Purpose: To study a method of the induction of dendritic cells (DCs) from rabbit peripheral blood. Methods: Peripheral
blood cells were removed from rabbit, filtered through nylon mesh. Peripheral blood mononuclear cells (PBMC) were
isolated from the blood cells by Ficoll-Hypaque centrifugation (density of 1.077g/cm3).To obtain DCs, PBMC were
cultured in RPMI1640 medium containing 10% fetal calf serum, 50U/mL penicillin and streptomycin, referred to
subsequently as complete medium, at 37°C in 5% CO2 atmosphere for 4 hours. Nonadherent cells were aspirated,
adherent cells were continued incubated in complete medium, supplemented with granulocyte/macrophage
colony-stimulating factor (GM-CSF, 50ng/ml),and interleukin 4 (IL-4, 50ng/ml) for 9 days. Fluorescein labeled
antibodies(anti-CD14, anti-HLA-DR, anti-CD86) were used to sign cells cultured for 3,6,9 days respectively, Then flow
cytometry was performed. Results: Ratio of anti-HLA-DR and anti-CD86 labeled cells increased with induction time
extension, in contrast with anti-CD14. Conclusion: Dendritic cells can be effectively induced by the method of this
experiment, cell maturation status increased with induction time extension.
Infection of plants with pathogens leads to programmed cell death (PCD) associated with the pathogen-triggered
hypersensitive response (HR) during plant innate immunity. In this study, the effects of infection by virulent
Pseudomonas syringae pv. tomato (Pst) DC3000 and strains harboring avirulence factors AvrRps4 on the induction of
HR-PCD were compared. We used Arabidopsis thaliana plants as materials, which expressed green fluorescent protein
labeled mitochondria (mito-GFP) and green fluorescent protein tagged ATG5 (ATG5-GFP), these GFP are instantaneous
expression. We found both Pst DC3000 and Pst-avrRps4 could induce mitochondria to assemble, the effect of Pst
DC3000 was more obvious. ATG5 was located in chloroplasts after infection with Pst DC3000 or Pst-avrRps4. Under
the condition of Pst-avrRps4, the expression of ATG5 was stronger than Pst DC3000 treatment.
Objective: To observe the differences of cell death and accumulation of reactive oxygen species (ROS) in the process of
infecting Arabidopsis with avirulent Pseudomonas syringae pv. tomato DC3000 (avrB, avrRps4), it will be of great
importance to research the role of plant disease resistance and defense response. Methods: Using WT, AtrbohD and
AtrbohF mutant as experimental materials, we discuss the impact of cell death and ROS on the leaves of Arabidopsis
infected with avirulent Pst DC3000 (avrB, avrRps4), observed by spectral analysis and visualized by DAB and trypan
blue stain. Results: When infected with avirulent Pst DC3000, both WT and AtrbohF mutant line behaved resistance that
exhibited burst of ROS and HR occur, limit senescence and pathogen induced chlorotic cell death. Paradoxically,
AtrbohD mutant behaved susceptible characters that exhibited a small quantity of ROS accumulated and enhanced cell
death. Conclusion: After infection of Arabidopsis with avirulent Pst DC3000, WT exhibited more ROS accumulation
than AtrbohF, and AtrbohD eliminated the majority of total ROS production. Although both WT and AtrbohF mutant
exhibited HR occur, enhanced cell death in AtrbohD mutant.
Autophagy appears to be a highly conserved process from unicellular to multicellular
eukaryotes which contributes to the equilibrium of intracelluar environment. While it would
be harmful to the cells when it is excessive by inducing programmed cell death (PCD). It is a
protein degradation process in which cells recycle cytoplasmic contents when subjected to
environmental stress conditions or during certain stages of development. Previous studies
have demonstrated autophagy can be induced during abiotic or biotic stresses. salicylic acid (SA)
and methyl salicytic (MeSA) are endogenous signal molecules. We found SA and MeSA can
induce autophagy in Arabidopsis thaliana respectively. While autophagy was not induced by SA
or MeSA in tobacco suspension cells under the same concentration and period. The
differences in stuctures or physiological states may contribute to the results.
