Studies on Alternaria allergens. I. Isolation of allergens from Alternaria tenuis and Alternaria solani.

Extracts of Alternaria tenuis and Alternaria solani were separated into dialyzable (molecular weight less than 10,000) and non-dialyzable forms. The latter was further fractionated by gel filtration through Sephadex G-100 followed by ion-exchange chromatography on DEAE-cellulose. The dialyzable material was fractionated by gel filtration through Sephadex G-50. The allergenic activities of the fractions obtained from the A. tenuis extract was measured in vitro by the radioallergosorbent test assay and the allergenic potency was measured by radioallergosorbent test inhibition assay. Allergenic activity was detected in most of the non-dialyzable fractions, the majority of the activity being in the last G-100 fraction (MW approximately 20,000) which was predominantly protein in nature. The same component may be responsible for the activity found in the dialyzate and its first G-50 fraction since the immunodiffusion studies indicated that the last G-100 fraction has antigenic components in common with those of the first G-50 fraction. In addition, cross-reactions between A. tenuis and A. solani extracts show that the two species share common antigenic determinants.     

恶性疟原虫113肽抗原基因重组的减毒鼠伤寒沙门菌在家兔中免疫应答的研究

我们已经在减毒鼠伤寒沙门菌ＳＬ３２６１以融合蛋白的形式表达了人工合成的恶性疟原虫杂合１１３肽基因ＡＢ（ＧＺ－ＡＢ）。活菌以２×１０９ＣＦＵ经口服免疫新西兰家兔，用ＥＬＩＳＡ测定抗体水平，结果于首次免疫或加强免疫后都可检测到一定水平的特异性抗体。所免疫的家免可以诱发针对恶性疟原虫抗原及ＧＺ－ＡＢ的迟发性超敏反应（ＤＴＨ）。我们的研究表明，含有多个恶性疟原虫抗原表位的人工合成基因可以在减毒鼠伤寒沙门菌中表达，活菌可诱发家兔产生特异的体液免疫及细胞免疫，为恶性疟口服活菌苗的制备打下基础。     

Biological characterization of a novel biodegradable antimicrobial polymer synthesized with fluoroquinolones.

Biomaterial-related infections continue to represent a significant challenge to the medical community. Several approaches have been utilized to incorporate antimicrobial agents at the surface of implant devices in attempts to delay or eliminate the formation of biofilms. To date, most of these strategies have focused on drug conjugation or diffusion-limited systems for the delivery of such pharmaceutical agents. More recently, work has been presented on the feasibility of incorporating drugs into the backbone of polymers as a main-chain monomer. When sequenced into the backbone of the polymer with other monomers that are hydrolytically sensitive to enzyme-catalyzed breakdown, it is thought that drugs may be able to be selectively released. Specifically, degradable polyurethanes have been synthesized with fluoroquinolone antibiotics and have shown an ability to kill bacteria when released following degradation of the polymer chains by the macrophage-derived enzyme cholesterol esterase. However, specificity of the cleavage sites in the polymer was difficult to control. Since cholesterol esterase has specificity for hydrophobic moieties, it is desirable to alter the formulation of the polyurethanes to incorporate long hydrophobic monomers immediately adjacent to the ciprofloxacin molecule. Hence, the current study focuses on evaluating the enzyme-catalyzed degradation of a degradable polyurethane synthesized with 1,12 diisocyanatododecane as a substitute for 1,6 diisocyanatohexane, which was used in previous work. Validation of specific ciprofloxacin release and the generation of antimicrobial are shown. A preliminary cell study to assess the cytotoxicity of this biodegradable antibiotic polymer shows that the material has no observable effects on cell proliferation or cell membrane structure.     

Strategies to improve the signal and noise performance of active matrix, flat-panel imagers for diagnostic x-ray applications

A theoretical investigation of factors limiting the detective quantum efficiency (DQE) of active matrix flat-panel imagers (AMFPIs), and of methods to overcome these limitations, is reported. At the higher exposure levels associated with radiography, the present generation of AMFPIs is capable of exhibiting DQE performance equivalent, or superior, to that of existing film-screen and computed radiography systems. However, at exposure levels commonly encountered in fluoroscopy, AMFPIs exhibit significantly reduced DQE and this problem is accentuated at higher spatial frequencies. The problem applies both to AMFPIs that rely on indirect detection as well as direct detection of the incident radiation. This reduced performance derives from the relatively large magnitude of the square of the total additive noise compared to the system gain for existing AMFPIs. In order to circumvent these restrictions, a variety of strategies to decrease additive noise and enhance system gain are proposed. Additive noise could be reduced through improved preamplifier, pixel and array design, including the incorporation of compensation lines to sample external line noise. System gain could be enhanced through the use of continuous photodiodes, pixel amplifiers, or higher gain x-ray converters such as lead iodide. The feasibility of these and other strategies is discussed and potential improvements to DQE performance are quantified through a theoretical investigation of a variety of hypothetical 200 microm pitch designs. At low exposures, such improvements could greatly increase the magnitude of the low spatial frequency component of the DQE, rendering it practically independent of exposure while simultaneously reducing the falloff in DQE at higher spatial frequencies. Furthermore, such noise reduction and gain enhancement could lead to the development of AMFPIs with high DQE performance which are capable of providing both high resolution radiographic images, at approximately 100 microm pixel resolution, as well as variable resolution fluoroscopic images at 30 fps.     

