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Preparation Techniques (preparation + techniques)

Distribution by Scientific Domains

Medical Sciences	50%
Chemistry	28%
Life Sciences	17%

Kinds of Preparation Techniques

sample preparation techniques

Selected Abstracts

Preparation Techniques for the Injection of Human Autologous Cartilage: An Ex Vivo Feasibility Study,

THE LARYNGOSCOPE, Issue 1 2008
J Pieter Noordzij MD
Abstract Objectives: To determine the optimum donor site and preparation technique for injecting human autologous cartilage as a potentially permanent implant material for vocal fold medialization. Study Design: Prospective ex vivo experimental model. Methods: Human nasal septal and auricular cartilage was obtained from eight surgical cases after institutional review board approval. The auricle and nasal septum were chosen as potential donor sites because of ease of accessibility, volume of cartilage potentially available, and minimal subsequent cosmetic deformity after the tissue harvesting procedure. Various preparation techniques readily available in most operating rooms were tested for their efficacy in generating an injectable cartilage slurry. The various cartilage slurries were injected through sequentially smaller needles and examined cytologically. Results: The best injection properties for both nasal septal and auricular cartilage were obtained by drilling the cartilage down with a 5 mm otologic cutting bur, which allowed free passage through an 18 gauge needle. Cytologic examination of drilled septal cartilage showed good uniformity of cartilage pieces with a mean largest dimension of 0.44 ± 0.33 mm, and 33% of lacunae contained viable-appearing chondrocytes. Cytologic examination of drilled auricular cartilage was similar, exceptonly 10% of lacunae were occupied by chondrocytes. Other techniques tested (knife, morselizer, and cartilage crusher) did not yield injectable cartilage slurries. Conclusions: Both nasal septal and auricular cartilage can be prepared for injection via an 18 gauge needle using a cutting otologic bur. Further testing of in vivo viability and long-term volume retention is needed. [source]

Update on Semen Technologies for Animal Breeding,

REPRODUCTION IN DOMESTIC ANIMALS, Issue 1 2006
JM Morrell
Contents Despite the scale of the livestock breeding industry, where many millions of insemination doses are prepared each year, sperm preparation techniques are used infrequently in animal assisted reproduction compared with its human counterpart. However, some of the techniques used for human sperm preparation, for example, density gradient centrifugation, improve the quality of sperm preparations which is, in turn, reflected by an increased conception rate. The preparation technique separates motile spermatozoa with normal morphology and intact DNA from the total sperm population, leaving behind immature or senescent spermatozoa, morphologically abnormal ones and those with damaged DNA. Furthermore, the motile spermatozoa are removed from the seminal plasma which carries cells, cellular debris and reactive oxygen species, as well as pathogens. Gradient-prepared spermatozoa survive longer, either in liquid storage or when cryopreserved, and are free of bacteria and viral infectivity if prepared carefully. Preparation techniques such as density gradient centrifugation, or the simplified single layer centrifugation technique, have considerable potential for aiding sperm preparation from poor quality semen samples, such as may be obtained from unselected semen donors in captive breeding programmes, or from performance horses. Moreover, the removal of pathogens has important implications, both for disease control and for avoiding the use of antibiotics in semen extenders, which can be detrimental to sperm survival. [source]

Osteological Development of the Garfish (Belone belone) Larvae

ANATOMIA, HISTOLOGIA, EMBRYOLOGIA, Issue 5 2009
S. Ku
Summary Garfish, Belone belone (Linnaeus, 1761) is an elongate, slander fish inhabiting the Eastern Atlantic and Mediterranean Sea. These fish from the Belonidae (Actinopterygii) family have important commercial value for Croatian fisheries. Samples for research were collected from experimental hatching in the Novigrad Sea. Preparation techniques included fixation in buffered formalin, trypsin clearing and staining with alcian blue and alizarin red. As little is known of garfish osteology and bone morphology, the main goal of this study was to describe ossification process in garfish fry. At hatching, no skeletal structure is present. Newly-hatched larvae also had no osteological elements. Ossification started at 7 day post-hatching (DPH) [total length (TL) 18 mm] with head bones and vertebral neural arch. Head skeleton continued to develop mostly over the period from 7 to 10 DPH. At 21 DPH (TL 49 mm), ossification process seemed to be finished, but it was not possible to distinguish borders of all bones. The primary interest of our research was to understand the growth dynamics as well as transformation of supporting body elements from cartilage to bone. At the end, developmental characteristics and functional aspects of this formation in different fish species are discussed. [source]

