Abstract: In vitro conservation of orchid germplasm provides an effective technique for ex situ conservation of orchid diversity. In this study, an efficient protocol for in vitro conservation of Vandopsis lissochiloides (Gaudich.) Pfitz. plantlet under slow growth conditions was investigated. Plantlets were cultured on different strength of Vacin and Went medium (½VW and ¼VW) supplemented with different concentrations of mannitol (0, 2, 4, 6 and 8%), sucrose (0 and 3%) and 50 g/L potato extract, 150 mL/L coconut water. The cultures were incubated at 25±2 °C and maintained under 20 µmol/m2s light intensity for 24 weeks without subculture. At the end of preservation period, the plantlets were subcultured to fresh medium for growth recovery. The results found that the highest leaf number per plantlet could be observed on ¼VW medium without adding sucrose and mannitol while the highest root number per plantlet was found on ½VW added with 3% sucrose without adding mannitol after 24 weeks of in vitro storage. The results showed that the maximum number of leaves (5.8 leaves) and roots (5.0 roots) of preserved plantlets were produced on ¼VW medium without adding sucrose and mannitol. Therefore, ¼VW medium without adding sucrose and mannitol was the best minimum growth conditions for medium-term storage of V. lissochiloides plantlets.
Abstract: Target of this study was the analysis of the impact of
crude glycerol on canine spermatozoa motility, morphology,
viability, and membrane integrity. Experiments were realized in vitro.
In the study, semen from 5 large dog breeds was used. They were
typical representatives of large breeds, coming from healthy rearing,
regularly vaccinated and integrated to the further breeding. Semen
collections were realized at the owners of animals and in the
veterinary clinic. Subsequently the experiments were realized at the
Department of Animal Physiology of the SUA in Nitra. The
spermatozoa motility was evaluated using CASA analyzer
(SpermVisionTM, Minitub, Germany) at the temperature 5 and 37°C
for 5 hours. In the study, 13 motility parameters were evaluated.
Generally, crude glycerol has generally negative effect on
spermatozoa motility. Morphological analysis was realized using
Hancock staining and the preparations were evaluated at
magnification 1000x using classification tables of morphologically
changed spermatozoa. Data clearly detected the highest number of
morphologically changed spermatozoa in the experimental groups
(know twisted tails, tail torso and tail coiling). For acrosome
alterations swelled acrosomes, removed acrosomes and acrosomes
with undulated membrane were detected. In this study also the effect
of crude glycerol on spermatozoa membrane integrity were analyzed.
The highest crude glycerol concentration significantly affects
spermatozoa integrity. Results of this study show that crude glycerol
has effect of spermatozoa motility, viability, and membrane integrity.
Detected changes are related to crude glycerol concentration,
temperature, as well as time of incubation.
Abstract: More than 3000 plants of notable phyto-therapeutic
value grow in South Africa; these include Cissampelos capensis,
commonly known in Afrikaans as dawidjie or dawidjiewortel. C.
capensis is the most significant and popular medicinal plant used by
the Khoisan as well as other rural groups in the Western region of
South Africa. Its rhizomes are traditionally used to treat male fertility
problems. Yet, no studies have investigated the effects of this plant or
its extracts on human spermatozoa. Therefore, this study aimed at
investigating the effects of C. capensis rhizome extract (CRE)
fractions on ejaculated human spermatozoa in vitro. Spermatozoa
from a total of 77 semen samples were washed with human tubular
fluid medium supplemented with bovine serum albumin (HTF-BSA)
and incubated for 2 hours with 20 μg/ml progesterone (P4) followed
by incubation with different concentrations (0, 0.05, 0.5, 5, 50, 200
μg/ml) of fractionated CRE (F1=0% MeOH, F2=30% MeOH,
F3=60% MeOH and F4=100% MeOH) for 1.5 hours at 37°C. A
sample without addition of CRE fractions served as control. Samples
were analyzed for sperm motility, reactive oxygen species (ROS),
DNA-fragmentation, acrosome reaction and capacitation. Results
showed that F1 resulted in significantly higher values for ROS,
capacitation and hyper-activation compared to F2, F3, and F4 with
P4-stimulated samples generally having higher values. No significant
effect was found for the other parameters. In conclusion, alkaloids
present in F1 of CRE appear to have triggered sperm intrinsic ROS
production leading to sperm capacitation and acrosome reaction
induced by P4.
