Measurement of allergen-specific secretory IgA in stool of neonates, infants and toddlers by protection against degradation of immunoglobulins and allergens

Background and aims : Measurement of secretory immunoglobulin A (SIgA) level is important to monitor various disease conditions. However SIgA in gut mucosa is degraded by pancreatic proteases and proteolytic enzymes from enteric microbiota. Currently, there is no reliable quantitation method that measures allergen-specific SIgA levels in stool of neonates, infants and toddlers. Methods : Allergen-specific SIgA levels in stool of 10 healthy neonates, infants and toddlers aged 0 to 36 months were measured by our new allergen microarray with densely carboxylated arms on a glass slide chip. Results : Protease activities in stool of 3-day-old neonates were low and no degradation of SIgA, IgA and allergens was detected. However, immunofluorescence signals of SIgA, IgA and allergen on the chip were markedly reduced by stool extracts obtained from infants and toddlers aged more than one month in dose- and time-dependent manners. Such reduction was almost completely inhibited by serine protease inhibitors, phenylmethylsulfonyl fluoride (PMSF) and partly by leupeptin, but not by a variety of other protease inhibitors tested. Conclusion : Allergen-specific SIgA levels in stool of neonates, infants and toddlers under 36 months of age could be analyzed using protease inhibitors, including PMSF and leupeptin

A Uniform Analysis of the Ly-alpha Forest at z=0 - 5: V. The extragalactic ionizing background at low redshift

In Paper III of our series "A Uniform Analysis of the Ly-alpha forest at z=0 - 5", we presented a set of 270 quasar spectra from the archives of the Faint Object Spectrograph on the Hubble Space Telescope. A total of 151 of these spectra, yielding 906 lines, are suitable for using the proximity effect signature to measure J(\nu_0), the mean intensity of the hydrogen-ionizing background radiation field, at low redshift. Using a maximum likelihood technique and the best estimates possible for each QSO's Lyman limit flux and systemic redshift, we find J(\nu_0)= 7.6^+9.4_-3.0 x 10^-23 ergs s^-1 cm^-2 Hz^-1 sr^-1 at at 0.03 < z < 1.67. This is in good agreement with the mean intensity expected from models of the background which incorporate only the known quasar population. When the sample is divided into two subsamples, consisting of lines with z 1, the values of J(\nu_0) found are 6.5^+38._-1.6 x 10^-23 ergs s^-1 cm^-2 Hz^-1 sr^-1, and 1.0^+3.8_-0.2 x 10^-22 ergs s^-1 cm^-2 Hz^-1 sr^-1, respectively, indicating that the mean intensity of the background is evolving over the redshift range of this data set. Relaxing the assumption that the spectral shapes of the sample spectra and the background are identical, the best fit HI photoionization rates are found to be 6.7 x 10^-13 s^-1 for all redshifts, and 1.9 x 10^-13 s^-1 and 1.3 x 10^-12 s^-1 for z 1, respectively. This work confirms that the evolution of the number density of Ly-alpha lines is driven by a decrease in the ionizing background from z ~ 2 to z ~ 0 as well as by the formation of structure in the intergalactic medium. (Abridged)Comment: 71 LaTeX pages, 20 encapsulated Postscript figures, Accepted for publication in ApJ, Figure 4 available at http://lithops.as.arizona.edu/~jill/QuasarSpectra/ or http://hea-www.harvard.edu/QEDT/QuasarSpectra

Global Gene Expression Profiling in PPAR-γ Agonist-Treated Kidneys in an Orthologous Rat Model of Human Autosomal Recessive Polycystic Kidney Disease

Kidneys are enlarged by aberrant proliferation of tubule epithelial cells leading to the formation of numerous cysts, nephron loss, and interstitial fibrosis in polycystic kidney disease (PKD). Pioglitazone (PIO), a PPAR-γ agonist, decreased cell proliferation, interstitial fibrosis, and inflammation, and ameliorated PKD progression in PCK rats (Am. J. Physiol.-Renal, 2011). To explore genetic mechanisms involved, changes in global gene expression were analyzed. By Gene Set Enrichment Analysis of 30655 genes, 13 of the top 20 downregulated gene ontology biological process gene sets and six of the top 20 curated gene set canonical pathways identified to be downregulated by PIOtreatment were related to cell cycle and proliferation, including EGF, PDGF and JNK pathways. Their relevant pathways were identified using the Kyoto Encyclopedia of Gene and Genomes database. Stearoyl-coenzyme A desaturase 1 is a key enzyme in fatty acid metabolism found in the top 5 genes downregulated by PIO treatment. Immunohistochemical analysis revealed that the gene product of this enzyme was highly expressed in PCK kidneys and decreased by PIO. These data show that PIO alters the expression of genes involved in cell cycle progression, cell proliferation, and fatty acid metabolism

