Abstract: We develop new nonlinear methods of
immunofluorescence analysis for a sensitive technology of
respiratory burst reaction of DNA fluorescence due to oxidative
activity in the peripheral blood neutrophils. Histograms in flow
cytometry experiments represent a fluorescence flashes frequency as
functions of fluorescence intensity. We used the Shannon-Weaver
index for definition of neutrophils- biodiversity and Hurst index for
definition of fractal-s correlations in immunofluorescence for
different donors, as the basic quantitative criteria for medical
diagnostics of health status. We analyze frequencies of flashes,
information, Shannon entropies and their fractals in
immunofluorescence networks due to reduction of histogram range.
We found the number of simplest universal correlations for
biodiversity, information and Hurst index in diagnostics and
classification of pathologies for wide spectra of diseases. In addition
is determined the clear criterion of a common immunity and human
health status in a form of yes/no answers type. These answers based
on peculiarities of information in immunofluorescence networks and
biodiversity of neutrophils. Experimental data analysis has shown the
existence of homeostasis for information entropy in oxidative activity
of DNA in neutrophil nuclei for all donors.
Abstract: Autofluorescence (AF) bronchoscopy is an
established method to detect dysplasia and carcinoma in situ (CIS).
For this reason the “Sotiria" Hospital uses the Karl Storz D-light
system. However, in early tumor stages the visualization is not that
obvious. With the help of a PC, we analyzed the color images we
captured by developing certain tools in Matlab®. We used statistical
methods based on texture analysis, signal processing methods based
on Gabor models and conversion algorithms between devicedependent
color spaces. Our belief is that we reduced the error made
by the naked eye. The tools we implemented improve the quality of
patients' life.
Abstract: Acute kidney injury (AKI) is a new worldwide public
health problem. A diagnosis of this disease using creatinine is still a
problem in clinical practice. Therefore, a measurement of biomarkers
responsible for AKI has received much attention in the past couple
years. Cytokine interleukin-18 (IL-18) was reported as one of the
early biomarkers for AKI. The most commonly used method to
detect this biomarker is an immunoassay. This study used a planar
platform to perform an immunoassay using fluorescence for
detection. In this study, anti-IL-18 antibody was immobilized onto a
microscope slide using a covalent binding method. Make-up samples
were diluted at the concentration between 10 to 1000 pg/ml to create
a calibration curve. The precision of the system was determined
using a coefficient of variability (CV), which was found to be less
than 10%. The performance of this immunoassay system was
compared with the measurement from ELISA.
Abstract: Ethanol is generally used as a therapeutic reagent against Hepatocellular carcinoma (HCC or hepatoma) worldwide, as it can induce Hepatocellular carcinoma cell apoptosis at low concentration through a multifactorial process regulated by several unknown proteins. This paper provides a simple and available proteomic strategy for exploring differentially expressed proteins in the apoptotic pathway. The appropriate concentrations of ethanol required to induce HepG2 cell apoptosis were first assessed by MTT assay, Gisma and fluorescence staining. Next, the central proteins involved in the apoptosis pathway processs were determined using 2D-PAGE, SDS-PAGE, and bio-software analysis. Finally the downregulation of two proteins, AFP and survivin, were determined by immunocytochemistry and reverse transcriptase PCR (RT-PCR) technology. The simple, useful method demonstrated here provides a new approach to proteomic analysis in key bio-regulating process including proliferation, differentiation, apoptosis, immunity and metastasis.
Abstract: Identifying the nature of protein-nanoparticle
interactions and favored binding sites is an important issue in
functional characterization of biomolecules and their physiological
responses. Herein, interaction of silver nanoparticles with lysozyme
as a model protein has been monitored via fluorescence spectroscopy.
Formation of complex between the biomolecule and silver
nanoparticles (AgNPs) induced a steady state reduction in the
fluorescence intensity of protein at different concentrations of
nanoparticles. Tryptophan fluorescence quenching spectra suggested
that silver nanoparticles act as a foreign quencher, approaching the
protein via this residue. Analysis of the Stern-Volmer plot showed
quenching constant of 3.73 μM−1. Moreover, a single binding site in
lysozyme is suggested to play role during interaction with AgNPs,
having low affinity of binding compared to gold nanoparticles.
