Interleukin-5 expression in the lung epithelium of transgenic mice leads to pulmonary changes pathognomonic of asthma.

We have generated transgenic mice that constitutively express murine interleukin (IL)-5 in the lung epithelium. Airway expression of this cytokine resulted in a dramatic accumulation of peribronchial eosinophils and striking pathologic changes including the expansion of bronchus-associated lymphoid tissue (BALT), goblet cell hyperplasia, epithelial hypertrophy, and focal collagen deposition. These changes were also accompanied by eosinophil infiltration of the airway lumen. In addition, transgenic animals displayed airway hyperresponsiveness to methacholine in the absence of aerosolized antigen challenge. These findings demonstrate that lung-specific IL-5 expression can induce pathologic changes characteristic of asthma and may provide useful models to evaluate the efficacy of potential respiratory disease therapies or pharmaceuticals.

The late, but not early, asthmatic response is dependent on IL-5 and correlates with eosinophil infiltration.

Early-phase reactions (EPRs) and late-phase reactions (LPRs) are characteristic features of bronchial asthma, although the pathogenetic mechanisms responsible for each of the responses are not fully defined. A murine model of EPRs and LPRs was developed to investigate the role of IL-5 and eosinophils in development of both responses. After initial intraperitoneal sensitization and airway challenge to ovalbumin (OVA), mice were provoked by additional exposure to OVA. An EPR, characterized by a transient increase in airway responsiveness, was observed 5-30 minutes after antigen provocation. This response was followed by an LPR that reached its maximum at 6 hours after challenge and was characterized by increased airway responsiveness and significant lung eosinophilia. The EPR was blocked by cromoglycate and albuterol, whereas the LPR was abolished by cromoglycate and hydrocortisone. Before provocation with allergen, administration of anti-IL-5 antibody prevented the influx of eosinophils into the lung tissue and abolished the LPR but not EPR. These results suggest that IL-5 and eosinophils are essential for development of the LPR, but not EPR, in this model.

Cloning of the murine eosinophil peroxidase gene (mEPO): characterization of a conserved subgroup of mammalian hematopoietic peroxidases.

The mouse eosinophil peroxidase (mEPO) gene was cloned by screening a random-primed bone marrow cDNA library at reduced criteria using a hEPO cDNA. An mEPO cDNA was subsequently used to isolate the mEPO gene from a lambda-genomic library. The mEPO gene displays a high degree of conservation with its human homologue: the transcription units are approximately the same size, conserve the relative size and position of the 12 exons associated with each gene, and at a nucleotide level the mouse and human EPO genes are 86% identical in the protein coding regions and 66% identical in the 3'-untranslated trailer regions. This strong conservation extends to the encoded proteins which show approximately 90% amino acid identity. Expression of the mEPO gene is restricted to tissues containing eosinophil progenitor cells (e.g., bone marrow and spleen), a pattern similar to the expression of another murine eosinophil granule protein, major basic protein.

Identification of a new murine eosinophil major basic protein (mMBP) gene: cloning and characterization of mMBP-2.

We have identified a new eosinophil major basic protein gene family member in the mouse and have given it the designation murine major basic protein-2 (mMBP-2). The gene was initially characterized as a unique expressed sequence tag (EST) clone having significant identity to the previously recognized member of this gene family, mMBP-1. The EST was used to screen and isolate mMBP-2 from a bone marrow cDNA library. In addition, a genomic clone of mMBP-2 was isolated and this gene was shown to be physically linked to within 100 kb of mMBP-1 on the central region of mouse chromosome 2. Progressive similarity alignment of the deduced mMBP-2 open reading frame demonstrates the apparent conservation of the "pre-pro-mature" protein structure found in the other known mammalian MBPs. Mature mMBP-2 maintains the cationic nature associated with these proteins with a predicted pI of 9.95. However, unlike the human MBPs, which display a three orders of magnitude charge difference [hMBP-1 (pI 11.4) vs. hMBP-2 (pI 8.7)], mMBP-2 is only slightly less cationic than mMBP-1 (pI 10.5). Expression studies demonstrate that transcription of the mMBP-2 gene parallels mMBP-1 and is confined to hematopoietic compartments engaged in eosinophilopoiesis. Moreover, using mMBP-1 knockout mice and immunohistochemistry with an antisera that recognizes both mMBP-1 and -2, we demonstrate that mMBP-2 protein expression is restricted to eosinophil lineage-committed cells.

