Adhesion Properties of Bifidobacterium Pseudocatenulatum G4 and Bifidobacterium Longum BB536 on HT-29 Human Epithelium Cell Line at Different Times and pH

Adhesion to the human intestinal cell is considered as one of the main selection criteria of lactic acid bacteria for probiotic use. The adhesion ability of two Bifidobacteriums strains Bifidobacterium longum BB536 and Bifidobacterium psudocatenulatum G4 was done using HT-29 human epithelium cell line as in vitro study. Four different level of pH were used 5.6, 5.7, 6.6, and 6.8 with four different times 15, 30, 60, and 120 min. Adhesion was quantified by counting the adhering bacteria after Gram staining. The adhesion of B. longum BB536 was higher than B. psudocatenulatum G4. Both species showed significant different in the adhesion properties at the factors tested. The highest adhesion for both Bifidobacterium was observed at 120 min and the low adhesion was in 15 min. The findings of this study will contribute to the introduction of new effective probiotic strain for future utilization.

Effects of a Nectandra Membranacea Extract on Labeling of Blood Constituents with Technetium-99m and on the Morphology of Red Blood Cells

The aim of this in vitro study was to evaluate the possible interference of a Nectandra membranacea extract (i) on the labeling of blood cells (BC), (ii) on the labeling process of BC and plasma (P) proteins with technetium-99m (Tc-99m) and (iii) on the morphology of red blood cells (RBC). Blood samples were incubated with a Nectandra membranacea crude extract, stannous chloride, Tc- 99m (sodium pertechnetate) was added, and soluble (SF) and insoluble (IF) fractions were isolated. Morphometry studies were performed with blood samples incubated with Nectandra membranacea extract. The results show that the Nectandra membranacea extract does not promote significant alteration of the labeling of BC, IF-P and IF-BC. The Nectandra membranacea extract was able to alter the erythrocyte membrane morphology, but these morphological changes were not capable to interfere on the labeling of blood constituents with Tc-99m.

Adding Olive Oil into Diluents for Improving Semen Quality and Storage Ability of Roosters' Semen during Liquid Storage

The aim of this study was to investigate the effects of supplementing the diluent of roosters' semen with different levels of olive oil on motility, viability, morphology and acrosome integrity of chicken spermatozoa after in vitro storage for up to 72 h. Semen was collected from 60 White Layer males (62 wk of age) kept in separated floor pens and randomly divided into six treatment groups (10 males in each group). Experimental groups were as follows: T1 :fresh semen, T2 : semen extended 1:1 with Al – Daraji 2 diluent (AD2D) alone, T3 – T6 :semen samples extended 1:1 with AD2D supplemented with 2 ml, 4 ml, 6 ml or 8 ml of olive oil / 100 ml of diluent, respectively. Semen samples were then stored at 5 °C for 24 h, 48 h or 72 h. There was a clear influence of diluent supplementation with olive oil on the spermatozoa motility profile; olive oil groups (T3, T4, T5 and T6) recorded the highest scores of mass activity and individual motility during all storage periods compared to T1 and T2 groups. In addition, the inclusion of olive oil into semen diluent (T3, T4, T5 and T6) gave significantly higher percentages of viable spermatozoa, normal morphologically spermatozoa and intact acrosomes irrespective of storage period. These results clearly show that supplementation the diluent of roosters' semen with olive oil can improve semen quality when semen samples in vitro stored at 5 °C for up to 72 h.

Effect of Genotype, Explant Type and Growth Regulators on The Accumulation of Flavonoides of (Silybum marianum L.) in In vitro Culture

The extract of milk thistle contains a mix of flavonolignans termed silymarine.. In order to analysis influence of growth regulators, genotype, explant and subculture on the accumulation of flavonolignans, a study was carried out by using two genotype (Budakalszi and Noor abad moghan cultivars), cotyledon and hypocotyle explants, solid media of MS supplemented by different combinations of two growth regulators; Kinetin (0.1, 1 mg/l) and 2,4-D (1, 2 mg/l). Seeds of the plant were germinated in MS media whitout growth regulators in growth chamber at 26°C and darkness condition. In order to callus induction, the culture media was supplemented whit different concentrations of 2,4-D and kinetin. Calli obtained from explants were sub-cultured four times into the fresh media of the first experiment. flavonoides was extracted from calli in four subcultures. The flavonoid components were determined by high- performance liquid choromatography (HPLC) and separated into Taxifolin, Silydianin+Silychristin, Silybin A+B and Isosilybin A+B. Results showed that with increasing callus age, increased accumulation of silybin A+B, but reduced Isosilybin A+B content. Highest accumulation of Taxifolin was observed at first calli. Calli produced from cotyledon explant of Budakalszi cultivar were superior for Silybin A+B, where calli from hypocotyl explant produced higher amount of Taxifolin and Silydianin+Silychristin. The best cultivar for Silymarin production in this study was Budakalszi cultivar. High amount of SBN A+B and TXF were obtained from hypocotil explant.