Wild type Arabidopsis thaliana (ecotype Columbia) and Thellungiella were used as experimental material in this artile.
The leaves of the Arabidopsis and the Thellungiella were treated with different concentrations of NaCl. The
fluorescence emission spectra, the chlorophyll fluorescence and the delayed fluorescence were detected respectively.
We found that there was an obvious change in the photosynthetic efficiency of PS II and the DF intensity of the
Arabidopsis leaves with different concentrations of NaCl treatment. However, there wasnot an obvious change in the
photosynthetic efficiency of PS II and the DF intensity of the Thellungiella leaves with 100mmol/L and 200mmol/L NaCl
treatment. While it also showed that there came to be an obvious stress of the Thellungiella leaves with 300mmol/L
NaCl treatment. The fast chlorophyll fluorescence induction dynamics analysis revealed that the photosynthetic
efficiency of PS II of the Arabidopsis leaves had a sharp decline along with the 100-200 mmol/L NaCl treatment, While
the Thellungiella leaves showed a strong tolerances to salt stress.
Autophagy is a protein degradation process in which cells recycle cytoplasmic contents when subjected to
environmental stress conditions or during certain stages of development. Upon the induction of autophagy, a
double membrane autophagosome forms around cytoplasmic components and delivers them to the vacuole for
degradation. In plants, autophagy has been shown previously to be induced during abiotic stresses including
oxidative stress. Cd, as a toxicity heavy metal, resulted in the production of reactive oxygen species (ROS). In
this paper, we demonstrated that ROS contributed to the induction of autophagy in Cd-stressed Arabidopsis
thaliana. However, pre-incubation with ascorbic acid (AsA, antioxidant molecule) and catalase (CAT, a
H2O2-specific scavenger) decreased the ROS production and the number of autolysosomal-like structures.
Together our results indicated that the oxidative condition was essential for autophagy, as treatment with AsA and
CAT abolished the formation of autophagosomes, and ROS may function as signal molecules to induce autophagy
in abiotic stress.
Studies the superparamagnetic iron oxide (SPIO) label ECV304 method and test its effect on cell physiological
activity .We use different concentration PLL-SPIO incubate ECV304 cells, Prussian blue dye was taken to examine
SPIO mark efficiency, DCF dyeing and PI & Annexin the VFITC double dyeing was carried out to analyze cell
apoptosis. The result shows that 25μg/ml gave the most efficient labeling concentration, there is also no toxic effect on
cells in this concentration. This outcome can be used in MRI of living body cell transplant tracking.
Propose: To study the optimal concentration and time of incubation of human lung adenocarcinoma cell line (SPC-A-1)
labeled with superparamagnetic iron oxide (SPIO) particles in vitro. Methods: Human lung adenocarcinoma cell line
(SPC-A-1) was cultured with different concenration of SPIO and different time of incubation (labeled with media
containing Fe-PLL: 25μg /mL, 100μg /mL, and 200 μg /mL, and for 30min, 90min, 180min. The phagocytosis of the
cells was observed by laser scanning confocal microscopy (LSCM) to determine particle uptake and their distribution in
cells. Results: Human lung adenocarcinoma cells(SPC-A-1) have taken up a large amount of SPIO particles within the
first 3h. Conclusion: In this study, the concentration of iron with 25μg/ml SPIO and time of incubation for 30min is the
optimal condition for labeling the SPC-A-1 with SPIO.