REACH-driven developments in analysis and physicochemistry,

FLAVOUR AND FRAGRANCE JOURNAL, Issue 3 2010
A. Chaintreau
Abstract The enforcement of the REACH regulation in the fragrance domain has created new challenges for the analytical and physical chemist. Many chemicals used as perfumery ingredients are hydrophobic, because low-polar compounds exhibit a higher substantivity (i.e. persistence after application) than do polar compounds. As a result, the usual protocols are often unsuitable and new methods must be developed. Biodegradation studies sometimes call for the quantification of traces of such hydrophobic analytes in complex media (e.g. waste water, aqueous surfactant solutions). Existing sample preparation techniques are either inefficient or time consuming. A new approach is proposed, based on single-use absorbants, which allows accurate quantification down to the 100 ppb range. This extremely simple technique allows good throughput analyses. Determining the environmental profile of a compound requires the determination of some physical constants. Among these, solubility in water can be obtained from theoretical models or experimentally, but the resulting values may greatly differ as a function of the model or the protocol. Several experimental approaches are critically discussed and compared with a reference technique. The air-to-water partition coefficients are determined by using an improved version of the previously developed static-and-trapped headspace technique. Copyright © 2009 John Wiley & Sons, Ltd. [source]

The Effect of the Anode Loading and Method of MEA Fabrication on DMFC Performance

FUEL CELLS, Issue 3 2007
T. V. Reshetenko
Abstract The influence of the Pt-Ru anode loading and MEA preparation techniques on direct methanol fuel cell (DMFC) performance is studied. Two different anode catalyst layer preparation techniques are employed. One is the direct coating of anode catalyst ink on a membrane to form a catalyst coated membrane, CCManode, and the other is the coating of the ink on the diffusion layers, which generates a catalyst coated substrate, CCSanode. The power density of a combined CCManode/CCScathode MEA is higher than for a CCSanode/CCScathode MEA. The main difference in the performance is observed in the high current density region, where two-phase flow is present and mass transfer processes govern the performance. The CCManode and CCSanode have different macroscopic structures, while showing the same microscopic morphology. Based on their morphological differences, it is expected that the combination of the CCManode and carbon paper provides the more homogeneous removal of CO2 at high currents. The authors suggest that the application of the CCManode with an optimal anode loading improves anode mass transfer, reduces methanol crossover, and enhances the electrochemical reactions. [source]

Histopathological profile of surgically removed persistent periapical radiolucent lesions of endodontic origin

INTERNATIONAL ENDODONTIC JOURNAL, Issue 3 2009
R. M. Love
Abstract Aim, To determine the types of periapical lesions associated with root filled teeth with persistent periapical pathosis that required surgical treatment based on specific inclusion and exclusion criteria. Methodology, Periapical lesions from a consecutive clinical sample of 100 patients were examined histopathologically to determine a definitive diagnosis. Results, Females were more represented (n = 56), the average age was 46.5 years and there were no age differences between gender or lesion type. A diagnosis of periapical granuloma was the most common finding with a similar number present in females (n = 40) and males (n = 37). A cyst was present in 18% of the cases with a majority of females (n = 12) represented in the sample. Evidence of foreign material, with an appearance consistent with endodontic sealer materials, was seen in 25 periapical granulomas, two cysts and one scar. Two periapical scars were seen, one had a history of apicectomy and amalgam root-end filling while the other was associated with extruded root filling material. Conclusions, By using defined clinical inclusion and exclusion criteria a predictable clinical diagnosis of a persistent periapical lesion due to endodontic origin can be reliably made. Periapical granulomas and cysts were the most common periapical lesions of endodontic origin associated with persistent periapical pathosis with the overall incidence of periapical cysts similar to previous studies. The presence of endodontic material in a high proportion of periapical lesions suggests a cause-effect association with the inference that clinicians should employ canal preparation techniques that limit apical extrusion of material. [source]

Dentine removal in the coronal portion of root canals following two preparation techniques

INTERNATIONAL ENDODONTIC JOURNAL, Issue 11 2007
G. Plotino
Abstract Aim, To measure the root canal area and the reduction of the mesial and buccal/lingual wall thickness at the level of the coronal interference in mesial roots of mandibular molars after instrumentation with a crown-down or a simultaneous root canal preparation technique. Methodology, Twenty mesial roots of first mandibular molars with a moderate root canal curvature were embedded in resin and sectioned horizontally at the level of the coronal interference, using a modification of the Bramante technique. After scanning and processing, the sections were reassembled. One root canal of each root was prepared using ProTaper instruments, while Mtwo instruments were used in the other root canal of the same mesial root. After scanning and processing, the data obtained were analysed for two parameters: changes in root canal area after instrumentation (,A) and reduction of the mesial and buccal/lingual wall thickness (,T). The data were subjected to Student's t -tests for statistical analysis at a significance level of P < 0.05. Results, No statistically significant differences were found between the two groups with respect to the changes in the areas (,A) at the level considered (P = 0.410). No statistically significant differences were noticed between the two groups for dentine thickness (,T) of both the mesial wall (P = 0.077) and the buccal or lingual wall (P = 0.171). Conclusions, There was no difference between the ProTaper and Mtwo groups for the amount of dentine removed. [source]

A comparison of spreader penetration depth and load required during lateral condensation in teeth prepared using various root canal preparation techniques