Synthesis and biological activity of some antitumor benzophenanthridinium salts.

A facil synthesis of benzophenanthridinium salts has been developed and used for preparing a number of these compounds. The antitumor activities in mouse leukemia L1210 (LE) and P388 (PS) were determined as well as some selected antimicrobial activities. Although antitumor activity was exhibited by several of the derivatives, none was as active as the naturally occurring alkaloid fagaronine. Fagaronine was made available as a synthetic by an improved procedure. Some structure-activity relationships among antitumor benzophenanthridinium salts are discussed.

Soluble P-selectin glycoprotein ligand 1 inhibits ocular inflammation in a murine model of allergy.

PURPOSE: To assess the anti-inflammatory modality of a soluble extracellular form of P-selectin glycoprotein ligand 1 (sPSGL-1) in a mouse model of ocular allergic response. METHODS: Potential anti-inflammatory effects of sPSGL-1 were investigated in SWR/J mice sensitized by topical application of short ragweed pollen to the nasal mucosa followed by a challenge of the ocular mucosa with the same allergen. Five experimental groups were included in these studies: A, mice neither sensitized nor challenged with pollen (control group 1); B, animals sensitized but not challenged (control group 2); C, animals not sensitized but challenged (control group 3); D, animals sensitized and challenged; and E, sensitized animals treated with sPSGL-1 before pollen challenge. All experimental groups were evaluated for gross morphologic ocular changes, and histologic assessments were made to determine the onset/progression of inflammatory reactions and to look for evidence of eosinophil infiltration. RESULTS: Mice sensitized and challenged with pollen developed clinical signs consistent with human allergic conjunctivitis. These signs correlate with histologic changes in the conjunctival epithelium and stroma (e.g., edema and extensive eosinophil infiltration). Moreover, the ocular changes also correlated with evidence of eosinophil degranulation. However, sensitized and challenged mice concurrently treated with sPSGL-1 displayed no inflammatory ocular changes associated with a ragweed-induced type-1 hypersensitivity reaction. The lack of ocular changes included the absence of histologic late-phase inflammatory changes of the conjunctiva and a 97% reduction in the induced eosinophil infiltrate. CONCLUSIONS: The antagonistic intervention of cell- cell interactions through the blockade of selectin-dependent leukocyte adhesion may offer novel therapeutic strategies to modulate inflammatory responses. The potent inhibitory effects on eosinophil recruitment and late-phase inflammation suggest a role for sPSGL-1 in the treatment of ocular allergic diseases.

Evaluation of supercritical fluid extraction in the pharmaceutical industry.

Various steroids precipitate from supercritical CO(2) as very small particles but still retain their crystallinity. This suggests that SCFE in this industry could be utilized as a size reduction method as an alternative to milling. The application of SCFE as a separation technique is demonstrated by the simultaneous extraction and reaction of mevinolin, a metabolite of the fungus Aspergillus terreus.

Role of endoscopic retrograde cholangiopancreatography in the management of suspected choledocholithiasis.