Unfolding studies of lysozyme showed that complex of lysozyme-
AgNPs has not undergone structural perturbations compared to the
bare protein. Results of this effort will pave the way for utilization of
sensitive spectroscopic techniques for rational design of
nanobiomaterials in biomedical applications.
Abstract: Three sulphonic acid-doped polyanilines were
synthesized through chemical oxidation at low temperature (0-5 oC)
and potential of these polymers as sensing agent for O2 gas detection
in terms of fluorescence quenching was studied. Sulphuric acid,
dodecylbenzene sulphonic acid (DBSA) and camphor sulphonic acid
(CSA) were used as doping agents. All polymers obtained were dark
green powder. Polymers obtained were characterized by Fourier
transform infrared spectroscopy, ultraviolet-visible absorption
spectroscopy, thermogravimetry analysis, elemental analysis,
differential scanning calorimeter and gel permeation
chromatography. Characterizations carried out showed that polymers
were successfully synthesized with mass recovery for sulphuric aciddoped
polyaniline (SPAN), DBSA-doped polyaniline (DBSA-doped
PANI) and CSA-doped polyaniline (CSA-doped PANI) of 71.40%,
75.00% and 39.96%, respectively. Doping level of SPAN, DBSAdoped
PANI and CSA-doped PANI were 32.86%, 33.13% and
53.96%, respectively as determined based on elemental analysis.
Sensing test was carried out on polymer sample in the form of
solution and film by using fluorescence spectrophotometer. Samples
of polymer solution and polymer film showed positive response
towards O2 exposure. All polymer solutions and films were fully
regenerated by using N2 gas within 1 hour period. Photostability
study showed that all samples of polymer solutions and films were
stable towards light when continuously exposed to xenon lamp for 9
hours. The relative standard deviation (RSD) values for SPAN
solution, DBSA-doped PANI solution and CSA-doped PANI
solution for repeatability were 0.23%, 0.64% and 0.76%,
respectively. Meanwhile RSD values for reproducibility were 2.36%,
6.98% and 1.27%, respectively. Results for SPAN film, DBSAdoped
PANI film and CSA-doped PANI film showed the same
pattern with RSD values for repeatability of 0.52%, 4.05% and
0.90%, respectively. Meanwhile RSD values for reproducibility were
2.91%, 10.05% and 7.42%, respectively. The study on effect of the
flow rate on response time was carried out using 3 different rates
which were 0.25 mL/s, 1.00 mL/s and 2.00 mL/s. Results obtained
showed that the higher the flow rate, the shorter the response time.
Abstract: Lanthanide-doped upconversion nanoparticles which can convert near-infrared lights to visible lights have attracted growing interest because of their great potentials in fluorescence imaging. Upconversion fluorescence imaging technique with excitation in the near-infrared (NIR) region has been used for imaging of biological cells and tissues. However, improving the detection sensitivity and decreasing the absorption and scattering in biological tissues are as yet unresolved problems. In this present study, a novel NIR-reflected multispectral imaging system was developed for upconversion fluorescent imaging in small animals. Based on this system, we have obtained the high contrast images without the autofluorescence when biocompatible UCPs were injected near the body surface or deeply into the tissue. Furthermore, we have extracted respective spectra of the upconversion fluorescence and relatively quantify the fluorescence intensity with the multispectral analysis. To our knowledge, this is the first time to analyze and quantify the upconversion fluorescence in the small animal imaging.