Abstract: Amyloid aggregation of polypeptides is related to a
growing number of pathologic states known as amyloid disorders. In
recent years, blocking or reversing amyloid aggregation via the use of
small compounds are considered as two useful approaches in
hampering the development of these diseases. In this research, we
have compared the ability of several manganese-salen derivatives, as
synthetic compounds, and apigenin, as a natural flavonoid, to inhibit
of hen egg-white lysozyme (HEWL) aggregation, as an in vitro
model system.
Different spectroscopic analyses such as Thioflavin T (ThT) and
Anilinonaphthalene-8-sulfonic acid (ANS) fluorescence, Congo red
(CR) absorbance along with transmission electron microscopy were
used in this work to monitor the HEWL aggregation kinetic and
inhibition. Our results demonstrated that both type of compounds
were capable to prevent the formation of lysozyme amyloid
aggregation in vitro. In addition, our data indicated that synthetic
compounds had higher activity to inhibit of the β-sheet structures
relative to natural compound. Regarding the higher antioxidant
activities of the salen derivatives, it can be concluded that in addition
to aromatic rings of each of the compounds, the potent antioxidant
properties of salen derivatives contributes to lower lysozyme fibril
accumulation.
Abstract: Probiotic bacteria especially Lactobacillus spp. and Bifidobacterium exert suppressive effect on Helicobacter pylori. Cinnamon and licorice have been traditionally used for the treatment of gastric ulcer. The objectives of this study were to determine the effects of herbs on yogurt fermentation, the level of probiotic bacteria in yogurt during 28 days storage and the effect of herbal yogurt on the growth of H. pylori in vitro. Cinnamon or licorice was mixed with milk and the mixture was fermented with probiotic bacteria to form herbal-yogurt. Changes of pH and total titratable acids were monitored and the viability of probiotic bacteria was evaluated during and after refrigerated storage. The in vitro inhibition of H. pylori growth was determined using agar diffusion and minimum inhibitory concentration (MIC) method. The presence of herbs did not affect the probiotic population during storage. There were no significant differences in pH and TTA between herbal-yogurts and plain-yogurt during fermentation and storage. Water extract of cinnamon-yogurt showed the highest inhibition effect (13.5mm) on H. pylori growth in comparison with licorice-yogurt (11.2mm). The present findings indicate cinnamon and licorice has bioactive components to decrease the growth of H. pylori.
Abstract: In mammalian reproductive tract, the oviduct secretes
huge number of growth factors and cytokines that create an optimal
micro-environment for the initial stages of preimplantation embryos.
Secretion of these growth factors is stage-specific. Among them,
VEGF is a potent mitogen for vascular endothelium and stimulates
vascular permeability. Apart from angiogenesis, VEGF in the oviduct
may be involved in regulating the oocyte maturation and subsequent
developmental process during embryo production in vitro. In
experiment 1, to evaluate the effect of VEGF during IVM of porcine
COC and subsequent developmental ability after PA and SCNT. The
results from these experiments indicated that maturation rates among
the different VEGF concentrations were not significant different. In
experiment 2, total intracellular GSH concentrations of oocytes
matured with VEGF (5-50 ng/ml) were increased significantly
compared to a control and VEGF group (500 ng/ml). In experiment 3,
the blastocyst formation rates and total cell number per blastocyst
after parthenogenesis of oocytes matured with VEGF (5-50 ng/ml)
were increased significantly compared to a control and VEGF group
(500 ng/ml). Similarly, in experiment 4, the blastocyst formation rate
and total cell number per blastocyst after SCNT and IVF of oocytes
matured with VEGF (5 ng/ml) were significantly higher than that of
oocytes matured without VEGF group. In experiment 5, at 10 hour
after the onset of IVF, pronuclear formation rate was evaluated.
Monospermy was significantly higher in VEGF-matured oocytes than
in the control, and polyspermy and sperm penetration per oocyte
were significantly higher in the control group than in the VEGFmatured
oocytes. Supplementation with VEGF during IVM
significantly improved male pronuclear formation as compared with
the control. In experiment 6, type III cortical granule distribution in
oocytes was more common in VEGF-matured oocytes than in the
control. In conclusion, the present study suggested that
supplementation of VEGF during IVM may enhance the
developmental potential of porcine in vitro embryos through increase
of the intracellular GSH level, higher MPN formation and increased
fertilization rate as a consequence of an improved cytoplasmic
maturation.