Changes of chlorophyll fluorescence emission spectra of different position leaves were investigated during the later
growth stages of rice leaves. Results showed that natural illumination induced a shift change of fluorescence emission
intensity (both in P685 and P735) during the progressive senescence of rice leaves. These data suggested that changes of
chlorophyll fluorescence emission spectra of different position leaves were possible due to the declined of energy and
electron transport activities of PSII reaction center. On the other hand, different position leaves of rice were treated with
3-(3,4 dichlorophenyl)-1,1-dimethyl-urea (DCMU), the results showed that DCMU had a significant effect on Chl
fluorescence emission spectra (both in P685 and P735) under room temperature and different position leaves had a
different sensibility of DCMU. It is concluded that natural illumination plays an important role in the progressive
senescence process of rice leaves.
The volatile halogenated methyl ethyl ether is used as anesthetic to inhibit actin-based dynamics directly or indirectly in
animal cells. In plant cells, most intracellular movements are related with actin pathways too. We utilized isoflurane to
study the dynamic choloroplast organization in unicellular baby and adult alga Acetabularia acetabulum. Fluorescence
spectra were measured and choloroplast movements were recorded by confocal laser scanning microscope (CLSM)
imaging in in Acetabularia acetabulum. Isoflurane was effective in the unicellular baby and adult organisms and showed
time- dependent actin-inhibition patterns. Acetabularia acetabulum cells were treated for different times with
isoflurane saturated solutions in artificial seawater (it was defined to be 100% isoflurane). The intensity of fluorescence
at 680nm and 730nm were progressively decreased at 100% isoflurane. It was remarkable difference between
fluorescence spectra of baby and adult Acetabularia were inhibited by isoflurance, adult Acetabularia cells showed more
sensitive. Whereas the choloroplast in Acetabularia acetabulum was commendably imaged by CLSM at 20 and 40 zoom.
Employing a luminescence spectrometer the usefulness of light-induced delayed luminescence (DL) for the
detection of aflatoxinB1 (AfB1) contamination in whole rice was studied. Rice was artificially contaminated with
Aspergillus flavus (7.5×105 conidia/ml) and incubated for 0 - 72 h. The DL spectra of contaminated rice were
obtained by a luminescence spectrometer. The correlation between the spectra and contamination levels was
established. The DL intensity has a negative correlation with the AfB1 concentration. The results suggested that the
DL technique might be useful for the rapid and non-invasive evaluation of AfB1 contamination levels in whole rice.
Cerebrovascular disease is one of the leading causes of death, and approximately 50% of survivors have a residual
neurologic deficit and greater than 25% require chronic care. Cerebrovascular reserve capacity (CVRC) describes how
far cerebral perfusion can increase from a baseline value after stimulation. High blood pressure is the most important
independent risk factor for stroke and other vascular diseases. The incidence of stroke in the hypertensive is six times
higher than in the patient with normal blood pressure. CVRC in the hypertensive was even lower than in control patients.
MR perfusion weighted imaging (MR PWI) with the well-established acetazolamide (ACZ) stimulation test has been
used for assessing brain function. The aim of this work is to assess the cerebrovascular reserve capacity by MR PWI with
"ACZ" tolerance test in spontaneous hypertensive rat (SHR) and to identify its value in evaluating the CVRC.
Experimental animal including 3 groups: Wistar-Kyoto rats (WKY) (12-week-old) as control group, SHR (12-week-old
and 20-week-old) as experimental group. MR PWI was performed respectively before and after acetazolamide
administrated orally in 3 groups on a clinical 1.5 Tesla GE Signa MR fx/i whole-body MR system. The ROI was chosen
in the bilateral frontal lobe to measure the value of rCBV, rCBF and MTT. The results showed that before ACZ-test,
there was statistic differences between the WKY and SHR(12-week-old), and between SHR(12-week-old) and
SHR(20-week-old) in the values of rCBV and rCBF (P>0.05), and after ACZ-test, there were statistic differences
between WKY and SHR (20-week-old), and between SHR(12-week-old) and SHR(20-week-old) in the rCBV value (P<0.05). It is concluded that the method of MRI PWI combined with the "ACZ stress test" can provide more qualitative
and half-quantitative information on the cerebral perfusion to evaluate the CVRC in SHR.