Abstract: Utilization of bagasse ash for silica sources is one of
the most common application for agricultural wastes and valuable
biomass byproducts in sugar milling. The high percentage silica
content from bagasse ash was used as silica source for sodium
silicate solution. Different heating temperature, time and acid
treatment were studies for silica extraction. The silica was
characterized using various techniques including X-ray fluorescence,
X-ray diffraction, Scanning electron microscopy, and Fourier
Transform Infrared Spectroscopy method,. The synthesis conditions
were optimized to obtain the bagasse ash with the maximum silica
content. The silica content of 91.57 percent was achieved from
heating of bagasse ash at 600°C for 3 hours under oxygen feeding
and HCl treatment. The result can be used as value added for bagasse
ash utilization and minimize the environmental impact of disposal
problems.
Abstract: Breast carcinoma is the most common form of cancer
in women. Multicolour fluorescent in-situ hybridisation (m-FISH) is
a common method for staging breast carcinoma. The interpretation
of m-FISH images is complicated due to two effects: (i) Spectral
overlap in the emission spectra of fluorochrome marked DNA probes
and (ii) tissue autofluorescence. In this paper hyper-spectral images of
m-FISH samples are used and spectral unmixing is applied to produce
false colour images with higher contrast and better information
content than standard RGB images. The spectral unmixing is realised
by combinations of: Orthogonal Projection Analysis (OPA), Alterating
Least Squares (ALS), Simple-to-use Interactive Self-Modeling
Mixture Analysis (SIMPLISMA) and VARIMAX. These are applied
on the data to reduce tissue autofluorescence and resolve the spectral
overlap in the emission spectra. The results show that spectral unmixing
methods reduce the intensity caused by tissue autofluorescence by
up to 78% and enhance image contrast by algorithmically reducing
the overlap of the emission spectra.
Abstract: Low silica type X (LSX) Zeolite is one of useful
material in many manufacturing due to the advantage properties
including high surface area, stability, microporous crystalline
aluminosilicates and positive ion in an extra–framework. The LSX
was used rice husk silica source which obtained by leaching with
hydrochloric acid and calcination at 500 C. To improve the
synthesis method, the LSX was crystallizated in Teflon–lined
autoclave will expedite deceasing of the amorphous particles. The
mixed gel with composition of 5.5 Na2O : 1.65 K2O : Al2O3 : 2.2
SiO2 : 122 H2O was crystallized in different container
(Polypropylene bottom and Teflon–lined autoclave). The obtained
powder was characterized by X–ray diffraction (XRD), X–ray
fluorescence spectrometry, N2 adsorption-desorption analysis BET
surface area Scanning electron microscopy (SEM) and Fourier
transform infrared spectroscopy to justify the quality of zeolite. The
results showed the crystallized zeolite in Teflon lined autoclave has
102.8 nm of crystal size, 286 m2/g of surface area and fewer amounts
of round amorphous particles when compared with the crystallized
zeolite in Polypropylene.
Abstract: Absorption and fluorescence spectra of quinine
sulphate (QSD) have been recorded at room temperature in wide
range of solvents of different polarities. The ground-state dipole
moment of QSD was obtained from quantum mechanical calculations
and the excited state dipole moment of QSD was estimated from
Bakhshiev-s and Kawski-Chamma-Viallet-s equations by means of
solvatochromic shift method. Higher value of dipole moment is
observed for excited state as compared to the corresponding ground
state value and this is attributed to the more polar excited state of
QSD.
Abstract: To investigate effect of salt stress on Chlorophyll
fluorescence four cultivars (fong,star,chamran and kharchia) of wheat
(Triticum aestivum) plants subjected to salinity levels ( control,8,12
and 16 dsm-1 ) from one week after emergence to the end of stem
elongation under greenhouse condition . results showed that quantum
yield of photosystem II from light adopted leaves (ΦPSII),
Photochemical quenching (qP) ,quantum yield of dark adopted leaves
(fv/fm) and non photochemical quenching (NPq) were affected by
salt stress . Salinity levels affected photosynthetic rate. Star and fong
cultivars showed minimum and maximum levels of photosynthetic
rate in respectively. Minimum photosynthetic rate differences
between levels of salinity were shown in Kharchia. Shoot dry matter
of all cultivars decreased by increasing salinity levels. Results
showed that non photochemical quenching by salinity levels attribute
to the decreases in shoot dry matter.
