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An effective cleaning and sanitation protocol is important to mitigate disease outbreaks in poultry rearing facilities. This is especially important in cases of salmonellosis where there is a risk of disease transmission to humans. Compressed air foam systems (CAFS) may serve as an alternative carrier for foaming agents to clean and sanitize agricultural surfaces following an outbreak. The objective of this study was to evaluate the effects of a CAFS applied commercial firefighting foam (FF) and chlorine-based foaming cleaner (FC) in floor pen and caged quail rearing facilities with a history of salmonellosis. A firefighting foam concentrate (Phos-Chek WD881®) and foaming cleaner (Chlor-A-Foam XL®) were diluted in water and applied to floor pen and caged rearing facilities using a compressed air foam system. Total aerobes and cellular adenosine triphosphate (ATP) were quantified pre- and post-treatment using swabs. Both treatments significantly reduced aerobic bacteria in the floor pen and caged quail facilities (P< 0.05). The greatest reduction of 1.74 log10CFU/mL was reported in the floor pen facility following FC treatment. Microbial ATP levels were also significantly reduced by both the CAFS applied FF and the FC in the floor pen and caged quail facilities (P< 0.05). Treatment of floor pen facilities with the FF resulted in the greatest ATP reduction of 4,201 RLU. These data summarize the efficacy of CAFS applied foaming agents suggesting that a compressed air foam system may serve as a practical method to clean quail rearing facilities.

Abstract Aims--To investigate how parents report children's respiratory sounds on video compared to a clinical 'gold standard'. Methods--Five clinicians agreed on 10 video clips of children with audible breathing. These [...]

A rapid and effective means to clean and disinfect premises is needed by the poultry industry. We hypothesized that commercially available foaming disinfectants and cleaners applied via a compressed air foam system (CAFS) may be used to significantly reduce aerobic bacteria in a commercial caged layer complex. Six field trials were conducted to evaluate current industry cleaning and disinfection protocols and the proposed CAFS application. A commercially available chlorinated alkaline foaming cleaner (CHL/ALK) was applied in trials 1 and 2 by CAFS to one half of the house, and the other half of the house was not treated. The entire house was then washed with a high-pressure water rinse (HPWR). A commercially available peroxyacetic acid (PAA) in trials 3 and 4 or a 14% glutaraldehyde (HI GLUT)/2.5% quaternary ammonia (QAC) blended disinfectant in trials 5 and 6 was applied by CAFS to one half of a washed house. The remainder of the house was then treated with a 7% glutaraldehyde (LO GLUT)/26% QAC, which was spray-applied to cages by the integrator. Environmental swabs of drinker cups and cage floors were collected pre- and post-treatment to determine if aerobic bacteria were reduced. The HPWR and the CHL/ALK treatments did not consistently reduce bacteria on treated surfaces. Significant differences were observed with each of the CAFS applications of the PAA product, resulting in bacterial reductions of 1.83 to 2.27 log10cfu/sample. Although inconsistent, differences (P< 0.05) were observed with the spray application of the 0.4% (v/v) LO GLUT/QAC with bacterial reductions of 0.42 to 2.15 log10cfu/sample on both surfaces. The CAFS application of the 1.6% (v/v) HI GLUT/QAC resulted in bacterial reductions (P< 0.05) of 3.11 to 3.78 log10cfu/sample. These data suggest that the application of 3.0% (v/v) PAA or a 1.6% (v/v) HI GLUT/QAC via CAFS may be used to consistently and significantly reduce aerobic bacteria in a commercial layer complex.

Objective.To describe the landscape of well-being content inclusion across schools and colleges of pharmacy in the United States and Canada through identification of content implementation, incorporation, and assessment.