Application of an in vitro Alveolus Model in Evaluating the Alveolar Response to Pressure- Induced Injury

In an effort to understand the preliminary effects of aerodynamic stress on alveolar epithelial cells, we developed a multifluidic cell culture platform capable of supporting alveolar cultures at an air-liquid interface under constant air flow and exposure to varying pressure stimuli on the apical side while providing nourishment on the basolateral plane. Our current study involved utilizing the platform to study the effect of basement membrane coating and addition of dexamethasone on cellular response to pressure in A549 and H441 alveolar epithelial cells.

Effect of White Kwao Extract (Pueraria mirifica) on in vitro Development and Implantation Rate of Mouse Embryo

The White Kwao (Pueraria mirifica), a potent phytoestrogenic medicinal plant, has long been use in Thailand as a traditional folkmedicine. However, no scientific information of the direct effect of White Kwao on the development of mammalian embryo was available. Therefore, the purpose of this study was to investigate the effect of White Kwao extract on the in vitro development and implantation rate of mouse embryos. This study was designed into two experiments. In the first experiment, the two-cell stage mouse embryos were collected from the oviduct of superovulated mature female mice, and randomly cultured in three different media, the M16, M16 supplemented with 0.52μg esthinylestradiol-17β, and M16 supplemented with 10 mg/ml White Kwao extract. The culture was incubated in CO2 incubator at 37 oC . After the embryos were cultivated, the developments of embryos were observed every 24 hours for 5 days. The development rate of embryos from the two-cell stage to blastocyst stage in the media was with White Kwao was significantly higher (p

Effects of Chitosan as the Growth Stimulator for Grammatophyllum speciosum in Vitro Culture

The effects of chitosan, a biodegradable polymer, were studied in Grammatophyllum speciosum protocorm-like bodies (PLBs) in vitro culture. The chitosan concentration of 0, 5, 10, 15, 20, 25, 50 or 100 mg/l were supplemented in half-strength Murashige and Skoog (1/2 MS) liquid or on agar media containing 2% (w/v) sucrose. The results showed that liquid medium supplemented with 15 mg/l chitosan showed the highest relative growth rate (7-fold increase) of PLBs. On 1/2 MS agar medium supplemented with 25 mg/l chitosan gave the highest relative growth rate (4-fold increase). The relative growth rate of G. speciosum PLBs on agar medium was significantly lower than that in liquid medium. Moreover, chitosan, supplemented to agar medium promoted shoot formation but not rooting. However, supplementation at too high a level, such as 100 mg/l can inhibit growth and kill PLBs.

Biochemical Characteristics of Sorghum Flour Fermented and/or Supplemented with Chickpea Flour

Sorghum flour was supplemented with 15 and 30% chickpea flour. Sorghum flour and the supplement were fermented at 35 oC for 0, 8, 16, and 24 h. Changes in pH, titrable acidity, total soluble solids, protein content, in vitro protein digestibility and amino acid composition were investigated during fermentation and/or after supplementation of sorghum flour with chickpea. The pH of the fermenting material decreased sharply with a concomitant increase in the titrable acidity. The total soluble solids remained unchanged with progressive fermentation time. The protein content of sorghum cultivar was found to be 9.27 and that of chickpea was 22.47%. The protein content of sorghum cultivar after supplementation with15 and 30% chickpea was significantly (P ≤ 0.05) increased to 11.78 and 14.55%, respectively. The protein digestibility also increased after fermentation from 13.35 to 30.59 and 40.56% for the supplements, respectively. Further increment in protein content and digestibility was observed when supplemented and unsupplemented samples were fermented for different periods of time. Cooking of fermented samples was found to increase the protein content slightly and decreased digestibility for both supplements. Amino acid content of fermented and fermented and cooked supplements was determined. Supplementation was found to increase the lysine and therionine content. Cooking following fermentation decreased lysine, isoleucine, valine and sulfur containg amino acids.