Abstract: In this research, the main aim is to investigate the
antimicrobial effectiveness of ammonyx solutions finishing on
Sweatshirt Sport with immersion method. 60 Male healthy subjects
(football player) participated in this study. They were dressed in a
Sweatshirt for 14 days and some microbes found on them were
investigated. The antimicrobial effect of different ammonyx
solutions(1/100, 1/500, 1/1000, 1/2000 v/v solutions of Ammonyx)
on the identified microbes was studied by the zone inhabitation
method in vitro. In the next step the Sweatshirt Sports were treated
with the same different solutions of ammonyx and the antimicrobial
effectiveness was assessed by colony count method in different times
and the results were compared whit untreated ones. Some mechanical
properties of treated cotton/polyester yarn that used in Sweatshirt
Sport were measured after 30 days and were compared with
untreated one. Finally after finishing, scanning electron microscopy
(SEM) was used to compare the surfaces of the finished and
unfinished specimens. The results showed the presence of five
pathogenic microbes on Sweatshirt Sports such as Escherichia coli,
Staphylococcus aureus, Aspergillus, Mucor and Candida. The
inhalation time for treated on Sweatshirt Sports improved. The
amount of colony growth on treated clothes reduced considerably
and moreover the mechanical tests results showed no significant
deterioration effect of studies properties in comparison to the
untreated yarn. The visual examination of the SEM indicated that the
antimicrobial treatments were applied usefully to fabrics.
Abstract: Transpedicular screw fixation in spinal fractures,
degenerative changes, or deformities is a well-established procedure.
However, important rate of fixation failure due to screw bending,
loosening, or pullout are still reported particularly in weak bone stock
in osteoporosis. To overcome the problem, mechanism of failure has
to be fully investigated in vitro. Post-mortem human subjects are less
accessible and animal cadavers comprise limitations due to different
geometry and mechanical properties. Therefore, the development of a
synthetic model mimicking the realistic human vertebra is highly
demanded. A bone surrogate, composed of Polyurethane (PU) foam
analogous to cancellous bone porous structure, was tested for 3
different densities in this study. The mechanical properties were
investigated under uniaxial compression test by minimizing the end
artifacts on specimens. The results indicated that PU foam of 0.32
g.cm-3 density has comparable mechanical properties to human
cancellous bone in terms of young-s modulus and yield strength.
Therefore, the obtained information can be considered as primary
step for developing a realistic cancellous bone of human vertebral
body. Further evaluations are also recommended for other density
groups.
Abstract: Magnesium alloy has been widely investigated as
biodegradable cardiovascular stent and bone implant. Its application
for biodegradable esophageal stenting remains unexplored. This
paper reports the biodegradation behaviors of AZ31 magnesium alloy
in artificial saliva and various types of beverage in vitro. Results
show that the magnesium ion release rate of AZ31 in artificial saliva
for a stent (2cm diameter, 10cm length at 50% stent surface
coverage) is 43 times lower than the daily allowance of human body
magnesium intakes. The degradation rates of AZ31 in different
beverages could also be significantly different. These results suggest
that the esophagus in nature is a less aggressive chemical
environment for degradation of magnesium alloys. The significant
difference in degradation rates of AZ31 in different beverages opens
new opportunities for development of degradation controllable
esophageal stent through customizing ingested beverages.
