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Influence of parallel vs. perpendicular source geometry on cathodic: arc efficiency and film quality for generation of ZnO thin films.

ZnO films have been grown on Si (111) substrates using a modified PAMBD (pulsed arc molecular beam deposition) reactive cathodic arc source employing either O2, N2, or NH3 as carrier gas. Utilizing new source geometry, a two to three fold improvement in source efficiency has been realized. Scanning electron microscopy analysis confirms that this new source configuration gives a significant reduction in marcoparticle contamination and exhibits good crystalline properties for room temperature deposition. ZnO films were grown with this new source and characterized using X-ray diffraction and X-ray photoelectron spectroscopy.

Development of a ribosomal DNA ITS2 marker for the identification of the thrips, Scirtothrips dorsalis.

The thrips Scirtothrips dorsalis Hood (Thysanoptera: Thripidae) is an invasive pest that poses a significant economical threat to U.S. agriculture and trade. In this study, DNA sequence data and polymerase chain reaction (PCR) were utilized to develop a molecular diagnostic marker for S. dorsalis. The DNA sequence variation from the internal transcribed spacer 2 (ITS2) region of nuclear ribosomal DNA (rDNA) was analyzed from various thrips species, including S. dorsalis. A primer set and polymerase chain reaction cycling parameters were designed for the amplification of a single marker fragment of S. dorsalis ITS2 rDNA. Specificity tests performed on ten thrips species, efficacy tests performed on fifteen S. dorsalis populations, and tests on primer sensitivity and robustness all demonstrated the diagnostic utility of this marker. This diagnostic PCR assay provides a quick, simple, and reliable molecular technique to be used in the identification of S. dorsalis.

Pyrazinoindolone inhibitors of MAPKAP-K2.

Optimization of pyrazinoindolone inhibitors of MAPKAP-K2 (MK2) provides a reasonable balance of cellular potency and physicochemical properties. Mechanistic studies support the inhibition of MK2 which is responsible for the sub-micromolar cellular efficacy.

Robotic assessment of locomotor recovery in spinal contused rats.

The purpose of this study was to investigate the ability of a robotic device, "the rat stepper", to assess intrinsic locomotor recovery following spinal cord contusion injury in adult rats. The device consists of a motorized body weight support mechanism that precisely controls the load to the hindlimbs during stepping, and two small robotic arms that measure and manipulate hindlimb movement. Sixteen rats received a contusion injury to the mid thoracic spinal cord with different severity levels (mild, moderate, severe, and sham). The animals were then evaluated weekly using the rat stepper, beginning one week after injury and continuing for a period of twelve weeks, across a range of body weight support levels. The contused animals demonstrated recovery in a standard locomotor assessment score (the BBB score), with most of the recovery occurring by four weeks post injury. We analyzed fourteen robotic measures of stepping and found that the measures that were most sensitive to intrinsic recovery were step velocity and inter limb coordination. These measures were also significantly correlated with the BBB score. The number of steps taken during testing was not sensitive to intrinsic recovery, nor correlated to the BBB score. These results suggest that step quality, rather than quantity, best reflects recovery after contusion injury in adult, untrained rats. Thus, robotic motion capture of only a few steps can provide a sensitive, valid measure of locomotor recovery after contusion.

Dynamic expression of L-selectin in cell-to-cell interactions between neutrophils and endothelial cells in vitro.

Neutrophil-endothelial cell interactions are regulated by cell adhesion molecules and their cognate ligands. It has been proposed that L-selectin and Mac-1 (CD11b/CD18), two neutrophil adhesion receptors, have sequential roles in neutrophil extravasation during inflammation. In this model, L-selectin mediates rolling and initial adherence of neutrophils to endothelial cells, while Mac-1 strengthens this initial adherence and also facilitates migration of neutrophils through endothelial cells. L-selectin and Mac-1 expression are known to be inversely regulated. Here an in vitro culture system has been developed to investigate in situ expression of L-selectin during cell-to-cell interactions between neutrophils and endothelial cell monolayers by confocal immunofluorescence analysis. Neutrophils underwent profound cell shape change from round to polarized cell morphology with pseudopod formation after 5 to 15 min coculture with IL-1-stimulated human endothelial cells. L-selectin was redistributed to the pseudopod of the polarized neutrophils in correlation with such cellular changes. During initial cell attachment, neutrophils bound to IL-1-stimulated endothelial cells expressed a high level of L-selectin in a polarized pattern. L-selectin expression decreased over time during neutrophil-endothelial cell interactions.