Background: Recombinant activated factor VIIa (rFVIIa) is used to treat bleeds in hemophilia patients with inhibitors. A subcutaneous formulation could potentially improve its half‐life and make it suitable for prophylactic treatment. Objectives: A study was conducted to determine the safety of subcutaneously administered rFVIIa in patients with hemophilia and the pharmacokinetic profile (including bioavailability). Patients/Methods: This was a multicenter, open‐label, cross‐over comparison of single doses of intravenous rFVIIa 90 μg kg−1and a new formulation of rFVIIa for subcutaneous injection at dose levels of 45, 90, 180, 270 and 360 μg kg−1. Sixty subjects (12 per dose cohort) with hemophilia A or B were enrolled. Results: Subcutaneously administered rFVIIa showed lower mean peak plasma concentrations and prolonged FVII activity (Cmax, 0.44–5.16 IU mL−1[across doses]; t1/2, 12.4 h; tmax, 5.6 h) compared with intravenously administered rFVIIa (Cmax, 51.7 IU mL−1; t1/2, 2.7 h; tmax, < 10 min). The absolute bioavailability of subcutaneous rFVIIa ranged from 21.1 to 30.1% across dose levels. Dose proportionality was observed within a 2‐fold dose increase but not across the full dose range. No thromboembolic events, drug‐related serious adverse events, severe injection‐site reactions or neutralizing antibodies were reported (primary endpoint). Mild and moderate injection‐site reactions were more frequent with subcutaneous than with intravenous injections. Conclusion: This phase I clinical trial did not identify safety concerns of prolonged exposure to rFVIIa administered subcutaneously in single doses to hemophilia patients.

Abstract:: Haematological and serum biochemical data have been collected from a total of 104 tammar wallabies maintained in a captive population at Macquarie University, NSW, Australia. These data have been analysed with respect to the effects of age, sex and seasonality. Age-related effects were apparent for both haematological parameters and serum biochemistry. Erythrocyte parameters, including haemoglobin, total erythrocyte count, haematocrit, macrocyte volume index and mean corpuscular haemaglobin, but not the leucocyte parameters, total leucocyte count, neutrophils, lymphocytes, monocytes, eosinophils, platelets and maximum packed volume, differed between the sexes. Seasonal effects were apparent in both leucocyte and erythrocyte parameters. Serum parameters of total protein and bilirubin were higher in autumn whereas alkaline phosphatase values were higher in summer. These data will serve as an effective management tool for assessing and monitoring the health status of individuals, particularly those in captivity.

The female sex hormone 17β-estradiol (E2) has been shown to increase lipid and decrease carbohydrate utilization in animals. We administrated oral E2and placebo (randomized, double blind, crossover) to eight human male subjects for 8 days (∼3 mg/day) and measured respiratory variables, plasma substrates, hormones (E2, testosterone, leptin, cortisol, insulin, and catecholamines), and substrate utilization during 90 min of endurance exercise. [6,6-2H]glucose and [1,1,2,3,3-2H]glycerol tracers were used to calculate substrate flux. E2administration increased serum E2(0.22 to 2.44 nmol/l, P< 0.05) and decreased serum testosterone (19.4 to 11.5 nmol/l, P< 0.05) concentrations, yet there were no treatment effects on any of the other hormones. Glucose rates of appearance (Ra) and disappearance (Rd) were lower, and glycerol Ra-to-Rdratio was not affected by E2administration. O2uptake, CO2production, and respiratory exchange ratio were not affected by E2; however, there was a decrease in heart rate (P< 0.05). Plasma lactate and glycerol were unaffected by E2; however, glucose was significantly higher (P< 0.05) during exercise after E2administration. We concluded that short-term oral E2administration decreased glucose Raand Rd, maintained plasma glucose homeostasis, but had no effect on substrate oxidation during exercise in men.

A protocol is described to attach expression patterns to genes represented in a collection of hybridization array experiments. Discrete values are used to provide an easily interpretable description of differential expression. Binning cutoffs for each sample type are chosen automatically, depending on the desired false-positive rate for the predictions of differential expression. Confidence levels are derived for the statement that changes in observed levels represent true changes in expression. We have a novel method for calculating this confidence, which gives better results than the standard methods. Our method reflects the broader change of focus in the field from studying a few genes with many replicates to studying many (possibly thousands) of genes simultaneously, but with relatively few replicates. Our approach differs from standard methods in that it exploits the fact that there are many genes on the arrays. These are used to estimate for each sample type an appropriate distribution that is employed to control the false-positive rate of the predictions made. Satisfactory results can be obtained using this method with as few as two replicates.