Studies of cervical ripening in pregnant rats: effects of various treatments

The exact mechanisms that regulate cervical softening or ripening during pregnancy are not completely understood. The aim of this study was to estimate the effects of various agents on cervical softening during pregnancy in rats. Cervical resistance was examined after treatment with nitric oxide (NO) donors and inhibitors and different hormonal agents. Cervical resistance was significantly reduced (P< 0.05) in rats treated with the NO donors: sodium nitroprusside, molsidomine and prostaglandin E(2). However, treatments with the NO synthase (NOS) inhibitors N(omega)-nitro-L-arginine methyl ester (L-NAME) and L-N(6)-1-iminoethyl-lysine (L-NIL), or the prostaglandin synthesis inhibitor, indomethacin, significantly increased resistance (P<0.05). The antiprogesterone, onapristone, reduced cervical resistance and its effects were only partially blocked by the progesterone agonist, promegestone. Relaxin reduced cervical resistance and NOS inhibitors partially blocked the effect of relaxin. These studies demonstrate that NO regulates cervical ripening. Relaxin also softens the cervix and may act by stimulating NO synthesis. Progesterone seems important in the control of cervical ripening, but its role appears complex. NO and prostaglandin pathways may independently control ripening by acting in parallel or synergistically.     

Association of gastric epithelial apoptosis with the ability of Helicobacter pylori to induce a neutrophil oxidative burst

Both polymorphonuclear cell infiltration and increased epithelial apoptosis are seen in gastric mucosa in the presence of Helicobacter pylori infection. This study examined the association between bacterial ability to stimulate an oxidative burst in neutrophils and epithelial apoptosis. Biopsy specimens were obtained from 15 patients to detect apoptotic cells by the TUNEL method. H. pylori strains isolated from corresponding stomach biopsy samples were tested for the ability to stimulate an oxidative burst in human neutrophils. Neutrophils were isolated from healthy subjects without H. pylori infection and the oxidative burst was measured by flow cytometry with dichlorofluorescein diacetate. Stimulation with H. pylori increased both the percentage of activated cells and fluorescence intensity. There was a significant positive correlation between the number of epithelial apoptotic cells and fluorescence intensity. Increased neutrophil oxidative burst stimulated by H. pylori may play a role in enhanced gastric mucosal DNA damage and consequent atrophic gastritis and gastric cancer.     

An unusual expression of a squamous cell marker, small proline-rich protein gene, in tracheobronchial epithelium: differential regulation and gene mapping.

An unusual expression of a putative squamous cell marker, small proline-rich protein (spr1), in mucociliary epithelial cells of conducting airways was demonstrated in a serum-free culture system. A cDNA clone was isolated from the cDNA library of monkey tracheobronchial epithelial (TBE) cells by differential hybridization. This cDNA clone, MT5, exhibited 98% homology to a DNA sequence obtained from human keratinocytes treated with either UV light or phorbol esters (T. Kartasova et al., 1988, Mol. Cell. Biol. 8:2195-2230). The predicted peptide of MT5 is unusual for its high content of proline (29%), glutamine (18%), and cysteine (9%) and its repeated PKVPEPC units. The level of spr1 mRNA in cultured cells was inhibited more than 90% by vitamin A. In contrast, phorbol 12-myristate 13-acetate (PMA) stimulated the level of spr1 mRNA by 3- to 8-fold. This differential regulation coincided with the effects of these chemicals on the cornification of cultured TBE cells. Using MT5 as a probe, we have localized the tracheal spr1 gene on the human chromosome 1 by a Southern blot analysis using a panel of human-rodent somatic cell hybrid DNAs. The gene was further sublocalized to bands q22-23 by in situ hybridization.     

Characteristics of Achromobacter xylosoxidans bacteremia in northern Taiwan.

There were 40 cases of Achromobacter xylosoxidans bacteremia during a 6-year period in a medical center in northern Taiwan. Males outnumbered females (72.5% vs 27.5%). Common underlying diseases and conditions in these 40 bacteremic patients were malignancies (57.5%), central venous catheter implants (55%), surgery (35%), neutropenia (30%) and use of steroids (25%). Recurrent bacteremia occurred in 4 patients (10%), all of whom had a central venous catheter implant which was considered the most probable source of infection. More than one-quarter (27.5%) of the cases were community acquired. The mortality rate due to A. xylosoxidans bacteremia was 47.5%. Risk factors significantly associated with mortality were nosocomial acquisition and polymicrobial bacteremia. Disk susceptibility testing showed that these isolates were resistant to aminoglycosides and most cephalosporins but were susceptible to piperacillin, imipenem, ceftazidime and trimethoprim-sulfamethoxazole.     

Distinct types of T-cell help for the induction of a humoral immune response to Streptococcus pneumoniae

Studies have indicated that purified soluble polysaccharide antigens can elicit T cell-independent Ig responses in vivo, although these responses can be modulated by T cells in a noncognate manner. Relatively little is known, however, concerning the parameters that regulate polysaccharide-specific, as well as protein-specific, Ig isotype responses to an intact extracellular bacterium. Using the murine in vivo humoral response to intact Streptococcus pneumoniae as a model it can be shown that CD4+ T-cell receptor alphabeta+ T cells deliver help for both polysaccharide- and protein-specific Ig responses. However, these responses differ fundamentally in their mechanism of action.