14 research outputs found
Effect of neonatal hypoxia on the development of intraspinal serotonergic fibers in relation to spinal motoneurons
http://dx.doi.org/10.1016/j.braindev.2008.12.009Serotonin (5-hydroxytryptamine; 5-HT)-containing neurons trophically affect target neurons and modulate central nervous system neuronal activity. We studied effects of neonatal hypoxia on postnatal development of intraspinal 5-HT fibers in spinal motoneuron pools. Postnatal day (PND) 0 Sprague–Dawley rats received a hypoxic load and survivors were used for histological analyzes on PNDs 1, 7, and 14. Spinal motoneurons were labeled using choleratoxin B subunit as a retrograde neurotracer, and 5-HT fibers were detected immunohistochemically. On PND 1, 5-HT fibers were present in the lateral portion of the ventral horn at the cervical level, but were sparsely distributed at the lumbar level. On PND 14, cervical and lumbar level distributions were nearly identical. The 5-HT fibers and varicosities in close apposition to motoneurons increased from PNDs 1–14, however, the close apposition of cervical motoneurons was significantly different from lumbar motoneurons only on PND 1. Density of 5-HT fibers in control and hypoxic rats was not different on PND 1, while those in hypoxic rats were significantly reduced on PND 14. Close appositions of lumbar motoneurons were reduced more than cervical MNs after neonatal hypoxia. Neurodevelopmental deficit after neonatal hypoxia with a rostro-caudal gradient is associated with significant changes in the 5-HT system
Sensitive Multiplexed Quantitative Analysis of Autoantibodies to Cancer Antigens with Chemically <i>S-</i>Cationized Full-Length and Water-Soluble Denatured Proteins
Humoral
immune responses against tumor-associated antigens (TAAs)
or cancer/testis antigens (CTAs) aberrantly expressed in tumor cells
are frequently observed in cancer patients. Recent clinical studies
have elucidated that anticancer immune responses with increased levels
of anti-TAA/CTA antibodies
improve cancer survival rates. Thus, these antibody levels are promising
biomarkers for diagnosing the efficiency of cancer immunotherapy.
Full-length antigens are favored for detecting anti-TAA/CTA antibodies
because candidate antigen proteins contain multiple epitopes throughout
their structures. In this study, we developed a methodology to prepare
purified water-soluble and full-length antigens by using cysteine
sulfhydryl group cationization (<i>S-</i>cationization)
chemistry. <i>S</i>-Cationized antigens can be prepared
from bacterial inclusion bodies, and they exhibit improved protein
solubility but preserved antigenicity. Anti-TAA/CTA antibodies detected
in cancer patients appeared to recognize linear epitopes, as well
as conformational epitopes, and because the frequency of cysteine
side-residues on the epitope–paratope interface was low, any
adverse effects of <i>S-</i>cationization were virtually
negligible for antibody binding. Furthermore, <i>S-</i>cationized
antigen-immobilized Luminex beads could be successfully used in highly
sensitive quantitative-multiplexed assays. Indeed, patients with a
more broadly induced serum anti-TAA/CTA antibody level showed improved
progression-free survival after immunotherapy. The comprehensive anti-TAA/CTA
assay system, which uses <i>S-</i>cationized full-length
and water-soluble recombinant antigens, may be a useful diagnostic
tool for assessing the efficiency of cancer immunotherapy