Detection of Chlamydia trachomatis by immunofluorescence, papanicolaou and immunoperoxidase in women with leucorrhea.

Bacteriological, Papanicolaou, direct immunofluorescense (DIF) and immunoperoxidase studies were done in 245 samples from women with vaginal discharge and the frequency of Chlamydia trachomatis was investigated. The samples were obtained from two hospitals of the Public Health Services of Mexico City. Samples were taken from the endocervical and posterior fornix areas and streaked in different cultures media for the isolation and microbiological differentiation. Smears were done and stained by Gram, Papanicolaou, DIF and immunoperoxidase techniques and the presence of cytological alterations and Chlamydia trachomatis was investigated. The microorganism was detected in 8 patients by the three methods, only 4 of these were found by DIF, 7 by cytological and 6 by immunoperoxidase techniques. The infection caused by C. trachomatis cytologically was associated with mild or moderate dysplasia but not with some special microorganism.

The role of combined pelvic floor stimulation and biofeedback in female urinary incontinence: early experience.

A series of 109 female patients with urinary incontinence were treated by pelvic floor stimulation and biofeedback. Eighty-one patients received the full course of treatment and 28 patients dropped out. Among the 81 patients who completed the treatment, the improvement and cure rates (completely dry) shortly after treatment were 70.4% and 12.3%, respectively. The improvement and cure rates for more than one year after treatment were 47.6% and 14.3% respectively. Seven patients returned for further pelvic floor stimulation and biofeedback and only three patients underwent bladder neck suspension operations after completing treatment. Of the patients who dropped out, 53.6% had shown improvement with pelvic floor stimulation and biofeedback. Side effects were rare with three patients complaining of abdominal pain. These findings suggest that pelvic floor stimulation and biofeedback are useful, safe and conservative treatment modalities for female patients with urinary incontinence.

Detection of hepatitis A virus and other enteroviruses in water by ssRNA probes.

Sensitive and specific methods are needed to detect hepatitis A virus (HAV) and other human enteroviruses in environmental samples such as drinking water and foods. Clones of cDNA encoding the 5'-most 1 kb of the HAV and coxsackievirus B3 (CB3) genomes were subcloned into T7/SP6 RNA transcription vectors. In vitro transcribed RNA from the T7 promoter detected their respective HAV or CB3 genomic RNA. Conversely, SP6 transcripts detected viral negative-stranded RNA but not the genome. When both ssRNA probes were tested at high temperature (65 degrees C), they did not hybridize with intracellular RNAs from 6 primate cell cultures used for isolation of HAV and other enteroviruses. The HAV probe did not hybridize with 13 different enteroviruses but detected as little as 500-1000 infectious units of the 7 strains of HAV tested. Conversely, the CB3 probe showed strong homology with all 13 enteroviruses tested but not HAV. The probes were used to detect HAV and other enteroviruses in water samples after virus amplification in cell culture. HAV was detected in water samples obtained during a waterborne hepatitis outbreak using the ssRNA probe. These samples were negative for HAV by direct solid phase radioimmunoassay and were not positive by immunoassays of inoculated cell cultures until several weeks of propagation. The CB3 ssRNA probe detected enteroviruses in samples of surface water and drinking water that were negative for cytopathic effects in inoculated cell cultures.

A competitive ELISA for the anti-intercellular adhesion molecule-1 (anti-ICAM-1) binding activity of monoclonal antibody R6.5 in serum.

A competitive ELISA was developed to measure serum concentrations of immunoreactive R6.5, a mouse IgG2a monoclonal antibody which binds to and inhibits the interactions of human intercellular adhesion molecule-1 with glycoproteins of the CD18 complex and with rhinoviruses. The assay design permitted quantitative measurement of the functional activity of the antibody in serum. The working range of the assay was from 2.15 to 215 ng ml-1, and intra-assay and inter-assay relative standard deviations averaged 13 and 20%, respectively. Serum caused interferences with the assay when spiked in vitro, but authentic serum samples obtained from intravenously dosed animals did not exhibit these interferences. Characteristics of the assay, as well as results of a pharmacokinetic study in rabbits, are presented.

Characterization of a receptor for human monocyte-derived neutrophil chemotactic factor/interleukin-8.