INTERNATIONAL ENDODONTIC JOURNAL, Issue 8 2005
S. F. Dulaimi
Abstract Aim, To compare the influence of various root canal preparation techniques on spreader penetration depth and load required during lateral condensation with gutta-percha and sealer. Methodology, Eighty extracted human teeth with single and straight canals were used. Twenty teeth were instrumented using one of four root canal preparation techniques. The four preparation techniques were: step-back technique without Gates-Glidden drills, step-back technique with Gates-Glidden drills, crown-down pressureless technique and hybrid technique (step-down/step-back). After root canal preparation had been completed a simulated periodontal ligament was fabricated from a uniform layer of silicone impression material. The roots were then mounted in an acrylic resin to simulate the physical condition found in tooth socket. A standardized stainless steel hand spreader of the same size as the master apical file was mounted in an Instron testing machine and lateral compaction with gutta-percha and sealer was performed. The load value was recorded from the Instron testing machine. The spreader penetration depths were measured with an endodontic ruler. The data obtained were analysed statistically using anova and Student's t -tests. Results, No significant difference in initial spreader load needed to condense the master cone was found amongst the four canal preparation techniques (P > 0.05). The step-back technique with Gates-Glidden drills and the hybrid technique demonstrated the least difference between the initial spreader penetration and the working length (mean 1.925 and 2.25 mm, respectively). The step-back technique without Gates-Glidden drills and the crown-down pressureless technique had the greatest difference between initial spreader penetration and the working length (mean 4.425 and 4.75 mm, respectively). Conclusion, The flare created by canal preparation affected spreader penetration depth, but had no effect on the spreader load. [source]

Effects of rotary instruments and ultrasonic irrigation on debris and smear layer scores: a scanning electron microscopic study

INTERNATIONAL ENDODONTIC JOURNAL, Issue 7 2002
B. E. Mayer
Abstract Aim This study evaluated debris and smear layer scores after two types of instruments manufactured from different alloys were used to ultrasonically activate irrigants during canal preparation. The influence of two rotary preparation techniques on cleanliness of the shaped canals was also studied. Methodology Apical stops were prepared to size 45 in 42 single-canalled extracted premolars and canines, which were divided into six equal groups. Groups 1, 2 and 3 were prepared by ProFile .04 (PF) while groups 4, 5 and 6 were prepared by Lightspeed (LS). All groups were irrigated using 5.25% NaOCl and 17% EDTA. Irrigants in groups 2 and 5 were ultrasonically activated using a size 15 steel K-file and by a blunt flexible nickel,titanium wire in groups 3 and 6. Groups 1 and 4 served as negative controls. Roots were split and canal walls examined at 15×, 200× and 400× magnification in an SEM. Smear layer and debris scores were recorded at 3, 6 and 9 mm levels using a 5-step scoring scale and a 200-µm grid. Means were tested for significance using nonparametric Mann,Whitney U and Kruskal,Wallis tests. Results Debris and smear layer scores for the six groups varied from 1.98 ± 1.04 to 3.47 ± 0.97 and from 1.37 ± 0.4 to 2.36 ± 0.99, respectively. Although all groups had significantly higher smear layer and debris scores at the 3 mm levels compared to the 9 mm levels (P < 0.05), no significant differences were recorded due to the ultrasonic energy transmitted by the two alloys. Conclusion Ultrasonically activated irrigants did not reduce debris or smear layer scores. This finding was not influenced by the material nor by the design of the instrument used to transmit ultrasonic activation. [source]

Outcomes of the International Union of Crystallography Commission on Powder Diffraction Round Robin on Quantitative Phase Analysis: samples 2, 3, 4, synthetic bauxite, natural granodiorite and pharmaceuticals

JOURNAL OF APPLIED CRYSTALLOGRAPHY, Issue 4 2002
Nicola V. Y. Scarlett
The International Union of Crystallography (IUCr) Commission on Powder Diffraction (CPD) has sponsored a round robin on the determination of quantitative phase abundance from diffraction data. The aims of the round robin have been detailed by Madsen et al. [J. Appl. Cryst. (2001), 34, 409,426]. In summary, they were (i) to document the methods and strategies commonly employed in quantitative phases analysis (QPA), especially those involving powder diffraction, (ii) to assess levels of accuracy, precision and lower limits of detection, (iii) to identify specific problem areas and develop practical solutions, (iv) to formulate recommended procedures for QPA using diffraction data, and (v) to create a standard set of samples for future reference. The first paper (Madsen et al., 2001) covered the results of sample 1 (a simple three-phase mixture of corundum, fluorite and zincite). The remaining samples used in the round robin covered a wide range of analytical complexity, and presented a series of different problems to the analysts. These problems included preferred orientation (sample 2), the analysis of amorphous content (sample 3), microabsorption (sample 4), complex synthetic and natural mineral suites, along with pharmaceutical mixtures with and without an amorphous component. This paper forms the second part of the round-robin study and reports the results of samples 2 (corundum, fluorite, zincite, brucite), 3 (corundum, fluorite, zincite, silica flour) and 4 (corundum, magnetite, zircon), synthetic bauxite, natural granodiorite and the synthetic pharmaceutical mixtures (mannitol, nizatidine, valine, sucrose, starch). The outcomes of this second part of the round robin support the findings of the initial study. The presence of increased analytical problems within these samples has only served to exacerbate the difficulties experienced by many operators with the sample 1 suite. The major difficulties are caused by lack of operator expertise, which becomes more apparent with these more complex samples. Some of these samples also introduced the requirement for skill and judgement in sample preparation techniques. This second part of the round robin concluded that the greatest physical obstacle to accurate QPA for X-ray based methods is the presence of absorption contrast between phases (microabsorption), which may prove to be insurmountable in some circumstances. [source]