Gerbera hybrida (Shenzhen No.5) seedlings' inflorescence development was divided into six stages (P1-P6). With
these six stages petal, delayed luminescence (DL) were observed during petal development using lab-made detector
system, fluorescence spectrum and confocal imaging were also observed. The results showed that the intensity of DL
were increased during P1-P4 and decreased in P5 and P6; with the excitation wavelength of 488 nm, fluorescence
spectra were obviously different during P1-P6 stages; imaging of chlorophyll autofluorescence by confocal laser
scanning microscopy (CLSM) showed that the intensity were stronger in P3 than in P1, while P6 stage autofluorescence
only displayed in guard cell of epidermis. Our results suggested that the DL technique, combining with fluorescence
spectra and CLSM imaging, might be useful for the rapid and noninvasive evaluation of chlorophyll content and
degradation in petal development in Gerbera hybrida.
Ultraweak bio-chemiluminescence (UBC) from germinating soybean (Glycine max L. Merr) cotyledon under mechanical wounding was observed by a high sensitivity imaging system based on an intensified charge couple device (ICCD) detector or a high sensitive single photon counter (SPC) device. The UBC imaging showed that the intensity of UBC at the injury location on a wounded green cotyledon was greater than on a wounded etiolated cotyledon. Based on results with a SPC UBC intensity of wounded green cotyledon was high at first and then gradually decreased. The emission spectrum of wounded green cotyledon had a greater proportion of red light. The increase in UBC of wounded etiolated soybean cotyledon was less than that of green ones. The emission spectrum of wounded etiolated cotyledon had a greater amount of orange light. The data suggest that most of the UBC in green cotyledon was due to damaged chloroplasts and mitochondria. Our data suggest that oxidation of damage's tissue lead to the production of ROS. Electronic excitation energy was transfered from the excited molecules by ROS to Chl-a in the thylakoid membranes, so
the intensity of UBC in the wounded soybean green cotyledon was obviously higher than in the wounded etiolated soybean cotyledon.
KEYWORDS: Luminescence, Chemiluminescence, Fluorescence spectroscopy, Oxygen, Chemical analysis, Life sciences, Biomedical optics, Single photon, Data modeling, Industrial chemicals
BSA (Bovine Serum Albumin) can enlarge the CL intensity of FCLA(3,7-dihydro-6-{4-{2-(N'-(5-fluoresceinyl) thioureido)ethoxy}phenyl}-2-methylimi-dazo{1,2-a}pyrazin-3-one dosium salt) to 763%. This report presents novel methods for determination of Aflatoxin B1 (AfB1) mediated by FCLA+BSA. The concentration of AFB1 showed an obvious positive correlation with the chemiluminescence (CL) intensity mediated by FCLA+BSA, correlative coefficient R@0.94. This method could measure accurately ng/ml of AfB1 concentration. 365nm as excitated wavelength, 440nm and 520nm-two fluorescence peaks of FCLA+BSA+AfB1 were found. The fluorescence intensity of peak at 440nm showed an obvious positive correlation with the concentration of AFB1, R@0.97; the fluorescence intensity of peak at 520nm showed a positive correlation with the concentration of AFB1, R@0.90. Comparing the peak of FCLA, FCLA+BSA and FCLA+BSA+AfB1 had a 6nm Einstein shift (red shift). The study suggested that CL and fluorescence spectrum methods mediated by FCLA+BSA might be applicable to the determination of AfB1 concentration.