Abstract: The purpose of this study is to present a non invasive
method for the marginal adaptation evaluation in class V composite
restorations. Standardized class V cavities, prepared in human
extracted teeth, were filled with Premise (Kerr) composite. The
specimens were thermo cycled. The interfaces were examined by
Optical Coherence Tomography method (OCT) combined with the
confocal microscopy and fluorescence. The optical configuration
uses two single mode directional couplers with a superluminiscent
diode as the source at 1300 nm. The scanning procedure is similar to
that used in any confocal microscope, where the fast scanning is enface
(line rate) and the depth scanning is much slower (at the frame
rate). Gaps at the interfaces as well as inside the composite resin
materials were identified. OCT has numerous advantages which
justify its use in vivo as well as in vitro in comparison with
conventional techniques.
Abstract: To evaluate genetic variation of wheat (Triticum
aestivum) affected by heat and drought stress on eight Australian
wheat genotypes that are parents of Doubled Haploid (HD) mapping
populations at the vegetative stage, the water stress experiment was
conducted at 65% field capacity in growth room. Heat stress
experiment was conducted in the research field under irrigation over
summer. Result show that water stress decreased dry shoot weight
and RWC but increased osmolarity and means of Fv/Fm values in all
varieties except for Krichauff. Krichauff and Kukri had the
maximum RWC under drought stress. Trident variety was shown
maximum WUE, osmolarity (610 mM/Kg), dry mater, quantum yield
and Fv/Fm 0.815 under water stress condition. However, the
recovery of quantum yield was apparent between 4 to 7 days after
stress in all varieties. Nevertheless, increase in water stress after that
lead to strong decrease in quantum yield. There was a genetic
variation for leaf pigments content among varieties under heat stress.
Heat stress decreased significantly the total chlorophyll content that
measured by SPAD. Krichauff had maximum value of Anthocyanin
content (2.978 A/g FW), chlorophyll a+b (2.001 mg/g FW) and
chlorophyll a (1.502 mg/g FW). Maximum value of chlorophyll b
(0.515 mg/g FW) and Carotenoids (0.234 mg/g FW) content
belonged to Kukri. The quantum yield of all varieties decreased
significantly, when the weather temperature increased from 28 ÔùªC to
36 ÔùªC during the 6 days. However, the recovery of quantum yield
was apparent after 8th day in all varieties. The maximum decrease
and recovery in quantum yield was observed in Krichauff. Drought
and heat tolerant and moderately tolerant wheat genotypes were
included Trident, Krichauff, Kukri and RAC875. Molineux, Berkut
and Excalibur were clustered into most sensitive and moderately
sensitive genotypes. Finally, the results show that there was a
significantly genetic variation among the eight varieties that were
studied under heat and water stress.
Abstract: The C3 plants are frequently suffering from exposure
to high temperature stress which limits the growth and yield of these
plants. This study seeks to clarify the physiological mechanisms of
heat tolerance in relation to oxidative stress in C3 species. Fifteen C3
species were exposed to prolonged moderately high temperature
stress 36/30°C for 40 days in a growth chamber. Chlorophyll
fluorescence (Fv/Fm) showed great difference among species at 40
days of the stress. The species showed decreases in Fv/Fm and
increases in malondialdehyde (MDA) content under stress condition
as well as negative correlation between Fv/Fm and MDA (r = -0.61*)
at 40 days of the stress. Hydrogen peroxide (H2O2) content before
and after stress in addition to its response under stress showed great
differences among species. The results suggest that the difference in
heat tolerance among C3 species is closely associated with the ability
to suppress oxidative damage but not with the content of reactive
oxygen species (ROS) which is regulated by complex network.
Abstract: pH-sensitive drug targeting using nanoparticles for
cancer chemotherapy have been spotlighted in recent decades. Graft
copolymer composed of poly (L-histidine) (PHS) and dextran
(DexPHS) was synthesized and pH-sensitive nanoparticles were
fabricated for pH-responsive drug delivery of doxorubicin (DOX).
