Abstract: In vitro plant regeneration has been successfully obtained from basal shoot explant of Vetiveria zizanioides through indirect organogenesis. The explant was cultured in Murashige & Skoog’s (MS) media supplemented with 2,4-D, IAA, and kinetin in various concentrations. Callus was well induced in media supplemented with 2 ppm 2,4-D, 1 ppm IAA, and 1 ppm kinetin. This callus was then transferred to MS media supplemented with 1 - 5 ppm of BAP for shoot regeneration. The media supplemented with 3 ppm BAP was a suitable medium for shoot induction, as well as for shoot multiplication. Rooting was well developed in shoot following transferred to half MS media containing 0.2 ppm IBA. Plantlet was then transferred to husk charcoal for acclimatization, and almost all (90%) of plantlets were survived during acclimatization.
Abstract: The methanolic extracts from seeds of tamarind
(Tamarindus indica) was prepared by Soxhlet apparatus extraction
and evaluated for total phenolic content by Folin-Ciocalteu method.
Then, methanolic extract was screened biological activities (In vitro)
for anti-melanogenic activity by tyrosinase inhibition test, antiinflammation
activity by cyclooxygenase 1 (COX-1) and
cyclooxygenase 2 (COX-2) inhibition test, and cytotoxic screening
test with Vero cells. The results showed that total phenolic content,
which contained in extract, was contained 27.72 mg of gallic acid
equivalent per g of dry weight. The ability to inhibit tyrosinase
enzyme, which exerted by Tamarind seed extracts (1 mg/ml) was
52.13 ± 0.42 %. The extract was not possessed inhibitory effect to
COX-1 and COX-2 enzymes and cytotoxic effect to Vero cells. The
finding is concludes that tested seed extract was possessed
antimelanogenic activity with non-toxic effects. However, there was
not exhibited anti-inflammatory activity. Further studies include the
use of advance biological models to confirm this biological activity,
as well as, the isolation and characterization of the purified
compounds that it was contained.
Abstract: Heat-inducible gene expression vectors are useful for hyperthermia-induced cancer gene therapy, because the combination
of hyperthermia and gene therapy can considerably improve the therapeutic effects. In the present study, we developed an enhanced
heat-inducible transgene expression system in which a heat-shock
protein (HSP) promoter and tetracycline-responsive transactivator
were combined. When the transactivator plasmid containing the
tetracycline-responsive transactivator gene was co-transfected with
the reporter gene expression plasmid, a high level of heat-induced gene expression was observed compared with that using the HSP
promoter without the transactivator. In vitro evaluation of the
therapeutic effect using HeLa cells showed that heat-induced therapeutic gene expression caused cell death in a high percentage of
these cells, indicating that this strategy is promising for cancer gene therapy.
Abstract: Lectins have a good scope in current clinical
microbiology research. In the present study evaluated the
antimicrobial activities of a D-galactose binding lectin (PnL) was
purified from the annelid, Perinereis nuntia (polychaeta) by affinity
chromatography. The molecular mass of the lectin was determined to
be 32 kDa as a single polypeptide by SDS-PAGE under both reducing
and non-reducing conditions. The hemagglutinating activity of the
PnL showed against trypsinized and glutaraldehyde-fixed human
erythrocytes was specifically inhibited by D-Gal, GalNAc,
Galβ1-4Glc and Galα1-6Glc. PnL was evaluated for in vitro
antibacterial screening studies against 11 gram-positive and
gram-negative microorganisms. From the screening results, it was
revealed that PnL exhibited significant antibacterial activity against
gram-positive bacteria. Bacillus megaterium showed the highest
growth inhibition by the lectin (250 μg/disc). However, PnL did not
inhibit the growth of gram-negative bacteria such as Vibrio cholerae
and Pseudomonas sp. PnL was also examined for in vitro antifungal
activity against six fungal phytopathogens. PnL (100 μg/mL) inhibited
the mycelial growth of Alternaria alternata (24.4%). These results
indicate that future findings of lectin applications obtained from
annelids may be of importance to life sciences.