Synthesis of bulk MgB2 superconductors by pulsed electric current

AICHE JOURNAL, Issue 7 2006
A. M. Locci
Abstract A preparation method to simultaneously synthesize and consolidate bulk MgB2 superconductors from Mg and B commercial elemental powders by means of the spark plasma sintering technique is reported. The influence of process parameters on sintering process dynamics as well as product characteristics, determined by transport and magnetic measurements, is investigated. The superconducting properties of the obtained samples, and particularly the critical current density, are comparable or better than those corresponding to other MgB2 preparation techniques. Thus, the superconductive properties of the bulk MgB2 materials synthesized in this work are suitable for selected applications, such as magnetic levitation, magnetic screening, and fault current limiters. It should be finally noted that the proposed method represents a particularly rapid preparation route as compared to other techniques. © 2006 American Institute of Chemical Engineers AIChE J, 2006 [source]

An in vitro assessment of the strength of porcelain veneers dependent on tooth preparation

JOURNAL OF ORAL REHABILITATION, Issue 12 2000
P. Hahn
The treatment of teeth using veneer restorations combines aesthetic and functional improvements with a low destructive technique. Different kinds of tooth preparation techniques are described in the literature for this kind of restoration. This in vitro study aimed to examine the influence of the incisal preparation on the loadability of teeth restored with porcelain laminate veneers. Thirty-six selected mandibular incisors were randomly assigned to three groups with 12 teeth per group. In the first group, only the facial surface was prepared. In the second group, the preparation included a rounded incisal edge and a distinct chamfer lingually. The third group served as an unprepared control. Empress® veneers were then fabricated and cemented with a low viscous luting composite material. After 120 days storage in Ringer's solution, the specimens were loaded incisally to the point of failure. Statistical analysis of the results showed significant differences between the series (P=0·0103). Group 2 (with preparation of the incisal edge) exhibited the lowest fracture resistance (466±99 N) (N, mean forces). When prepared only facially, the teeth restored with Empress® veneers reached the strength of unprepared teeth. Compared with the biting force described for incisors in the literature, the in vitro loadability reached in this investigation seems to jusitify the clinical use of both preparation designs tested. [source]

A new single grating spectrograph for ultraviolet Raman scattering studies

JOURNAL OF RAMAN SPECTROSCOPY, Issue 5 2006
Lutz Hecht
Abstract A state-of-the-art single grating spectrograph for Raman scattering studies within the deep ultraviolet (DUV) region of the electromagnetic spectrum is discussed. It is based on a high throughput DUV version of a single-stage monochromator originally designed for use in the visible spectral region. Its key components are two identical, newly designed calcium fluoride camera lenses each consisting of five different individual optical elements. The first of these lenses collimates the Raman scattered DUV radiation entering the spectrometer through its entrance slit. The second lens focuses the collimated beam of dispersed Raman scattered DUV radiation emerging from a high-resolution reflection grating onto a charge coupled device (CCD) detector with enhanced DUV sensitivity. A novel high transmission edge filter is used as a blocking device for a sufficient rejection of the Rayleigh line generating a relatively sharp transmittance cutoff at a Stokes Raman wavenumber shift of about ,450 cm,1 employing 257 nm DUV excitation. Overall, this new spectrograph enables rapid collection of Stokes DUV Raman scattered photons at f/2 wide apertures with sufficiently large signal-to-noise ratios (SNRs) in relatively short acquisition times and with an effective spectral resolution of approximately ,6 cm,1. Backscattered Raman spectra of the following chemicals are presented as typical results illustrating the excellent performance characteristics of this new DUV spectrograph for a variety of experimental conditions within different scattering scenarios and for a relatively wide range of commonly used sample preparation techniques: neat cyclohexane, laboratory air, polycrystalline D -glucose, single crystal L -alanine and a dilute aqueous solution of 2,-deoxyadenosine. Copyright © 2005 John Wiley & Sons, Ltd. [source]