Abstract: It has been shown that pH 7,3 and 37 0C are the optimal condition for the growth of E. coli “ASAP". The cells grow well on Glucose, Lactose, D-Mannitol, D-Sorbitol, (+)-Xylose, L- (+)-Arabinose and Dulcitol. No growth has been observed on Sucrose, Inositol, Phenylalanine, and Tryptophan. The strain is sensitive to a range of antibiotics. The present study has demonstrated that E. coli “ASAP" inhibit the growth of S. enterica ATCC #700931 in vitro. The studies on conjugating activity has revealed no conjugant of E. coli “ASAP" with plasmid strains E. coli G35#59 and S. enterica ATCC #700931. On the other hand, the conjugants with low frequencies were obtained from E. coli “ASAP" with E. coli G35#61, and E. coli “ASAP" with randomly chosen isolate from healthy human gut microflora: E. coli E6. The results of present study have demonstrated improvements in gut microflora condition of patients with different diseases after the administration of “ASAP"
Abstract: The importance for manipulating an incorporated
scaffold and directing cell behaviors is well appreciated for tissue
engineering. Here, we developed newly nano-topographic oxidized
silicon nanosponges capable of being various chemical modifications
to provide much insight into the fundamental biology of how cells
interact with their surrounding environment in vitro. A wet etching
technique is exerted to allow us fabricated the silicon nanosponges in a
high-throughput manner. Furthermore, various organo-silane
chemicals enabled self-assembled on the surfaces by vapor deposition.
We have found that Chinese hamster ovary (CHO) cells displayed
certain distinguishable morphogenesis, adherent responses, and
biochemical properties while cultured on these chemical modified
nano-topographic structures in compared with the planar oxidized
silicon counterparts, indicating that cell behaviors can be influenced
by certain physical characteristic derived from nano-topography in
addition to the hydrophobicity of contact surfaces crucial for cell
adhesion and spreading. Of particular, there were predominant
nano-actin punches and slender protrusions formed while cells were
cultured on the nano-topographic structures. This study shed potential
applications of these nano-topographic biomaterials for controlling
cell development in tissue engineering or basic cell biology research.
Abstract: Deaminated lesions were produced via nitrosative oxidation of natural nucleobases; uracul (Ura, U) from cytosine (Cyt, C), hypoxanthine (Hyp, H) from adenine (Ade, A), and xanthine (Xan, X) and oxanine (Oxa, O) from guanine (Gua, G). Such damaged nucleobases may induce mutagenic problems, so that much attentions and efforts have been poured on the revealing of their mechanisms in vivo or in vitro. In this study, we employed these deaminated lesions as useful probes for analysis of DNA-binding/recognizing proteins or enzymes. Since the pyrimidine lesions such as Hyp, Oxa and Xan are employed as analogues of guanine, their comparative uses are informative for analyzing the role of Gua in DNA sequence in DNA-protein interaction. Several DNA oligomers containing such Hyp, Oxa or Xan substituted for Gua were designed to reveal the molecular interaction between DNA and protein. From this approach, we have got useful information to understand the molecular mechanisms of the DNA-recognizing enzymes, which have not ever been observed using conventional DNA oligomer composed of just natural nucleobases.
Abstract: To identify an endothelial cell-specific promoter suitable for vascular-specific targeting, we tested five promoters in vitro--Tie2SE, Tie2LE, ICAM2, Flt-1 and vWF--for promoter activity and specificity in endothelial cells, smooth muscle cells and non-vascular resident cells as well as tissues. These promoters, except for vWF, exhibited good endothelial activity and specificity in vitro. In a syngenic heart transplantation model, the ICAM2 promoter was variably functional in coronary endothelial cells of donor hearts. Thus, the ICAM2, Flt-1, Tie2SE and Tie2LE promoters hold promise for endothelial-specific targeting, but in vitro expression may not predict in vivo expression.
Abstract: Leonotisleonurus a shrub indigenous to Southern
Africa is widely used in traditional medicine to treat a variety of
conditions ranging from skin diseases and cough to epileptic fits and
‘heart problems’. Studies on the aqueous extract of the leaves have
indicated cycloxegenase enzyme inhibitory activity and an
antihypertensive effect.
Five methanol leaf extract fractions (MLEa - MLEe) of L.
leonurus were tested on anaesthetized normotensive male Wistar rats
(AWR) and isolated perfused working rat hearts (IWH). Fraction
MLEc (0.01mg/kg – 0.05mg/kg) induced significant increases in BP
and HR in AWR and positive chronotropic and inotropic effects in
IWH (1.0mg/ml – 5.0mg/ml). Pre-administration of atenolol
(2.0mg/kg) and prazosin (60μg/kg) significantly inhibited MLEc
effect on HR and MAP respectively in vivo, while atenolol
(7.0mg/ml) pre-perfusion significantly inhibited MLEc effect in vitro.
The hypertensive effect of MLEc is probably via β1agonism.
Results also indicate the presence of multiple cardioactive
compounds in L. leonurus.