This overview has been supported by a review of the literature referring to the management of community-acquired pneumonia (in preparation). Difficulties diagnosing bacterial pneumonia include identifying the pathogens and the validation of radiographic signs suggesting bacterial or mycoplasmal infection. The World Health Organisation (WHO) has published guidelines for diagnosis which seem to be as relevant for developed as developing countries. The main diagnostic features are tachypnoea, fever greater than 38.5°C and chest recession without wheeze.

BACKGROUND: Reactive oxygen and nitrogen derived species produced by activated neutrophils have been implicated in the damage of mucosal proteins including the inhibition of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in the active inflammatory lesion in patients with inflammatory bowel disease (IBD). This study investigated the efficacy of currently used IBD therapeutics to prevent injury mediated by reactive oxygen and nitrogen derived species. METHODS: GAPDH activity of human colon epithelial cells was used as a sensitive indicator of injury produced by reactive oxygen and nitrogen derived species. HCT116 cells (10(6)/ml phosphate buffered saline; 37 degrees C) were incubated in the presence of 5-aminosalicylic acid (5-ASA), 6-mercaptopurine, methylprednisolone, or metronidazole before exposure to H2O2, HOCl, or NO in vitro. HCT116 cell GAPDH enzyme activity was determined by standard procedures. Cell free reactions between 5-ASA and HOCl were analysed by spectrophotometry and fluorimetry to characterise the mechanism of oxidant scavenging. RESULTS: GAPDH activity of HCT116 cells was inhibited by the oxidants tested: the concentration that produced 50% inhibition (IC50) was 44.5 (2.1) microM for HOCl, 379.8 (21.3) microM for H2O2, and 685.8 (103.8) microM for NO (means (SEM)). 5-ASA was the only therapeutic compound tested to show efficacy (p&lt;0. 05) against HOCl mediated inhibition of enzyme activity; however, it was ineffective against H2O2 and NO mediated inhibition of GAPDH. Methylprednisolone, metronidazole, and the thiol-containing 6-mercaptopurine were ineffective against all oxidants. Studies at ratios of HOCl:5-ASA achievable in the mucosa showed direct scavenging to be the mechanism of protection of GAPDH activity. Mixing 5-ASA and HOCl before addition to the cells resulted in significantly greater protection of GAPDH activity than when HOCl was added to cells preincubated with 5-ASA. The addition of 5-ASA after HOCl exposure did not restore GAPDH activity. CONCLUSIONS: Therapies based on 5-ASA may play a direct role in scavenging the potent neutrophil oxidant HOCl, thereby protecting mucosal GAPDH from oxidative inhibition. These findings suggest that strategies for the further development of new HOCl scavenging compounds may be useful in the treatment of IBD.

Measurement of cellular DNA content by flow cytometry demonstrated presence of two distinct aneuploid neoplasms in a patient who developed acute myeloblastic leukemia (AML) 4 mo after diagnosis of a diffuse histiocytic lymphoma (DHL). A lymph node aspirate contained peroxidase- negative, “null,” hyperdiploid (2.6C) DHL cells, while the bone marrow (BM) contained 84% primitive peroxidase-positive tetraploid AML cells (4.0C). Minor populations of hyperdiploid HDL and normal diploid cells could be detected by flow-cytometry in the BM, and all three populations were also seen in the peripheral blood.

Bone marrow smears of 263 protocol patients with acute nonlymphoblastic leukemia (ANLL) and related disorders treated between 1970 and 1978 at MSKCC were reviewed blindly by two pairs of hematomorphologists and classified according to the FAB system. It was found necessary to add one category (MO) for acute undifferentiated leukemia and to define more precise quantitative criteria for the categories M1-M6 based on bone marrow differential counts. Using this modified FAB classification, agreement between the two observer groups based on morphology alone was 69%. Cytochemical stains were essential in establishing the diagnosis in 9%, led to a change of diagnosis by one observer team in 14%, and helped to confirm the diagnosis in 32% of cases. Complete remission rates, remission duration, and survival were not significantly different among diagnostic categories. Myelodysplastic syndromes (MDS: M6, RAEB, CMML; CR rate 48%) and ANLL without differentiation (M0, M1, M5a; CR rate 50%) appeared to do less well than ANLL with partial differentiation (M2, M3, M4, M5b; CR rate 59%) on all three protocols studied. Auer rods were present in 53% of all cases with 63% in the myeloid categories (M1-M4). Auer rods were found to be the single most important prognostic parameter in this study, with a complete remission (CR) rate of 68% in the Auer-rod- positive and of 40% in the Auer-rod-negative group (p < 0.0001). Survival was significantly longer for patients exhibiting Auer rods (p < 0.0002). Median survival for the total group was 13.5 mo and median remission duration for responders was 11.5 mo in the Auer-rod-positive group compared to 6.2 mo median survival and 9.2 mo median remission duration for the Auer-rod-negative group.