ANACORACID SHARKS FROM THE ALBIAN (LOWER CRETACEOUS) PAWPAW SHALE OF TEXAS

PALAEONTOLOGY, Issue 4 2007
MIKAEL SIVERSON
Abstract:, Recent collecting from the Pawpaw Shale in north-east Texas has yielded several hundred teeth of anacoracid sharks. The material allows for a much-needed revision of the Late Albian anacoracids from North America. The previously recognized Squalicorax sp., also referred to as S. volgensis in more recent publications, is a mix of two different species: S. priscoserratus sp. nov. and S. pawpawensis sp. nov. In addition to these two new species, a single tooth is assigned to S. aff. S. baharijensis. Our data indicate that anacoracids were a considerably more diverse group in the North American Cretaceous than previously thought. We attribute much of the underestimation of diversity to vague species concepts, poor preparation techniques and the associated lack of attention to certain dental features, in particular neck morphology, root surface porosity and the root's vascularization. [source]

Improvements for the visualization of low-molecular weight protein and peptides of human tears using MALDI

PROTEINS: STRUCTURE, FUNCTION AND BIOINFORMATICS, Issue 17 2008
Terry Nguyen-Khuong
Abstract Many low-molecular weight proteins and peptides in human tears are potentially bioactive proteins but the range and number of these is yet to be fully characterized. A number of different sample preparation techniques were used to maximize the visualization of peptides from reflex tears. Samples were pretreated using precipitation and filtration techniques prior to analyses using MALDI-TOF MS. Peptides were searched for between 700 to 4000 m/z. Sample dilution in several different buffer systems followed by filtration with a 30-kDa cutoff filter and C18 reverse phase microcolumn purification produced significantly (p = 0.049) more peaks in tears than other methods used to prepare tears prior to MALDI-TOF MS. This study has established a technique for optimizing the visualization of naturally occurring peptides in tears. [source]

Matrix vapor deposition/recrystallization and dedicated spray preparation for high-resolution scanning microprobe matrix-assisted laser desorption/ionization imaging mass spectrometry (SMALDI-MS) of tissue and single cells

RAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 3 2010
Werner Bouschen
Matrix preparation techniques such as air spraying or vapor deposition were investigated with respect to lateral migration, integration of analyte into matrix crystals and achievable lateral resolution for the purpose of high-resolution biological imaging. The accessible mass range was found to be beyond 5000 u with sufficient analytical sensitivity. Gas-assisted spraying methods (using oxygen-free gases) provide a good compromise between crystal integration of analyte and analyte migration within the sample. Controlling preparational parameters with this method, however, is difficult. Separation of the preparation procedure into two steps, instead, leads to an improved control of migration and incorporation. The first step is a dry vapor deposition of matrix onto the investigated sample. In a second step, incorporation of analyte into the matrix crystal is enhanced by a controlled recrystallization of matrix in a saturated water atmosphere. With this latter method an effective analytical resolution of 2,µm in the x and y direction was achieved for scanning microprobe matrix-assisted laser desorption/ionization imaging mass spectrometry (SMALDI-MS). Cultured A-498 cells of human renal carcinoma were successfully investigated by high-resolution MALDI imaging using the new preparation techniques. Copyright © 2010 John Wiley & Sons, Ltd. [source]

Update on Semen Technologies for Animal Breeding,

Revegetation Methods for High-Elevation Roadsides at Bryce Canyon National Park, Utah

RESTORATION ECOLOGY, Issue 2 2004
S. L. Petersen
Abstract Establishment of native plant populations on disturbed roadsides was investigated at Bryce Canyon National Park (BCNP) in relation to several revegetation and seedbed preparation techniques. In 1994, the BCNP Rim Road (2,683,2,770 m elevation) was reconstructed resulting in a 23.8-ha roadside disturbance. Revegetation comparisons included the influence of fertilizer on plant establishment and development, the success of indigenous versus commercial seed, seedling response to microsites, methods of erosion control, and shrub transplant growth and survival. Plant density, cover, and biomass were measured 1, 2, and 4 years after revegetation implementation (1995,1998). Seeded native grass cover and density were the highest on plots fertilized with nitrogen and phosphorus, but by the fourth growing season, differences between fertilized and unfertilized plots were minimal. Fertilizers may facilitate more rapid establishment of seeded grasses following disturbance, increasing soil cover and soil stability on steep and unstable slopes. However the benefit of increased soil nutrients favored few of the desired species resulting in lower species richness over time compared to unfertilized sites. Elymus trachycaulus (slender wheatgrass) plants raised from indigenous seed had higher density and cover than those from a commercial seed source 2 and 4 years after sowing. Indigenous materials may exhibit slow establishment immediately following seeding, but they will likely persist during extreme climatic conditions such as cold temperatures and relatively short growing seasons. Seeded grasses established better near stones and logs than on adjacent open microsites, suggesting that a roughened seedbed created before seeding can significantly enhance plant establishment. After two growing seasons, total grass cover between various erosion-control treatments was similar indicating that a variety of erosion reduction techniques can be utilized to reduce erosion. Finally shrub transplants showed minimal differential response to fertilizers, water-absorbing gels, and soil type. Simply planting and watering transplants was sufficient for the greatest plant survival and growth. [source]