Development and in vitro Characterization of Self-nanoemulsifying Drug Delivery Systems of Valsartan

The present study is aim to prepare and evaluate the selfnanoemulsifying drug delivery (SNEDDS) system of a poorly water soluble drug valsartan in order to achieve a better dissolution rate which would further help in enhancing oral bioavailability. The present research work describes a SNEDDS of valsartan using labrafil M 1944 CS, Tween 80 and Transcutol HP. The pseudoternary phase diagrams with presence and absence of drug were plotted to check for the emulsification range and also to evaluate the effect of valsartan on the emulsification behavior of the phases. The mixtures consisting of oil (labrafil M 1944 CS) with surfactant (tween 80), co-surfactant (Transcutol HP) were found to be optimum formulations. Prepared formulations were evaluated for its particle size distribution, nanoemulsifying properties, robustness to dilution, self emulsication time, turbidity measurement, drug content and invitro dissolution. The optimized formulations are further evaluated for heating cooling cycle, centrifugation studies, freeze thaw cycling, particle size distribution and zeta potential were carried out to confirm the stability of the formed SNEDDS formulations. The prepared formulation revealed t a significant improvement in terms of the drug solubility as compared with marketed tablet and pure drug.

Why We Are Taller in the Morning than Going to Bed at Night – An in vivo and in vitro Study

Intradiscal and intervertebral pressure transducers were developed. They were used to map the pressures in the nucleus and within the annulus of the human spinal segments. Their stressrelaxation were recorded over a period of time for nucleus pressure, applied load, and peripherial strain against time. The results show that for normal discs, pressures in the nucleus are viscoelastic in nature with the applied compressive load. Mechanical strains which develop around the periphery of the vertebral body are also viscoelastic with the applied compressive load. Applied compressive load against time also shows viscoelastic behavior. However, annulus does not respond viscoelastically with the applied load. It showed a linear response to compressive loading.

In vitro Anti-tubercular Screening of Newly Synthesized Benzimidazole Derivatives

A series of 1-(1H-benzimidazol-2-yl)-3-(substituted phenyl)-2-propen-1-one were allowed to react with hydrazine hydrate and phenyl hydrazine in submitted reactions to get pyrazoline and phenyl pyrazoline derivatives. All the compounds entered for screening at the Tuberculosis Antimicrobial Acquisition and Coordinating Facility (TAACF) for their in vitro antibacterial activity against Mycobacterium tuberculosis H37Rv strain (ATCC 27294) using Microplate Alamar Blue Assay (MABA) susceptibility test. The results expressed as MIC (minimum inhibitory concentration) in μg/mL. Among the fifteen compounds, eight compounds were found to have MIC values less than 10 μg/mL. These were subjected for cytotoxicity assay in VERO cells to determine CC50 (cytotoxic concentration 50%) values and finally SI (Selectivity Index) were calculated. Compound (XV) 2-[5-(4- fluorophenyl)-1-phenyl-4,5-dihydro-1H-3-pyrazolyl]-1Hbenzimidazole was considered the best candidate of the series that could be a good starting point to develop new lead compounds in the fight against tuberculosis.

Changes of in vitro Cytokine Production induced by δ-Lactams

The aim of this work was to study the in vitro effects of δ-lactam 1 and its 4-chlorophenyl derivative 2, on the proliferative responses of human lymphocytes and Th1 and Th2 cytokine secretion. The possible protective role of vitamin E on intracellular stress oxidative induced by these compounds was also investigated. Peripheral blood lymphocytes were isolated using differential centrifugation on a density gradient of Histopaque. They were cultured with mitogen concanavalin A, vitamin E (10 μM) and with different concentrations of the compounds 1 and 2 (0.1 to 10 μM). Proliferation (MTT assay), IL-2, INFγ and IL-4 (Elisa kits), intracellular superoxide anion were determined. 1 and 2 were immunostimulant and increased cytokine secretion with a shift away from Th1 response to Th2. These properties were however accompanied by an increase in intracellular oxidative stress. The presence of vitamin E exhibited protective effects by reducing δ- lactam-induced superoxide anion generation in lymphocytes.

Multiple Shoot Formation of Paphiopedilum 'Delrosi'

Shoots, with three leaves, of Paphiopedilum 'Delrosi' were used as explants for multiple shoot induction. Modified Hyponex medium was supplemented with thidiazuron (TDZ), N6- benzyladenine (BA) or kinetin (Kn) alone and in combinations with 2,4-dichlorophenoxyacetic acid (2,4-D). All explants were cultured for 15 weeks. It was found that TDZ alone at the concentration of 0.45μM or in combination with 4.52μM 2,4-D and 8.88μM BA in combination with 13.56μM 2,4-D promoted multiple shoots. The highest shoot sprouting efficiencies (80.0, 90.0 and 80.0%) and new shoot numbers (1.5, 1.3 and 1.1) were obtained, respectively. Fresh weight, height, numbers of leaf and root of new shoots and initial explants were discussed.