Nanoparticles of DexPHS showed pH-sensitive changes in particle
sizes and drug release behavior, i.e. particle sizes and drug release rate
were increased at acidic pH, indicating that DexPHS nanoparticles
have pH-sensitive drug delivery potentials. Antitumor activity of
DOX-incorporated DexPHS nanoparticles were studied using CT26
colorectal carcinoma cells. Results indicated that fluorescence
intensity was higher at acidic pH than basic pH. These results
indicated that DexPHS nanoparticles have pH-responsive drug
targeting.
Abstract: Bonding has become a routine procedure in several
dental specialties – from prosthodontics to conservative dentistry and
even orthodontics. In many of these fields it is important to be able to
investigate the bonded interfaces to assess their quality. All currently
employed investigative methods are invasive, meaning that samples
are destroyed in the testing procedure and cannot be used again. We
have investigated the interface between human enamel and bonded
ceramic brackets non-invasively, introducing a combination of new
investigative methods – optical coherence tomography (OCT),
fluorescence OCT and confocal microscopy (CM). Brackets were
conventionally bonded on conditioned buccal surfaces of teeth. The
bonding was assessed using these methods. Three dimensional
reconstructions of the detected material defects were developed using
manual and semi-automatic segmentation. The results clearly prove
that OCT, fluorescence OCT and CM are useful in orthodontic
bonding investigations.
Abstract: Experimental investigations were carried out in the
Manchester Tidal flow Facility (MTF) to study the flow patterns in
the region around and adjacent to a hypothetical headland in tidal
(oscillatory) ambient flow. The Planar laser-induced fluorescence
(PLIF) technique was used for visualization, with fluorescent dye
released at specific points around the headland perimeter and in its
adjacent recirculation zone. The flow patterns can be generalized into
the acceleration, stable flow and deceleration stages for each halfcycle,
with small variations according to location, which are more
distinct for low Keulegan-Carpenter number (KC) cases. Flow
patterns in the mixing region are unstable and complex, especially in
the recirculation zone. The flow patterns are in agreement with
previous visualizations, and support previous results in steady
ambient flow. It is suggested that the headland lee could be a viable
location for siting of pollutant outfalls.
Abstract: In this study we investigate silica nanoparticle (SiO2- NP) effects on the structure and phase properties of supported lipid monolayers and bilayers, coupling surface pressure measurements, fluorescence microscopy and atomic force microscopy. SiO2-NPs typically in size range of 10nm to 100 nm in diameter are tested. Our results suggest first that lipid molecules organization depends to their nature. Secondly, lipid molecules in the vinicity of big aggregates nanoparticles organize in liquid condensed phase whereas small aggregates are localized in both fluid liquid-expanded (LE) and liquid-condenced (LC). We demonstrated also by atomic force microscopy that by measuring friction forces it is possible to get information as if nanoparticle aggregates are recovered or not by lipid monolayers and bilayers.
Abstract: Since hyaluronic acid (HA) receptor such as CD44 is
over-expressed at sites of cancer cells, HA can be used as a targeting
vehicles for anti-cancer drugs. The aim of this study is to synthesize
block copolymer composed of hyaluronic acid and
poly(ε-caprolactone) (HAPCL) and to fabricate polymeric micelles for
anticancer drug targeting against CD44 receptor of tumor cells.
Chemical composition of HAPCL was confirmed using 1H NMR
spectroscopy. Doxorubicin (DOX) was incorporated into polymeric
micelles of HAPCL. The diameters of HAPHS polymeric micelles
were changed around 80nm and have spherical shapes. Targeting
potential was investigated using CD44-overexpressing. When
DOX-incorporated polymeric micelles was added to KB cells, they
revealed strong red fluorescence color while blocking of CD44
receptor by pretreatment of free HA resulted in reduced intensity,
indicating that HAPCL polymeric micelles have targetability against
CD44 receptor.