To clarify the physiologic roles of granulocyte colony-stimulating factor (G-CSF) in infectious states in vivo, we examined the serum levels of G-CSF in patients with infection. Serum samples from 24 patients in the acute stage of infection (14 men and 10 women, age 65 to 101, without hematologic disorders), as well as samples from 32 age- matched normal elderly volunteers were investigated. Sixteen of the initial 24 patients were reexamined after the recovery phase. G-CSF levels were examined by quantitative enzyme immunoassay. The G-CSF level in normal elderly controls, 25.3 +/- 19.7 pg/mL, was not different from that reported in other findings. There was no statistically significant relationship between their G-CSF level and peripheral white blood cell count or neutrophilic granulocyte count. The G-CSF level in the acute stage of infection was 731.8 +/- 895.0 pg/mL, with a range of 30 to 3,199 pg/mL. There was no significant difference in G-CSF levels between patients with respiratory tract infection and those with urinary tract infection. In all 16 cases examined, the serum G-CSF level in the acute stage of infection was significantly higher than that after recovery phase, the latter being the same as the level in normal elderly controls. G-CSF must therefore play a significant role in human infectious states in vivo.
This study was designed to detect apoptosis in the human amnion and to elucidate the signalling pathway involved in its regulation. Samples of human amnion were obtained from 34 women (weeks 11-42 of gestation) and studied using the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labelling (TUNEL) method with light microscopy (LM) and transmission electron microscopy (TEM). Apoptotic regulators in the samples were studied by immunohistochemistry and caspase activity assay. The TUNEL method with LM demonstrated that the percentage of TUNEL-positive cells in the amniotic epithelium was the highest in weeks 40-41 of gestation (P < 0.05) independent of the onset of labour, and the cells were often detached from the epithelium into the amniotic cavity at term. The TUNEL method with TEM clearly showed the characteristic features of apoptosis such as the nuclear condensed chromatin with abundant free 3'-OH DNA ends, cell shrinkage and a decrease in the number of desmosomes, except for the presence of apoptotic bodies. Fas and Fas ligand (FasL) were constantly expressed on apical membranes of amniotic epithelial cells from weeks 16-27 through to 40-41 of gestation, while no Bcl-2 expression was observed throughout the gestational periods. Activities of caspase-3 and caspase-8, but not of caspase-9, were higher in weeks 40-41 than those from weeks 16-27 of gestation (P < 0.01). We conclude that apoptosis in term amniotic epithelium is independent of Bcl-2 regulation and onset of labour, and may play an important role in the fragility and rupture of human fetal membranes at term.
A simple and improved dwarf rice (Oryza sativa var Tan-ginbozu) lamina inclination bioassay for brassinosteroids (BRs) was developed based on a previously published method (K Takeno, RP Pharis [1982] Plant Cell Physiol 23: 1275-1281). The assay used 3-day-old intact seedlings, and detection of BR was made more sensitive by synergizing the response to BR with indole-3-acetic acid (IAA). The minimum detectable amount of BR was less than 0.1 nanogram/rice plant (brassinolide equivalents). Purification steps for isolation of BR from tissue scrapings taken from the cambial region of Scots pine (Pinus silverstris) harvested during the period of rapid wood production were guided by this assay. After column chromatography (silica gel and PrepPak C18) and reversed phase Cie high performance liquid chromatography, the biologically active fractions were analyzed by gas chromatography-mass spectrometry (GC-MS) and/or GC-MS-selected ion monitoring. Two BRs, castasterone (major) and brassinolide (minor) were identified. This is the first identification of BR from the cambial region of a conifer.Brassinosteroids (BRs)4 are steroidal hormones that promote plant growth. To date, 28 naturally occurring BRs have been identified in various plant tissues (2,4,5,8,16), ranging from higher plants to a green alga ( 15). In conifers, castasterone (scheme 1, structure 2) and typhasterol (2-deoxycastasterone (scheme 1, structure 3) have been identified in pine pollen
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