Analysis of DNA-Recognizing Enzyme Interaction using Deaminated Lesions

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.

In Vitro Study of Coded Transmission in Synthetic Aperture Ultrasound Imaging Systems

In the paper the study of synthetic transmit aperture method applying the Golay coded transmission for medical ultrasound imaging is presented. Longer coded excitation allows to increase the total energy of the transmitted signal without increasing the peak pressure. Moreover signal-to-noise ratio and penetration depth are improved while maintaining high ultrasound image resolution. In the work the 128-element linear transducer array with 0.3 mm inter-element spacing excited by one cycle and the 8 and 16- bit Golay coded sequences at nominal frequency 4 MHz was used. To generate a spherical wave covering the full image region a single element transmission aperture was used and all the elements received the echo signals. The comparison of 2D ultrasound images of the tissue mimicking phantom and in vitro measurements of the beef liver is presented to illustrate the benefits of the coded transmission. The results were obtained using the synthetic aperture algorithm with transmit and receive signals correction based on a single element directivity function.

Endothelial Specificity of ICAM2, Flt-1, and Tie2 Promoters In Vitro and In Vivo

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.

Optical Coherence Tomography Combined with the Confocal Microscopy Method and Fluorescence for Class V Cavities Investigations

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.

Effects of a Methanol Fraction of the Leaves of Leonotis leonurus on the Blood Pressure and Heart Rate of Normotensive Male Wistar Rats

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.

Comparison of Classical and Ultrasound-Assisted Extractions of Hyphaene thebaica Fruit and Evaluation of Its Extract as Antibacterial Activity in Reducing Severity of Erwinia carotovora

Erwinia carotovora var. carotovora is the main cause of soft rot in potatoes. Hyphaene thebaica was studied for biocontrol of E. carotovora which inhibited growth of E. carotovora on solid medium, a comparative study of classical and ultrasound-assisted extractions of Hyphaene thebaica fruit. The use of ultrasound decreased significant the total time of treatment and increase the total amount of crude extract. The crude extract was subjected to determine the in vitro, by a bioassay technique revealed that the treatment of paper disks with ultrasound extraction of Hyphaene thebaica reduced the growth of pathogen and produced inhibition zones up to 38mm in diameter. The antioxidant activity of ultrasound-ethanolic extract of Doum fruits (Hyphaene thebaica) was determined. Data obtained showed that the extract contains the secondary metabolites such as Tannins, Saponin, Flavonoids, Phenols, Steroids, Terpenoids, Glycosides and Alkaloids.

Biosynthesis and In vitro Studies of Silver Bionanoparticles Synthesized from Aspergillusspecies and its Antimicrobial Activity against Multi Drug Resistant Clinical Isolates

Antimicrobial resistant is becoming a major factor in virtually all hospital acquired infection may soon untreatable is a serious public health problem. These concerns have led to major research effort to discover alternative strategies for the treatment of bacterial infection. Nanobiotehnology is an upcoming and fast developing field with potential application for human welfare. An important area of nanotechnology for development of reliable and environmental friendly process for synthesis of nanoscale particles through biological systems In the present studies are reported on the use of fungal strain Aspergillus species for the extracellular synthesis of bionanoparticles from 1 mM silver nitrate (AgNO3) solution. The report would be focused on the synthesis of metallic bionanoparticles of silver using a reduction of aqueous Ag+ ion with the culture supernatants of Microorganisms. The bio-reduction of the Ag+ ions in the solution would be monitored in the aqueous component and the spectrum of the solution would measure through UV-visible spectrophotometer The bionanoscale particles were further characterized by Atomic Force Microscopy (AFM), Fourier Transform Infrared Spectroscopy (FTIR) and Thin layer chromatography. The synthesized bionanoscale particle showed a maximum absorption at 385 nm in the visible region. Atomic Force Microscopy investigation of silver bionanoparticles identified that they ranged in the size of 250 nm - 680 nm; the work analyzed the antimicrobial efficacy of the silver bionanoparticles against various multi drug resistant clinical isolates. The present Study would be emphasizing on the applicability to synthesize the metallic nanostructures and to understand the biochemical and molecular mechanism of nanoparticles formation by the cell filtrate in order to achieve better control over size and polydispersity of the nanoparticles. This would help to develop nanomedicine against various multi drug resistant human pathogens.