Monocyte-derived neutrophil chemotactic factor/interleukin-8 (MDNCF/IL-8) is an 8,000-dalton protein produced by monocytes which exhibits activity as a chemoattractant for neutrophils with maximal activity achieved at a concentration of 50 ng/ml. This polypeptide has been iodinated by chloramine-T methodology (350 Ci/mM), and specific receptors for MDNCF/IL-8 have been detected on human neutrophils, U937 cells, THP-1 cells, and dimethyl sulfoxide-differentiated HL-60 cells. The binding of MDNCF/IL-8 to human neutrophils is not inhibited by interleukin-1 alpha, tumor necrosis factor-alpha, insulin, or epidermal growth factor. In addition, chemoattractants such as C5a, fMet-Leu-Phe, leukotriene B4, and platelet-activating factor fail to inhibit binding, suggesting that MDNCF/IL-8 utilizes a unique receptor. The receptor for MDNCF/IL-8 is apparently glycosylated since ligand binding is inhibited by the presence of wheat germ agglutinin, a lectin with a binding specificity for N-acetylglucosamine and neuraminic acid. Steady state binding experiments indicate Kd values of 4 and 0.5 nM and receptor numbers of 75,000 and 7,400 for human neutrophils and differentiated HL-60 cells, respectively. 125I-MDNCF/IL-8 bound to human neutrophils is rapidly internalized and subsequently released from cells as trichloroacetic acid-soluble radioactivity. Affinity labeling experiments suggest that the human neutrophil MDNCF/IL-8 receptor exhibits a mass of approximately 58,000 daltons.

Generation of leukotrienes by antigen and calcium ionophore A23187 stimuli from a mouse mastocytoma cell line, MMC-16.

The generation of leukotrienes and histamine release by the mouse mastocytoma cell line MMC-16 was investigated. These cells produced leukotriene C4 (LTC4) and released histamine upon calcium ionophore A23187 and antigen stimulation. The ionophore also stimulated the biosynthesis of leukotriene B4 (LTB4) by MMC-16. Generation of LTC4 was confirmed by its characteristic UV absorption spectrum, fast atom bombardment-MS, equivalent HPLC retention time with an authentic standard and radioimmunoassay. Leukotriene B4 was characterized by its distinctive UV spectrum and HPLC retention time compared with synthetic material. IgE-mediated LTC4 generation was also observed in a dose dependent fashion with MMC-16 cells passively sensitized with monoclonal IgE specific for ovalbumin. LTC4 biosynthesis was effectively inhibited by the lipoxygenase inhibitor NDGA.

Pharmacological modulation of plasminogen activator secretion by P388D1 cell line.

P388D1 is a murine macrophage cell line which spontaneously secretes plasminogen activator (PA; activated function) and lysozyme (LYS; constitutive function). Compounds which decrease PA secretion without affecting LYS secretion have potential as "down-regulators" of macrophage function and, hence, of the immune system. Glucocorticoids (e.g., dexamethasone, IC50 less than 0.01 microM) and auranofin (IC50 = 1 microM) are positive in this model. In contrast, cyclooxygenase inhibitors (indomethacin, ibuprofen and piroxicam, all at 1 microM) boost PA secretion; lipoxygenase inhibitors (REV-5901, NDGA and piriprost, all at 10 microM) have little or no effect. Dexamethasone, but not auranofin, induces a urokinase-inhibitory activity which elutes between 0.13 and 0.19 M NaCl upon anion exchange HPLC (TSK-DEAE-5-PW). Fibrin overlay following SDS-PAGE of the HPLC peak reveals a urokinase-inhibitory band at approximately 90 Kd.

Phencyclidine during pregnancy in the rat: effects on locomotor activity in the offspring.

Either 5 or 10 mg/kg of phencyclidine hydrochloride (PCP) was administered by gastric intubation to gravid rats during the last two weeks of gestation. Intubation controls received the vehicle and all offspring were fostered to untreated controls at birth. PCP produced a decrement in maternal weight gain and a small but nonsignificant reduction in birth weight that was no longer evident at weaning. There were no maternal deaths nor were resorptions or stillbirths increased by PCP exposure. Offspring were tested for differences in locomotor activity from birth to weaning at 30 days of age and during adulthood. No behavioral differences were found among the preweanling or adult offspring. Results are compared with other prenatal studies of PCP toxicity and teratogenicity.