaneous sensitization with "weak" allergens.

Results of the optimization method and of other methods used to assess contact allergy in laboratory animals were compared with known epidemiological data on the occurrence of hypersensitivity reactions in man. Tests were performed with preservatives (formalin, ethylenediamine and sorbic acid), drugs (penicillin G, Benzocaine and sulphathiazole) and other contactants belonging to widely different chemical classes (p-phenylenediamine, triclosan, pyrazole derivatives, nickel and chrome salts, eugenol, isoeugenol and mercaptobenzothiazole). The degree of sensitization achieved in guinea pigs by the optimized procedure (intradermal test with adjuvant combination) and the maximization procedure was invariably superior to that produced by the epidermal method using prior irritation of the site of application. Both the optimized procedure and the maximization test seem to be capable of identifying contact allergens that cause hypersensitivity reactions in as few as 1 in 10,000 of the human population as a whole. The optimization test merits consideration as a standardized and efficiently predictive procedure.

Predictive evaluation in animals of the contact allergenic potential of medically important substances. I. Comparison of different methods of inducing and measuring cutaneous sensitization.

Groups of guinea pigs were sensitized with a 0.1% solution of dinitrochlorobenzene (DNCB) by the Draize intracutaneous method. The course of the induction process, the influence of the vehicles used and the extent to which the reactions are amenable to assessment according to objective criteria were examined. The sensitivity of the standardized Draize test was then compared with that of various other sensitization techniques, including: The intracutaneous test with adjuvant (optimization test) The maximization test according to Magnusson & Kligman (1969) The epidermal sensitization test The epidermal sensitization test with prior irritation of the contact site (by croton oil or sodium lauryl sulphate). Comparison of these methods revealed that either the additional application of adjuvant or prior irritation of the contact site augmented the degree of sensitization of DNCB just as greatly as the simultaneous use of adjuvant and prior irritation of the skin, (maximization test). The improved sensitization methods, and in particular the standardized optimization test, may prove to be of particular value for the study of so-called weak allergens.