Preparation Techniques for the Injection of Human Autologous Cartilage: An Ex Vivo Feasibility Study,

Effects of H2O2 exposure on human sperm motility parameters, reactive oxygen species levels and nitric oxide levels

ANDROLOGIA, Issue 3 2010
S. S. Du Plessis
Summary Research has revealed that reactive oxygen species (ROS) negatively affect sperm function, both in vivo and in vitro. Sperm preparation techniques for assisted reproductive technologies (ART) are potential causes for additional ROS production. This study aimed to correlate the concentration of exogenous H2O2 with sperm motility parameters and intracellular ROS and nitric oxide (NO) levels to reiterate the importance of minimising ROS levels in ART. Human spermatozoa from 10 donors were incubated and exposed to different exogenous H2O2 concentrations (0, 2.5, 7.5 and 15 ,m). Subsequently, motility was determined using computer-aided semen analysis, while ROS (2,7-dichlorofluorescin diacetate) and NO (diaminofluorescein-2/diacetate) were analysed using fluorescence-activated cell sorting. Results showed that H2O2 did affect the sperm parameters. Exogenous H2O2 was detrimental to motility and resulted in a significant increase in overall ROS and NO levels. A significant increase in static cells was seen as well. It is important to elucidate the mechanisms between intracellular ROS levels with sperm motility parameters. While this experiment demonstrated a need to reduce exogenous ROS levels during ART, it did not illustrate the cause and effect relationship of intracellular ROS and NO levels with sperm motility. Further research needs to be conducted to define a pathological level of ROS. [source]

The in vitro effects of melatonin on human sperm function and its scavenging activities on NO and ROS

ANDROLOGIA, Issue 2 2010
S. S. Du Plessis
Summary Various systems of antioxidants exist endogenously in the body to help protect it against free radical damage by scavenging excessive ROS and RNS. Melatonin, a hormone secreted by the pineal gland, and responsible for controlling the circadian rhythm, is one such endogenous antioxidant. Melatonin has been reported to be present in human seminal fluid, but its antioxidant activities in semen are rather contradictory. This study aimed at establishing the effects of melatonin treatment on human spermatozoa. Spermatozoa were incubated with 2 mm melatonin (120 min, 37 °C, 5% CO2) after which motility parameters were measured by computer aided motility analysis, while cell viability (PI), intracellular NO (DAF-2/DA) and ROS (DCFH-DA) were assessed using flow cytometry. In vitro melatonin treated samples (n = 12) showed a significantly higher percentage of motile, progressive motile and rapid cells, while simultaneously reducing the number of nonviable spermatozoa when compared with the control. Endogenous NO was significantly decreased, but no effect was observed on ROS levels. From these results, it can be concluded that melatonin was able to directly or indirectly scavenge NO, as indicated by the reduction in 4,5-diaminofluorescein-2/diacetate fluorescence. Future studies will indicate whether melatonin treatment during sperm preparation techniques could protect spermatozoa from excessive NO production. [source]

Limitation of immunoaffinity column for the removal of abundant proteins from plasma in quantitative plasma proteomics

BIOMEDICAL CHROMATOGRAPHY, Issue 5 2009
Tomoko Ichibangase
Abstract In plasma proteomics, before a proteome analysis, it is essential to prepare protein samples without high-abundance proteins, including albumin, via specific preparation techniques, such as immunoaffinity capture. However, our preliminary experiments suggested that functional changes with use alter the ability of the immunoaffinity column. Thus, in this study, to evaluate the changes of the removal ability of abundant proteins from plasma by the immunoaffinity column, plasma proteome analysis was performed for the long-term test for the reproducibility of the affinity column using the fluorogenic derivatization,liquid chromatography,tandem mass spectrometry method combined with an IgY column. The specific adsorption for albumin decreased with an increase in the number of the column usage before its expiration date. Moreover, it was demonstrated that hydrophobic high molecular weight compounds in plasma adsorbed onto the column materials surface contributed to the functional changes from specific immunoaffinity adsorption into hydrophobic interaction. These results suggested that, in quantitative plasma proteomics studies, it is important to keep in mind the risk of not only the nonselective loss but also the changes in the adsorption ability of the immunoafinity column. Copyright © 2008 John Wiley & Sons, Ltd. [source]