BACKGROUND: Indications for endoscopic retrograde cholangiopancreatography (ERCP) are not precisely defined. With the increasing availability of magnetic resonance cholangiopancreatography (MRCP) and several prospective studies proving the accuracy of this modality, it is important to select appropriate criteria for invasive testing when common bile duct (CBD) pathology is suspected. METHODS: We reviewed 200 consecutive ERCPs performed for jaundice, pancreatitis, noninvasive visualization of CBD stones, radiologic evidence of CBD dilatation, elevated liver function tests, or biliary colic. Diagnostic and therapeutic yields are determined for each indication and various combinations thereof. RESULTS: Of the 180 ERCPs performed for suspected choledocholithiasis, 97 (53.8%) were performed for multiple indications, whereas 83 (46.2%) were performed for only one indication. One hundred two patients (56.6%) had CBD pathology, whereas 78 explorations (43.4%) were negative. If multiple indications for ERCP were present, the diagnostic yield was 85.6%. When there was only one indication, the diagnostic yield decreased to 25.3%. CONCLUSIONS: ERCP is an invasive procedure with significant morbidity that should be used selectively. MRCP may be a more appropriate initial evaluation of suspected CBD pathology in many patients.

Decreasing the incidence of osteoporosis-related injuries through diet and exercise.

Osteoporosis is the most common systemic bone disorder in the United States. It affects 15 million people--primarily women--causing thousands of injuries and deaths per year at a cost estimated at +3.8 billion annually. Two important factors in preventing osteoporosis are regular exercise and adequate calcium intake throughout life. Studies have shown that the average daily consumption of calcium by premenopausal and postmenopausal American women is between one-third and one-half that needed to maintain a positive calcium balance and prevent the loss of bone mass. This proposal elaborates the following specific ways that our health care and educational institutions can change these prospects: a screening program for women of all ages, to identify those most at risk for developing osteoporosis; an increase in the recommended daily dietary allowance for calcium; a public information campaign about osteoporosis, using television and radio; an investigation of the feasibility of calcium additives in the American diet; and the establishment of an organization to develop educational programs and monitor research in osteoporosis prevention. Now is the time to make the appropriate efforts to better the prospects for millions to enjoy a long and healthier life.

Effectiveness of a topical antifungal agent (clotrimazole) in dogs.

An infection of Trichophyton mentagrophytes was induced in 13 dogs, and of Microsporum canis, in 9 dogs. After the lesions became established, they were assigned to a group for clotrimazole treatment, miconazole treatment, or no treatment. Treatments were applied one daily for 28 days, using a double blind method. The lesions were evaluated by daily scoring of severity and by semiweekly culturing throughout the treatment period. Compared with untreated controls, the response to clotrimazole was significantly better from day 11 until the end of the study. The response to clotrimazole equaled or occasionally exceeded that to miconazole.

The sensory history of developmentally delayed children with and without tactile defensiveness.

This study investigated whether sensory history questionnaires differentiated between developmentally delayed children with and without tactile defensiveness and, in addition, which individual items were most discriminative. Interviews were conducted with 20 mothers of developmentally delayed children regarding their children's past and present behavioral responses to tactile and other sensory stimuli. The group of mothers with tactually defensive children reported a significantly greater number of positive responses to items previously determined to be indicative of tactile defensiveness than did the mothers of children without tactile defensiveness. An individual item analysis identified 11 items as discriminating most clearly between a group of children with tactile defensiveness and a group without tactile defensiveness. Some preliminary behavioral trends and implications for the use of sensory history questionnaires in parent counseling and in planning appropriate treatment programs are presented.

Detection of tumor-specific antigens in an equine sarcoid cell line.

Indirect immunofluorescence and lymphocyte cytotoxicity experiments demonstrated the presence of a tumor-specific antigen(s) on the surface of cells from an equine sarcoid cell line (Mc1). Autologous serum (taken from the horse from which the Mc1 cells were derived) and sera from three other sarcoid-bearing horses revealed a similar membrane immunofluorescence when reacted with Mc1 cells, indicating the existence of cross-reacting antibodies. Results of serum colony inhibition experiments indicate that these antibodies are not cytotoxic. Incubation of Mc1 cells with autologous lymphocytes resulted in a toxicity to Mc1 cells, thereby demonstrating a cell-mediated immune reaction to autologous tumor cells.