As a chemiluminescence (CL) probe, 3,7-dihydro-6-{4-{2-(N’-(5-fluoresceinyl) thioureido)ethoxy}phenyl}-2-met
-hylimi-dazo{1,2-a}pyrazin-3-one dosium salt (FCLA) can sensitively and specifically react with singlet oxygen (1O2 ) and superoxide(O2""). BSA (Bovine Serum Albumin) can enlarge the CL intensity of FCLA to 860%. This report presents a novel method for determination of Aflatoxin B1 (AfB1) mediated by FCLA+BSA. The concentration of AFB1 showed an obvious positive correlation with the CL intensity mediated by FCLA+BSA. This method could measure accurately ng/ml of AfB1 concentration. At the same time, the fluorescence spectrum of FCLA+BSA and FCLA+BSA+AfB1 were measured respectively, which showed that the fluorescence intensity of FCLA+BSA+AfB1 was higher than FCLA+BSA. Comparing the peak value of FCLA, FCLA+BSA and FCLA+BSA+AfB1 had a 6nm Einstein shift (red shift). The study suggested that CL method mediated by FCLA+BSA might be applicable to the determination of AfB1 concentration.
With high-sensitivity single-photon counter, spontaneous chemiluminescence (CL) spectrum and different temperature study on rice (Oryza sativa L.) seeds during early imbibition were carried out. The emission spectrum of whole rice seed, rice and coat had a greater proportion of red light during early imbibition. Comparing with spontaneous CL of barley (Hordeum vulgare L.) and soybean (Glycine max L. Merr) seeds, the spontaneous CL of rice seeds had a nonlinear, logarithmic-like increase of intensity in the T range 30-50°C, the Van't Hoff coefficient Q10=IT+10/IT is equal to 2, which led us to the conclusion that spontaneous CL of rice seed during early imbibition partly came from enzyme catalyzing chemistry reaction.
Ultra-weak bio-chemiluminescence (UBC) from germinating soybean (Glycine max L. Merr) cotyledon under mechanical wounding was observed by a high sensitive single photon counter (SPC) device. SPC measuring results showed that the UBC intensity of wounding green cotyledon was very high firstly and reached to a stationary state after about 5 minutes. Wounding-induced emission could be suppressed by wounding treatment with sodium azide. While the intensity increase of the wounded etiolated soybean cotyledon was less than the green one. Deutenum oxide both amplified the emission intensity of green and etiolated. Singlet oxygen (102) is a main cause of the emission during the wounding phase. We suggested that the main process was probably that chioroplast and mitochondria damage at the wounded place and leaded to ROS generation, such as H202, 102 etc. The electronic excitation energy transfered from the excited molecules by ROS to Chl-a in the thylakoid membranes by way of relatively high quantum yields, so the intensity of UBC in the wounded soybean green cotyledon was obviously higher than in the wounded etiolated soybean cotyledon.
Ultraweak Chemiluminescence (UCL) studies of different aging degree of rice (Oryza sativa L.) seeds stored in a high temperature 40 degree(s)C and high relative humidity 90% environment (0 day, 8 days, 15 days, and 22 days) were carried out. We firstly observed that aging degree of rice seeds was positive correlation with ultraweak chemiluminescence during the early imbibition (0-1h). Addition of water to rice seeds stimulates ultraweak chemiluminescence, the intensity of which depends upon aging degree of seeds. The shorter the seed accelerated aging time was, the higher the intensity of the UCL in the early imbibition period, the lower hydrogen peroxide (H2O2) concentration of rice seeds, the higher percentage seed germination. The germination and superoxide dismutase (SOD) activity of dry rice seeds was obvious positive correlation with the intensity of UCL. While catalase (CAT) activity of rice seeds was determined. Mechanism of ultraweak chemiluminescence was discussed. It was concluded that the store time of rice seeds could be judged from their UCL characters during the early imbibition period, which might be a way to examine vigor of seeds.
Access to the requested content is limited to institutions that have purchased or subscribe to SPIE eBooks.
You are receiving this notice because your organization may not have SPIE eBooks access.*
*Shibboleth/Open Athens users─please
sign in
to access your institution's subscriptions.
To obtain this item, you may purchase the complete book in print or electronic format on
SPIE.org.
INSTITUTIONAL Select your institution to access the SPIE Digital Library.
PERSONAL Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.