Progress technology in microencapsulation methods for cell therapy

BIOTECHNOLOGY PROGRESS, Issue 4 2009
Jean-Michel Rabanel
Abstract Cell encapsulation in microcapsules allows the in situ delivery of secreted proteins to treat different pathological conditions. Spherical microcapsules offer optimal surface-to-volume ratio for protein and nutrient diffusion, and thus, cell viability. This technology permits cell survival along with protein secretion activity upon appropriate host stimuli without the deleterious effects of immunosuppressant drugs. Microcapsules can be classified in 3 categories: matrix-core/shell microcapsules, liquid-core/shell microcapsules, and cells-core/shell microcapsules (or conformal coating). Many preparation techniques using natural or synthetic polymers as well as inorganic compounds have been reported. Matrix-core/shell microcapsules in which cells are hydrogel-embedded, exemplified by alginates capsule, is by far the most studied method. Numerous refinement of the technique have been proposed over the years such as better material characterization and purification, improvements in microbead generation methods, and new microbeads coating techniques. Other approaches, based on liquid-core capsules showed improved protein production and increased cell survival. But aside those more traditional techniques, new techniques are emerging in response to shortcomings of existing methods. More recently, direct cell aggregate coating have been proposed to minimize membrane thickness and implants size. Microcapsule performances are largely dictated by the physicochemical properties of the materials and the preparation techniques employed. Despite numerous promising pre-clinical results, at the present time each methods proposed need further improvements before reaching the clinical phase. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009 [source]

Microbial Surfaces Investigated Using Atomic Force Microscopy

BIOTECHNOLOGY PROGRESS, Issue 6 2004
Anastassia V. Bolshakova
This paper is dedicated to atomic force microscopy (AFM) as a progressive tool for imaging bacterial surfaces and probing their properties. The description of the technique is complemented by the explanation of the methodapos;s artifacts typical, in particular, for the imaging of bacterial cells. Sample preparation techniques are summarized in a separate section. Special attention is paid to the differences in imaging of Gram-positive and Gram-negative bacteria. Probing of mechanical properties, including elastic modulus, fragility, and adhesion of the cell walls is emphasized. The advantages of AFM in the studies of real-time cellular dynamical processes are illustrated by the experiment with the germination of spores. [source]

The effect of three different calcium phosphate implant coatings on bone deposition and coating resorption: a long-term histological study in sheep