Five cases of adult leukemia with L3 morphology in bone marrow were studied for the presence of immunologic, metabolic, and enzymatic markers. Among the five patients, there were four males and female. Median age was 66 with a range of 16–80 yr. Median survival was only 5 mo. Serum lactate dehydrogenase (LDH) levels, 3H-thymidine labeling indices, and DNA/RNA content of the L3 lymphoblasts were markedly elevated. B-cell markers were found in three cases, two exhibiting surface membrane IgM-lambda, and one IgG-K. Terminal deoxynucleotidyl transferase (TdT) enzymatic activity was consistently low in this group. In one case, the L3 lymphoblasts displayed only surface Fc receptors demonstrated by the binding of aggregated IgG. TdT activity was found to be significantly increased. In another instance, the lymphoblasts formed spontaneous rosettes with sheep erythrocytes and exhibited paranuclear staining with acid phosphatase. TdT activity was found to be low. Although most of the L3 leukemias are neoplasias of B lymphocytes, other lineages may also express this morphology.

Leukemic cells from nine cases of acute monocytic leukemia (AMoL) were characterized by multiple differentiation markers. Cells in most cases were phagocytic, carried an Fc receptor, and stained positively for alpha-naphthyl acetate esterase but negatively for naphthol AS-D chloroacetate esterase. However, subtle differences in marker expression were observed which suggested different degrees of leukemic cellular maturation or activation. Cell marker analysis proved to be a useful adjunct to conventional morphology in confirming the diagnosis and the recognition of the neoplastic cells in AMoL, and may ultimately provide insight into the functional state of these cells.

Heat-stable enterotoxins (ST-124 and ST-1261) have been isolated from two different enterotoxigenic Escherichia coli of bovine (124) and porcine (1261) origin. The enterotoxin preparations were isolated by ultrafiltration and ion-exchange chromatography and were both active in the suckling mouse test and pig ligated loop test in the nanogram range. The bovine (ST-124) enterotoxin was not stable to heating in its isolated form, and significant differences in amino acid composition were observed between the two enterotoxins. Although both toxins were active at similar levels in the suckling mouse and pig ligated loop tests, ST-124 lacked the ability to cause the profound secretory responses seen with ST-1261 in the weanling pig ligated loop.

Southern blotting using radiolabeled probes is a well established technique for the detection of the Philadelphia translocation in the diagnosis of chronic myelogenous leukemia (CML). However, the use of radioisotopes in the clinical setting is often problematic. Because of this we investigated the use of a digoxigenin-labeled probe and chemiluminography in the detection of the Philadelphia translocation. In this study DNA was extracted from 19 bone marrow or blood samples from patients with CML or other malignancies and subjected to Southern blotting with a probe specific for the Philadelphia translocation, Phl/bcr3. The probe was labeled with either, 32P or digoxigenin to determine the relative sensitivity and specificity of autoradiography and chemiluminography in the molecular diagnosis of the BCR/abl fusion gene. All 19 samples were tested by both methods. All blots were performed and interpreted by individuals blind to the initial patient diagnosis. In addition, 12 samples (6 positive for CML, 6 negative for CML as determined by Southern blotting with 32P-labeled probe) were subjected to triplicate Southern-blot analyses, with three separate lots of digoxigenin-labeled probe to assay batch to batch variability in the efficacy of the probe. Radiolabeled and digoxigenin-labeled probes resulted in identical diagnoses in all cases. All results obtained by molecular analysis correlated perfectly with the clinical diagnoses of the patients from whom the samples had been obtained. Preanalysis of patient samples with different batches of digoxigenin-labeled probe gave highly reproducible results. With digoxigenin-labeled probe, diagnostic results were obtained after exposure times of less than 1 h at room temperature. Comparable results with radiolabeled probe required the use of enhancing screens and a 3–4-day exposure at −70°C. Overall, we have found the use of the digoxigenin-labeled probe Phl/bcr3 and chemilumines-cence using the substrate CSPD to be at least as good and in some respects superior to the use of radiolabeled probe and autoradiography in the molecular diagnosis of CML.