Abstract: F-actin fibrils are the cytoskeleton of osteocytes. They react in a dynamic manner to mechanical loading, and strength and
reposition their efforts to reinforce the cells structure. We hypothesize that f-actin is temporarly disrupted after loading and repolymerizes
in a new orientation to oppose the applied load. In vitro studies are conducted to determine f-actin disruption after varying mechanical stimulus parameters that are known to affect bone
formation. Results indicate that the f-actin cytoskeleton is disrupted in vitro as a function of applied mechanical stimulus parameters and
that the f-actin bundles reassemble after loading induced disruption
within 3 minutes after cessation of loading. The disruption of the factin
cytoskeleton depends on the magnitude of stretch, the numbers
of loading cycles, frequency, the insertion of rest between loading
cycles and extracellular calcium. In vivo studies also demonstrate
disruption of the f-actin cytoskeleton in cells embedded in the bone
matrix immediately after mechanical loading. These studies suggest
that adaptation of the f-actin fiber bundles of the cytoskeleton in
response to applied loads occurs by disruption and subsequent repolymerization.
Abstract: The present study has been taken to explore the
screening of in vitro antimicrobial activities of D-galactose-binding
sponge lectin (HOL-30). HOL-30 was purified from the marine
demosponge Halichondria okadai by affinity chromatography. The
molecular mass of the lectin was determined to be 30 kDa with a
single polypeptide by SDS-PAGE under non-reducing and reducing
conditions. HOL-30 agglutinated trypsinized and glutaraldehydefixed
rabbit and human erythrocytes with preference for type O
erythrocytes. The lectin was subjected to evaluation for inhibition of
microbial growth by the disc diffusion method against eleven human
pathogenic gram-positive and gram-negative bacteria. The lectin
exhibited strong antibacterial activity against gram-positive bacteria,
such as Bacillus megaterium and Bacillus subtilis. However, it did
not affect against gram-negative bacteria such as Salmonella typhi
and Escherichia coli. The largest zone of inhibition was recorded of
Bacillus megaterium (12 in diameter) and Bacillus subtilis (10 mm in
diameter) at a concentration of the lectin (250 μg/disc). On the other
hand, the antifungal activity of the lectin was investigated against six
phytopathogenic fungi based on food poisoning technique. The lectin
has shown maximum inhibition (22.83%) of mycelial growth of
Botrydiplodia theobromae at a concentration of 100 μg/mL media.
These findings indicate that the lectin may be of importance to
clinical microbiology and have therapeutic applications.
Abstract: Nigella sativa L. is an aromatic plant belonging to the
family Ranunculaceae. It has been used traditionally, especially in the
middle East and India, for the treatment of asthma, cough, bronchitis,
headache, rheumatism, fever, influenza and eczema. Several
biological activities have been reported in Nigella sativa seeds,
including antioxidant. In this context we tried to estimate the
antioxidant activity of various extracts prepared from Nigella sativa
seeds, methanolic extract (ME), chloroformic extract (CE), hexanic
extract (HE : fixed oil), ethyl acetate extract (EAE) water extract
(WE). The Folin-Ciocalteu assay showed that CE and EAE contained
high level of phenolic compounds 81.31 and 72.43μg GAE/mg of
extract respectively. Similarly, the CE and EAE exhibited the highest
DPPH radical scavenging activity, with IC50 values of 106.56μg/ml
and 121.62μg/ml respectively. In addition, CE and HE showed the
most scavenging activity against superoxide radical generated in the
PMS-NADH-NBT system with respective IC50 values of 361.86
μg/ml and 371.80 μg/ml, which is comparable to the activity of the
standard antioxidant BHT (344.59 μg/ml). Ferrous ion chelating
capacity assay showed that WE, EAE and ME are the most active
with 40.57, 39.70 and 22.02 mg EDTA-E/g of extract. The inhibition
of linoleic acid/ß-carotene coupled oxidation was estimated by ßcarotene
bleaching assay, this showed a highest relative antioxidant
activity with CE and EAE (69.82% of inhibition). The antioxidant
activities of the methanolic extract and the fixed oil are confirmed by
an in vivo assay in mice, the daily oral administration of methanolic
extract (500 and 800 mg/kg/day) and fixed oil (2 and 4 ml/kg/day)
during 21 days, resulted in a significant enhancement of the blood
total antioxidant capacity (measured by KRL test) and the plasmatic
antioxidant capacity towards DPPH radical.