CLINICAL ORAL IMPLANTS RESEARCH, Issue 3 2005
Christian Schopper
Abstract: The present study investigated the hypothesis that hydroxyapatite (HA), tricalcium phosphate (TCP), and a HA-gel coated on endosseous titanium (Ti) implants by spark discharging (SD) and dip coating would achieve predictable osseointegration without evident bioresorption of the coatings on the long term. A costal sheep model was used for the implantation of the HA/SD, HA/TCP/SD, and HA-gel/SD specimens, which were retrieved 6 and 12 months following implantation. HA and Ti coatings on implants obtained by conventional plasma spraying (HA/PS, Ti/PS) were used as controls. Microscopy showed that osseointegration was achieved from all types of implants. No evidence for bioresorption of the HA/SD, HA/TCP/SD, and HA-gel/SD coatings was present but cohesive failure with disruption of the coating/implant interface was seen. A statistical analysis of the histomorphometrical data showed no time-dependent effect, however. HA/PS coatings achieved significantly higher bone,implant contact (BIC) percentages of the total implant surface (toBIC) than the other types of coatings (P=0.01). If the BIC percentages were traced separately for implant portions placed into cortical and cancellous bone (coBIC and caBIC, respectively), detailed analysis showed that the caBIC values of HA-gel/SD and HA/PS coatings were significantly higher than that of the other types of coatings (P=0.01). CaBIC values were highly correlated with toBIC values (P<0.001). The present study showed that the preparation techniques used produced thin, dense, and unresorbable coatings that achieved osseointegration. Compared with the control coatings, however, only HA-gel/SD coating can be recommended from the investigated preparation techniques for a future clinical use if a better coating cohesion is achieved. Résumé L'étude présente a étudié l'hypothèse que le recouvrement par de l'hydroxyapatite, du phosphate tricalcique et un gel d'hydroxyapatite sur les implants en titane par décharges spark et recouvrement par trempage pourrait apporter une ostéïntégration prévisible sans biorésorption importante des recouvrements à long terme. Un modèle de mouton a été utilisé pour l'implantation de spécimens HA/SD, HA/TCP/SD et gel-HA/SD qui ont été enlevés six et douze mois après leur insertion. Les implants recouverts d'hydroxyapatite et de titane obtenus par plasma-spray conventionnel (HA/PS et Ti/PS) ont été utilisés comme contrôles. La microscopie a montré que l'ostéoïntégration a été réalisée pour tous les types d'implants. Aucune évidence pour la biorésorption de HA/SD, HATCP/SD, et gel-HA/SD n'était présente mais un échec de cohésion avec destruction de l'interface implant/recouvrement a été mis en évidence. Une analyse statistique des données histomorphométriques ne montrait cependant aucun effet dépendant du temps. Les recouvrements HA/PS montraient des pourcentages de contact os/implant significativement plus importants de la surface implantaire totale (BIC) que les autres types de recouvrement (p=0,01). Lorsque les pourcentages de contact os-implant étaient lus séparément pour les portions implantaires placées dans l'os cortical ou l'os spongieux (respectivement coBIC et caBIC), l'analyse détaillée montrait que les valeurs caBIC du gel- HA/SD et des recouvrements HA/PS étaient significativement plus importants que dans les autres types de recouvrement (p<0,01). Les valeurs CaBIC étaient en relation étroite avec les valeurs toBIC (p<0,001). L'étude présente a montré que les techniques de préparation utilisées produisaient des recouvrements non-résorbables denses et fins qui permettaient l'ostéoïntégration. Cependant, comparé aux recouvrements contrôles, seul le recouvrement gel-HA/SD pouvait être recommandé avec les techniques de préparation étudiées pour une utilisation clinique future si une cohésion de recouvrement meilleure est assurée. Zusammenfassung Die vorliegende Studie untersuchte die Hypothese, dass Hydroxyapatit, Trikalziumphoshat und ein Hydroxyapatit-Gel als Beschichtung auf enossalen Ti-Implantaten zur voraussagbaren Osseointegration über einen langen Zeitraum ohne Bioresorption der Beschichtung führen. Die Beschichtungen wurden durch Funkenentladung und Tauchbeschichtung aufgetragen. Für die Implantation der HA/SD, HA/TCP/SD und HA-Gel/SD wurde ein Schafmodell verwendet. Die Proben wurden 6 und 12 Monate nach Implantation entnommen. Als Kontrolle dienten Hydroxyapatit- und Titanbeschichtungen (HA/PS und Ti/PS), welche mittels Plasmaspray aufgetragen worden waren. Die mikroskopische Untersuchung zeigte, das bei allen Implantattypen eine Osseointegration erreicht wurde. Bei den HA/SD, HA/TCP und HA-Gel/SD Beschichtungen waren keine Anzeichen von Bioresorption vorhanden, aber es konnten kohäsive Misserfolge mit Abrissen im Bereich der Implantat/Beschichtung-Berührungsfläche gesehen werden. Eine statistische Analyse der histomorphometrischen Daten zeigte jedoch keinen zeitabhängigen Effekt. Die HA/PS Beschichtungen erreichten signifikant höhere Knochen-Implantat-Kontakt Prozentwerte der gesamten Implantatoberfläche (toBIC) als die anderen Beschichtungen (P=0.01). Wenn die Knochen-Implantat-Kontakt Prozentwerte für Implantatbereiche, welche im kortikalen und spongiösen Knochen (coBIC und caBIC) lagen, separat ausgemessen wurden, so zeigte die detaillierte Analyse, dass die caBIC Werte von HA-Gel/SD und HA/PS Beschichtungen signifikant höher waren als bei allen anderen Typen von Beschichtungen (P=0.01). Die caBIC Werte zeigten eine starke Korrelation mit den toBIC Werten (P<0.001). Die Studie zeigte, dass das verwendete Herstellungsverfahren dünne, dichte und nicht resorbierbare Beschichtungen ergab, welche eine Osseointegration erreichten. Im Vergleich mit den Kontrollbeschichtungen können jedoch nur die HA-Gel/SD Beschichtungen der untersuchten Herstellungsverfahren für den weiteren klinischen Gebrauch empfohlen werden, falls eine bessere Kohäsion der Beschichtung erreicht werden kann. Resumen El presente estudio investigó la hipótesis de que la hidroxiapatita, el fosfato tricálcico y un gel de hidroxiapatita cubriendo implantes endoóseos de Ti por medio de chisporroteo e inmersión pueden lograr una osteointegración predecible sin una biorreabsorción evidente de las cubiertas a largo plazo. Se usó un modelo costal de oveja para la implantación de especímenes HA/SD, HA/TCP/SD, y gel-HA/SD que se retiraron a los 6 y a los 12 meses de la implantación. Como control se usaron cubiertas de hidroxiapatita y titanio en implantes obtenidos por medio de pulverización de plasma convencional (HA/SD, Ti/PS). La microscopía demostró que la osteointegración se logró en todos los tipos de implantes. No existió evidencia de biorreabsorción de las cubiertas HA/SD, HA/TCP/SD, y gel-HA/SD pero se observó fallos en la cohesión con disrupción de la interfase cubierta/implante. Un análisis estadístico de los datos histomorfométricos no mostró, sin embargo efectos dependientes del tiempo. Las cubiertas HA/PS lograron unos porcentajes de contacto hueso-implante significativamente mayores del total de la superficie del implante (toBIC) que los otros tipos de cubiertas (P=0.01). Si se ubicaran los porcentajes de contacto hueso-implante separadamente para porciones situadas dentro de hueso cortical o esponjoso (coBIC y caBIC respectivamente), un análisis detallado mostró que los valores caBIC de las cubiertas de HA-gel/SD y HA/PS fueron significativamente mayores que aquellos de los otros tipos de cubiertas (P<0.001). El presente estudio mostró que las técnicas de preparación usadas produjeron cubiertas finas, densas y no reabsorbibles que alcanzaron la osteointegración. De todos modos, comparadas con las cubiertas de control, solo la cubierta HA-gel/SD pude ser recomendada desde las técnicas de preparación investigadas para un futuro uso clínico si se lograse una mejor cohesión de cubierta. [source]