This paper used the standardized criteria proposed by Aylward et al I995 for diagnosing Alzheimer's Disease in individuals with learning disabilities to undertake a prospective, longitudinal study of 22 individuals with Down's syndrome. Results show that, after a period of up to 6 years, four individuals showed functional decline. A similar pattern of decline was found to that described in previous studies, although the mean age of onset was later. All four subjects met the criteria for probable Alzheimer's Disease.

4'-Demethoxydaunorubicin (idarubicin [IDR]) is a new anthracycline that differs from its parent compound by the deletion of a methoxy group at position 4 of the chromophore ring. This minor structural modification results in a more lipophilic compound with a unique metabolite that has a prolonged plasma half-life as well as in vitro and in vivo antileukemia activity. To determine its activity in acute myelogenous leukemia (AML), 130 consecutive adult patients between the ages of 16 and 60 with newly diagnosed disease were randomized in a single institution study to receive either IDR in combination with cytosine arabinoside (Ara-C) or standard therapy with daunorubicin (DNR) and Ara- C. The trial was analyzed using the O'Brien-Fleming multiple testing design that allowed for periodic inspection of the data at specific patient accession points. After accrual of 60 patients per arm, analysis showed that patients who received IDR/Ara-C had a superior response compared with those who received standard therapy: 48 of 60 patients (80%) achieved complete remission on the former arm compared with 35 of 60 patients on the latter (58%, P = .005). Logistic regression analysis of factors associated with complete response indicated that treatment with IDR/Ara-C offered a significant advantage to patients who presented with a high initial white blood cell count compared with treatment with DNR/Ara-C. The degree of marrow aplasia was approximately the same on each arm as was nonhematologic toxicity. Overall survival for patients on the IDR/Ara-C arm was 19.5 months compared with 13.5 months on the DNR/Ara-C arm (P = .025) at a median follow-up of 2.5 years. We conclude that IDR/Ara-C can effectively replace standard therapy with DNR/Ara-C in adult patients less than age 60 with newly diagnosed AML.

Major histocompatibility (MHC)-restricted, human immunodeficiency virus type one (HIV-1)-specific, cytotoxic T lymphocytes (CTLs) were detected in the peripheral blood mononuclear cells (PBMCs) of HIV-1-infected individuals. Using a system of autologous B and T lymphoblastoid cell lines infected with recombinant vaccinia vectors (VVs) expressing HIV-1 gene products, we were able to detect HIV-1-specific cytolytic responses in the PBMCs of 88% of HIV-1-seropositive hemophiliac patients in the absence of in vitro stimulation. These cytolytic responses were directed against both HIV-1 envelope and gag gene products. The responses were resistant to natural killer (NK) cell depletion and were inhibited by monoclonal antibodies (MoAbs) to the T cell receptor, CD8 surface antigens, and MHC class I antigens, suggesting a classical MHC class I restricted, virus-specific CTL response.