Abstract: Studies were carried out to determine the in vitro
susceptibility of the typhoid pathogens to combined action of Euphorbia hirta, Euphorbia heterophylla and Phyllanthus niruri. Clinical isolates of the typhoid bacilli were subjected to susceptibility testing using agar diffusion technique and the minimum inhibitory
concentration (MIC) determined with tube dilution technique. These
isolates, when challenged with doses of the extracts from the three
medicinal plants showed zones of inhibition as wide as 26±0.2mm, 22±0.1mm and 18±0.0mm respectively. The minimum inhibitory concentration (MIC) revealed organisms inhibited at varying
concentrations of extracts: E. hirta (S. typhi 0.250mg/ml, S. paratyphi A 0.125mg/ml, S. paratyphi B 0.185mg/ml and S. paratyphi C 0.225mg/ml), E. heterophylla (S. typhi 0.280mg/ml, S. paratyphi A
0.150mg/ml, S. paratyphi B 0.200mg/ml and S. paratyphi C 0.250mg/ml) and P. niruri (S. typhi 0.150mg/ml, S. paratyphi A 0.100mg/ml, S. paratyphi B 0.115mg/ml and S. paratyphi C 0.125mg/ml). The results of the synergy between the three plants in
the ration of 1:1:1 showed very low MICs for the test pathogens as follows S. typhi 0.025mg/ml, S. paratyphi A 0.080mg/ml, S. paratyphi B 0.015mg/ml and S. paratyphi C 0.10mg/ml with the
diameter zone of inhibition (DZI) ranging from 35±0.2mm,
28±0.4mm, 20±0.1mm and 32±0.3mm respectively. The secondary
metabolites were identified using simple methods and HPLC. Organic components such as anthroquinones, different alkaloids,
tannins, 6-ethoxy-1,2,3,4-tetrahydro-2,2,4-trimethyl and steroids were identified. The prevalence of Salmonellae, a deadly infectious disease, is still very high in parts of Nigeria. The synergistic action of these three plants is very high. It is concluded that pharmaceutical companies should take advantage of these findings to develop new
anti-typhoid drugs from these plants.
Abstract: DNA shuffling is a powerful method used for in vitro
evolute molecules with specific functions and has application in areas
such as, for example, pharmaceutical, medical and agricultural
research. The success of such experiments is dependent on a variety
of parameters and conditions that, sometimes, can not be properly
pre-established. Here, two computational models predicting DNA
shuffling results is presented and their use and results are evaluated
against an empirical experiment. The in silico and in vitro results
show agreement indicating the importance of these two models and
motivating the study and development of new models.
Abstract: The paper presents the optimization problem for the
multi-element synthetic transmit aperture method (MSTA) in
ultrasound imaging applications. The optimal choice of the transmit
aperture size is performed as a trade-off between the lateral
resolution, penetration depth and the frame rate. Results of the
analysis obtained by a developed optimization algorithm are
presented. Maximum penetration depth and the best lateral resolution
at given depths are chosen as the optimization criteria. The
optimization algorithm was tested using synthetic aperture data of
point reflectors simulated by Filed II program for Matlab® for the
case of 5MHz 128-element linear transducer array with 0.48 mm
pitch are presented. The visualization of experimentally obtained
synthetic aperture data of a tissue mimicking phantom and in vitro
measurements of the beef liver are also shown. The data were
obtained using the SonixTOUCH Research systemequipped with a
linear 4MHz 128 element transducerwith 0.3 mm element pitch, 0.28
mm element width and 70% fractional bandwidth was excited by one
sine cycle pulse burst of transducer's center frequency.