Mapping the Galactic Halo I. The `Spaghetti' Survey

We describe a major survey of the Milky Way halo designed to test for kinematic substructure caused by destruction of accreted satellites. We use the Washington photometric system to identify halo stars efficiently for spectroscopic followup. Tracers include halo giants (detectable out to more than 100 kpc), blue horizontal branch stars, halo stars near the main sequence turnoff, and the ``blue metal-poor stars'' of Preston et al (1994). We demonstrate the success of our survey by showing spectra of stars we have identified in all these categories, including giants as distant as 75 kpc. We discuss the problem of identifying the most distant halo giants. In particular, extremely metal-poor halo K dwarfs are present in approximately equal numbers to the distant giants for V fainter than 18, and we show that our method will distinguish reliably between these two groups of metal-poor stars. We plan to survey 100 square degrees at high galactic latitude, and expect to increase the numbers of known halo giants, BHB stars and turnoff stars by more than an order of magnitude. In addition to the strong test that this large sample will provide for the question `was the Milky Way halo accreted from satellite galaxies?', we will improve the accuracy of mass measurements of the Milky Way beyond 50 kpc via the kinematics of the many distant giants and BHB stars we will find. We show that one of our first datasets constrains the halo density law over galactocentric radii of 5-20 kpc and z heights of 2-15 kpc. The data support a flattened power-law halo with b/a of 0.6 and exponent -3.0. More complex models with a varying axial ratio may be needed with a larger dataset.Comment: 55 pages, 22 figures, to appear in the Astronomical Journa

Functional Sperm of the Yellowtail (Seriola quinqueradiata) Were Produced in the Small-Bodied Surrogate, Jack Mackerel (Trachurus japonicus).

Production of xenogeneic gametes from large-bodied, commercially important marine species in closely related smaller surrogates with short generation times may enable rapid domestication of the targeted species. In this study we aimed to produce gametes of Japanese yellowtail (Seriola quinqueradiata) using jack mackerel (Trachurus japonicus) as a surrogate with a smaller body size and shorter maturation period. Donor spermatogonia were collected from the testes of yellowtail males and transferred into the peritoneal cavity of 10- and 12-day-old jack mackerel larvae. Twenty days later, 59.5% of the recipients survived of which 88.2% had donor-derived germ cells in their gonads. One year later, genomic DNA templates were prepared from the semen of 96 male recipients and subjected to PCR analyses using primers specific for the yellowtail vasa sequence, resulting in the detection of positive signals in semen from two recipients. The milt collected from the recipients was used for fertilization with yellowtail eggs. Of eight hatchlings obtained from the crosses, two were confirmed to be derived from donor yellowtail by DNA markers, although the others were gynogenetic diploids. These findings indicate that it is possible to produce donor-derived sperm in xenogeneic recipients with smaller body size and shorter generation time by transplanting spermatogonia. Thus, the xenogeneic transplantation of spermatogonia might be a potential tool to produce gametes of large-bodied, commercially important fish although the efficiency of the method requires further improvement. This is the first report demonstrating that donor-derived sperm could be produced in xenogeneic recipient via spermatogonial transplantation in carangid fishes

Combination of Myogenic and Neurogenic Motor Evoked Potential Monitoring During Thoracoabdominal Aortic Surgery

A 64-year-old woman was evaluated for thoracoabdominal aortic aneurysms (TAAAs). Preoperative computed tomography showed a TAAA extending from the level of the diaphragm to the renal arteries. The Adamkiewicz artery (AKA) arose at the Th10 level, close to the aneurysm, and an abdominal aortic prosthesis and left iliac artery aneurysm were detected. Myogenic and neurogenic motor evoked potentials (MEPs) were monitored during the surgical repair of the TAAA, and there were differences between the two types of MEPs during surgery. Both MEPs fell below 50% of their baseline levels during surgery, which suggested critical ischemia, but the decrease in the myogenic MEP occurred at a different time from the decrease in the neurogenic MEP. A time-course analysis suggested that AKA reimplantation was unnecessary and all intercostal arteries were ligated. Both MEPs recovered completely by the end of surgery and there were no postoperative neurologic deficits. Our findings suggest that the combination of myogenic and neurogenic MEP monitoring is helpful in evaluating spinal cord injury during the surgical repair of TAAAs