Leukemic cells in 134 patients with ALL were analyzed by a panel of mouse monoclonal antibodies. Two antibodies are reactive with all peripheral blood T cells but define different surface antigens (Leu-1 and Leu-4). Two other antibodies react with antigens that are restricted to suppressor/cytotoxic T cells (Leu-2) and to helper T cells (Leu-3). We also used antibodies to the receptor for sheep red blood cells (SRBC) (Leu-5) and to a human “TL-like” antigen that is found on most thymocytes but not in peripheral T cells (Leu-6). An antibody to the human p29.34 “Ia-like” molecule was also tested. Of the 134 ALL patients, 17 had a predominance of SRBC-rosetting (Leu-5+) lymphoblasts (“T” ALL), expressing different surface phenotypes defined by this panel of monoclonal antibodies. These phenotypes were not readily classifiable according to a scheme of sequential stages of normal differentiation proposed. Moreover, the lymphoblasts in 8 of 113 patients not expressing conventional B- to T-cell markers (“null” ALL) reacted with the monoclonal anti-T-cell antibodies. This study suggests that the classification of lymphoblasts in ALL based on the reactivities observed with this panel of mouse monoclonal antibodies is not easily reconciled with current models of normal T-cell differentiation. However, it should be emphasized that the precise sequence of antigenic expression by cells undergoing thymic differentiation is still not fully known, and further phenotypic analysis of ALL cells might contribute to an improved understanding of this malignancy.

Two distinct cell populations with lymphoblastic and monocytic characteristics were separated and characterized by multiple cell markers in a patient with terminal transferase-positive acute acute leukemia. The clinical course and sequential cell marker studies were consistent with the interpretation of a defect at the level of a common stem cell giving rise to a terminal transferase--positive lymphoblastic cell population at diagnosis and, following initial therapy, a terminal transferase--negative monocytic population.

Seventy-four patients presenting with a mixed pattern of urinary symptoms were randomly allocated to undergo either inpatient or outpatient continence programs as initial treatment, without prior urodynamic investigation. Both programs consisted of physiotherapy, bladder retraining, fluid normalization, dietary advice and general support and advice. Nine out of 39 in the outpatient group and 8 out of the 35 of the impatient group failed to complete the study. There was a significant decrease in frequency, nocturia, number of incontinent episodes and visual analog scores for both groups. In addition the outpatients had a significant reduction in loss on pad testing, and a significantly greater improvement in their visual analog score. In each group 63% were cured or improved to the extent that they did not require further treatment. Staff costs per outpatient were half those for an inpatient. We conclude that outpatient conservative treatment as detailed above is a successful first-line treatment of urinary incontinence in women. It is as successful and possibly better than inpatient treatment, and is significantly cheaper.

Fc gamma receptors provide an essential link between cellular and humoral immunity, and little is known about their expression in monocytes of newborn infants. We compared baseline and gamma interferon (IFN-gamma)-induced expression of Fc gamma RI and Fc gamma RII protein and Fc gamma RI mRNA in monocytes from healthy, term infants and adults. Fluorescence-activated cell sorter analysis demonstrated that baseline expression of monocyte Fc gamma RI in newborn infants was not significantly different from that in adults, while Fc gamma RII protein expression in monocytes derived from newborns was significantly higher than that for adults (mean channel fluorescence [MCF] for newborns and adults, 5.53 and 4.50, respectively [P = 0.039]). In vitro treatment with recombinant IFN-gamma increased the expression of Fc gamma RI in monocytes of newborns and adults to the same extent (2.4- and 2.2-fold increase in MCF in newborns and adults, respectively, at 42 h). We developed a semiquantitative fluorescence reverse transcriptase PCR which demonstrated a significant increase in mRNA for Fc gamma RI in monocytes of newborns and adults with in vitro IFN-gamma exposure, indicating that IFN-gamma acts by increasing the transcription or transcript stability of Fc gamma RI mRNA. While there was no significant effect of IFN-gamma treatment on Fc gamma RII expression in monocytes from adults, there was a 20% increase in Fc gamma RII in monocytes from newborns (P = 0.009). Monocytes from healthy, term newborns and adults exhibit comparable baseline and IFN-gamma-induced levels of expression of Fc gamma RI and higher baseline and IFN-gamma-induced levels of expression of Fc gamma RII.

Heat treatment of D. melanogaster tissue culture cells causes drastic changes in the pattern of protein synthesis and the size distribution of polysomes. Like the heat shock puffs on polytene chromosomes which appear while preexisting puffs regress, heat shock proteins appear on gels while the synthesis of preexisting proteins is sharply reduced, and heat-induced polysomes appear on gradients after preexisting polysomes have disappeared. Most of the poly(adenylic acid)-containing RNA isolated from high-temperature polysomes sediments in sucrose gradients and migrates in gels as a rather narrow band. This RNA is of sufficient size to code for one particular protein that is found to account for more than half of the total synthesis at high temperature. The RNA hybridizes in situ mainly at chromosome sub-division 87B, the site of the major heat shock puff.