Abstract: Liposomal magnetofection is the most powerful nonviral method for the nucleic acid delivery into the cultured cancer cells and widely used for in vitro applications. Use of the static magnetic field condition may result in non-uniform distribution of aggregate complexes on the surface of cultured cells. To prevent this, we developed the new device which allows to concentrate aggregate complexes under dynamic magnetic field, assisting more contact of these complexes with cellular membrane and, possibly, stimulating endocytosis. Newly developed device for magnetofection under dynamic gradient magnetic field, “DynaFECTOR", was used to compare transfection efficiency of human liver hepatocellular carcinoma cell line HepG2 with that obtained by lipofection and magnetofection. The effect of two parameters on transfection efficiency, incubation time under dynamic magnetic field and rotation frequency of magnet, was estimated. Liposomal magnetofection under dynamic gradient magnetic field showed the highest transfection efficiency for HepG2 cells.
Abstract: Today, cancer remains one of the major diseases that
lead to death. The main obstacle in chemotherapy as a main cancer
treatment is the toxicity to normal cells due to Multidrug Resistance
(MDR) after the use of anticancer drugs. Proposed solution to
overcome this problem is the use of MDR efflux inhibitor of cinchona
alkaloids which is delivered together with anticancer drugs
encapsulated in the form of polymeric nanoparticles. The particles
were prepared by the hydration method. The characterization of
nanoparticles was particle size, zeta potential, entrapment efficiency
and in vitro drug release. Combination nanoparticle size ranged 29-45
nm with a neutral surface charge. Entrapment efficiency was above
87% for the use quinine, quinidine or cinchonidine in combination
with etoposide. The release test results exhibited that the cinchona
alkaloids release released faster than that of etoposide. Collectively,
cinchona alkaloids can be packaged along with etoposide in
nanomicelles for better cancer therapy.
Abstract: Bone remodeling occurs by the balanced action of
bone resorbing osteoclasts (OC) and bone-building osteoblasts.
Increased bone resorption by excessive OC activity contributes
to malignant and non-malignant diseases including osteoporosis.
To study OC differentiation and function, OC formed in
in vitro cultures are currently counted manually, a tedious
procedure which is prone to inter-observer differences. Aiming
for an automated OC-quantification system, classification of
OC and precursor cells was done on fluorescence microscope
images based on the distinct appearance of fluorescent nuclei.
Following ellipse fitting to nuclei, a combination of eight
features enabled clustering of OC and precursor cell nuclei.
After evaluating different machine-learning techniques, LOGREG
achieved 74% correctly classified OC and precursor cell
nuclei, outperforming human experts (best expert: 55%). In
combination with the automated detection of total cell areas,
this system allows to measure various cell parameters and most
importantly to quantify proteins involved in osteoclastogenesis.
Abstract: Biologically active peptides are of particular interest
in food science and human nutrition because they have been shown to play several physiological roles. In vitro gastrointestinal digestion of lentil and whey proteins in this study produced high angiotensin-I converting enzyme inhibitory activity with 75.5±1.9 and 91.4±2.3%
inhibition, respectively. High ACE inhibitory activity was observed in lentil after 5 days of germination (84.3±1.2%). Fractionation by
reverse phase chromatography gave inhibitory activities as high as
86.3±2.0 for lentil, 94.8±1.8% for whey and 93.7±1.7% at 5th day of germination. Further purification by HPLC resulted in several
inhibitory peptides with IC50 values ranging from 0.064 to 0.164
mg/ml. These results demonstrate that lentil proteins are a good
source of peptides with ACE inhibitory activity that can be released by germination or gastrointestinal digestion. Despite the lower bioactivity in comparison with whey proteins, incorporation of lentil proteins in functional food formulations and natural drugs look promising.