The management of a chronic injury to an international squash player is described. By good physiotherapeutic management and the involvement of an interdisciplinary team it was possible to make gains in the non-playing period. The rehabilitation period included physiological assessment on the basis of which a training programme was devised with target goals set throughout the period. Mental rehearsal of skills was included at all stages. As a result, the player's confidence was maintained and fitness levels improved. The long term prognosis has been good. The model could be applied to any sports injury.

During the 12 year period, July 1980 to June 1992, 17 patients with blunt splenic trauma, aged 3.5 to 13.2 years (median 7.5), were treated. Ultrasound scanning was found to be valuable for diagnosis as well as follow-up of the healing of splenic injury. Eleven patients were observed on bed-rest without surgical intervention. The remaining six patients underwent exploratory laparotomy for suspected continuing intra-abdominal haemorrhage or because associated injuries rendered assessment of the possibility of intra-abdominal bleeding unreliable. In each case the spleen was conserved. There was no mortality or late complication.

We have constructed a vaccinia virus recombinant that expresses the extracellular domain of the rat neu oncogene-encoded protein, a 185-kDa transmembrane glycoprotein termed p185. Strain NFS mice immunized with this recombinant virus developed a strong antibody response against the neu oncogene product and were fully protected against subsequent tumor challenge with neu-transformed NIH 3T3 cells. No tumor immunoprotection was found when recombinant virus-immunized mice were challenged with Ha-ras-transformed NIH 3T3 cells. These data indicate that immunization with a single oncogene-encoded antigen can fully and specifically protect animals against tumor cells bearing this antigen.

Antibody-dependent cell-mediated cytotoxicity (ADCC) specific for human immunodeficiency virus (HIV) has been described for HIV-infected individuals. To determine the antigenic specificity of this immune response and to define its relationship to the disease state, an ADCC assay was developed using Epstein-Barr virus-transformed lymphoblastoid cell line targets infected with vaccinia virus vectors expressing HIV proteins. The vaccinia virus vectors induced appropriate HIV proteins (envelope glycoproteins gp160, gp120, and gp41 or gag proteins p55, p40, p24, and p17) in infected lymphoblastoid cell lines as demonstrated by radioimmunoprecipitation and syncytia formation with c8166 cells. Killer cell-mediated, HIV-specific ADCC was found in sera from HIV-seropositive but not HIV-seronegative hemophiliacs. This HIV-specific response was directed against envelope glycoprotein but was completely absent against target cells expressing the HIV gag proteins. The ADCC directed against gp160 was present at serum dilutions up to 1/316,000. There was no correlation between serum ADCC titer and the stage of HIV-related illness as determined by T-helper-cell numbers. These experiments clearly implicated gp160 as the target antigen of HIV-specific ADCC activity following natural infection. Vaccines which stimulate antibodies directed against gp160, which are capable of mediating ADCC against infected cells, could be important for protection against infection by cell-associated virus.

Eight Mycobacterium tuberculosis and M. leprae genes were inserted into the vaccinia virus genome by in vivo recombination. The resulting virus recombinants were shown to express five different M. tuberculosis proteins (71, 65, 35, 19, and 12 kDa) and three M. leprae proteins (65 and 18 kDa and a biotin-binding protein) by Western immunoblot analysis, radioimmunoprecipitation, or black-plaque assay. When injected into BALB/c mice, the recombinants expressing the M. tuberculosis 71-, 65-, or 35-kDa protein and the M. leprae 65-kDa protein or the biotin-binding protein elicited antibodies against the appropriate M. tuberculosis or M. leprae protein. These vaccinia virus recombinants are being tested for the ability to elicit immune protection against M. tuberculosis or M. leprae challenge in animal model systems. The recombinants are also useful in generating target cells for assays aimed at elucidating the cellular immune responses to mycobacterial proteins in leprosy and tuberculosis. Furthermore, the M. tuberculosis 65-kDa protein and four of the other mycobacterial proteins share homology with known eucaryotic and procaryotic stress proteins, some of which may play a role in autoimmunity.

Total internal reflection (TIR)/fluorescence recovery after photobleaching (FRAP), which has been used to study adsorption and surface diffusion of proteins, was modified and applied to study DNA oligonucleotides at liquid/solid interfaces. Conventional TIR/spot FRAP and TIR/pattern FRAP techniques use a photomultiplier tube (PMT) to reveal the adsorption dynamics and surface diffusion rates of biomolecules, respectively. However, they do not provide spatial information on these interfacial processes. In this work, a cooled charge-coupled device camera is substituted for the PMT normally used. Studies of adsorption and surface diffusion of the well-characterized protein bovine serum albumin (BSA) validated the system's operation. Then, the desorption rate constant for a fluorescently tagged 21-mer DNA oligonucleotide (MW 7140 Da) was determined by spot FRAP. The desorption rate constants for strongly and weakly adsorbed oligonucleotides from (3-aminopropyl)triethoxy silane (APTES) glass were determined to be 0.02 and 0.19 s^-1, respectively. These are of the same order of magnitude as those for BSA (MW 67 000 Da) on APTES glass. The surface diffusion coefficients of oligonucleotide are approximately the same as those for BSA and are dependent on the surface concentration of the molecules on APTES-coated glass. Since the molecules differ by a factor of 10 in molecular weight, these results suggest that the shape of a adsorbate molecule and the strength of adsorbate/substrate interactions play a strong role in interfacial adsorption and diffusion. The substitution of a methyl group in APTES for a hydrogen atom increased the desorption rate constants and surface diffusion coefficients significantly.

Groups of nude mice, with both hind footpads infected with 10(8) Mycobacterium leprae organisms, were treated with 4-week courses of different drug combinations. The effect treatment on each group was evaluated by subinoculating footpad homogenates from the treated mice into groups of normal and nude mice for subsequent regrowth, assessed 1 year later. A combination of rifampin (RMP) with clarithromycin (CLARI), minocycline (MINO), and ofloxacin (OFLO) resulted in the complete killing of M. leprae after 3 weeks of treatment. A combination of sparfloxacin (SPAR) and RMP also resulted in a similar bactericidal effect after 3 weeks of treatment. Other drug combinations showed variable effects. Very little or no effect was observed with any regimen if the treatment was given for less than 2 weeks. World Health Organization (WHO) multidrug therapy (MDT) given for 8 weeks was as effective as the two combinations described above. The results suggest that multidrug combinations consisting of RMP-OFLO (or SPAR)-CLARI (and/or MINO) are as effective as the WHO MDT for the treatment of experimental leprosy. Moreover, they imply that these combinations, which were found to be active in a 4-week experimental treatment protocol, could be administered as treatment to patients for a period of time shorter than the present 2-year regimen without a loss of effectiveness.

Background: Increasing prevalence of childhood obesity has resulted in an accelerating rate of referrals of overweight patients to pediatric clinics for exclusion of endocrine or metabolic etiologies. The exclusion of Cushing’s disease (CD) requires complex and potentially invasive investigations. Objective: to evaluate the sensitivity of accurate measurements of height, weight and body mass index (BMI) in discriminating between simple obesity and CD. Methods and patients: Height, weight and BMI were measured at diagnosis in 25 patients with CD; 14 males, 11 females, mean age 12.9 yr (6.4–17.8) and 41 patients with simple obesity (SO), defined as BMI >2.0 SD; 20 males, 21 females, mean age 9.4 yr (3.5–15.6). Results: Mean (±SE) BMI SDS in the CD patients was 2.41±0.5 and in the SO patients 3.71±1.3. Height SDS in the CD patients was −1.88±0.24 and in the SO patients 1.18±0.19 (p<0.05). The mean (±SE) BMI SDS to height SDS ratio was significantly decreased in the CD compared with the SO patients; −1.81±0.54 vs+0.90±1.17 (p<0.0001). Conclusions: Simple, accurate measurement of height and BMI SDS values provides a quick, and sensitive diagnostic discriminator in pediatric patients with CD or SO, thus potentially avoiding complex investigations.

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