A survey among Flemish gastroenterologists about endoscopic sedation practices in colorectal cancer screening.

Background & study aims: The sedation levels and methods used for colonoscopy in colorectal cancer screening programs vary from country to country and from continent to continent. Little is known in the literature about how frequently the different sedation levels are used in colorectal cancer screening colonoscopies. We made a survey among all Flemish gastroenterologists (GI) to determine how frequently they use the different sedation modalities in this target population and to determine the motives of the GI to opt for one or another sedation modality. Patient and methods: An online survey was sent to all 329 Flemish GI by e-mail. A reminder e-mail was sent one month later. Participants could indicate how frequently (by percentage) they used the different sedation methods (no sedation, minimal sedation, conscious sedation, deep sedation) and which sedative medication they administered. In addition, they were asked to indicate their main motives for choosing a specific sedation method. Descriptive statistics were used. Results: 112 out of 329 GI answered the questionnaire (response rate 34%). Anesthesia monitored care is the most frequently used sedation modality, followed by conscious sedation. Patient preference is the main motive for most GI to use each sedation modality. Conclusions: Anesthesia monitored care is currently the most frequently used sedation regimen to perform a colonoscopy in the FIT positive population or in the colorectal cancer screening program in Flanders. The motives given by the GI for choosing one or another sedation modality are not always congruent with current scientific evidence or guidelines.

Characterization of porous silicon integrated in liquid chromatography chips.

Properties of porous silicon which are relevant for use of the material as a stationary phase in liquid chromatography chips, like porosity, pore size and specific surface area, were determined with high-resolution SEM and N(2) adsorption-desorption isotherms. For the anodization conditions investigated, porosity is between 20 and 60%, pore sizes between 2 and 5 nm and specific surface area between 130 and 410 m(2)/cm(3). It was established that under identical anodization conditions, porous layer formation is 10-15% slower on micromachined pillars than on flat substrates, and depends on geometrical parameters like pillar diameter and height and interpillar spacing. In microchannels containing pillars with a porous silicon shell, chromatographic experiments on a coumarin dye mixture were performed, which in comparison with non-porous pillars showed a significant increase of the retention factors, resulting from the large internal surface of the porous pillars. The increased relative retention of one of the coumarin dyes, C480, could be correlated quantitatively with the measured internal surface of the porous layer. Due to the small pore size, these porous shell columns are particularly suitable for analytical or preparative separation of low-molecular weight molecules, with applications in metabolomics, food quality control, or medical diagnostics.

Errors involved in the existing B-term expressions for the longitudinal diffusion in fully porous chromatographic media Part II: experimental data in packed columns and surface diffusion measurements.

Peak parking experiments have been performed on three RP-HPLC different columns, using two different components and a variable mobile phase composition. The aim of the study was to investigate whether the B-term diffusion expressions currently used in the literature (which are all Knox-type models) should be replaced by the effective diffusion expressions that have been developed in the frame of the effective medium theory (EMT). Although the EMT-expressions are not fully accurate either (the mathematics of the complex interactions between different diffusion zones that are in close contact are too demanding to catch them in an exact analytical expression), they at least are physically sound and do not violate Maxwell's basic law of diffusion. Further they also provide a much better approximation of the numerically calculated effective diffusivity in the theoretical test situation considered in part I. The present study shows that the values of the surface or stationary phase diffusion coefficient that are derived from peak parking models can depend heavily on the employed B-term model. The EMT-based B-term expressions lead to values of the surface diffusion coefficient that vary much less strongly with the phase retention factor than if one of the Knox-type models is used to analyze the data. This implies that, since all peak parking experiments that have been performed in the past have all been interpreted with a Knox-type model, the conclusions that have been drawn from these studies should all be moderated or at least revisited.

Errors involved in the existing B-term expressions for the longitudinal diffusion in fully porous chromatographic media Part I: computational data in ordered pillar arrays and effective medium theory.

Numerically solving the effective diffusion in a simplified representation of a chromatographic bed, it was found that the B-term expressions that have up to now been used in the literature, and which can all be reduced to either Deff=(gamma mDm+k'gamma sDs)/(1+k') or Deff=(gamma meDm+k''Dpart)/(1+k''), can no longer be considered to be unconditionally valid. This could be demonstrated by showing that the simulated diffusion data are in agreement with the mathematically sound effective medium theory (EMT), whereas the B-term expressions used up to now in literature are in conflict with the EMT, a theory that is widely accepted in all other fields of science. It is also shown that the use of the existing B-term expressions can lead to very large measurement errors (up to a 100% and more) for the determination of the stationary phase diffusion coefficient gamma sDs from peak parking experiments. The representation of the B-term diffusion should in the future hence be based on the Deff expressions that can be derived from the EMT. These are physically sound and are also more accurate than the classical B-term expressions used up to now.

Understanding and design of existing and future chromatographic support formats.

The present contribution reviews the use of alternative support formats as a means to surpass the chromatographic performance of the packed bed of spheres. First, a number of idealized structures are considered to obtain a general insight in how the performance of a chromatographic support depends on its shape and size, using the isocratic peak-capacity generation speed as the main performance indicator. Using this criterion, it is found that the packing density or, equivalently, the external porosity, is the most important of all geometrical shape factors. Depending on whether the sample consists of weakly or strongly retained components, the optimal external porosity can be expected to vary between 60% and a value near 100%. The optimal exploitation of a high external porosity, however, also requires overall shrinkage of the domain size, towards and into the sub-micron range. With the current fabrication technologies, this requirement seems difficult to achieve. In the presence of a lower limit on the characteristic support size, each range of desired plate numbers or peak capacities has its own optimal external porosity, ranging from a very low value (high packing density) for high speed, small peak capacity applications, to very high external porosities (low packing density) for applications requiring a very large peak capacity. Subsequently, the obtained theoretical insights are used to review and discuss the past and current research on alternative support formats. Finally, a number of emerging micro- and nano-fabrication technologies are introduced and their potential for the future production of supports with improved shape and homogeneity is discussed.

Use of the kinetic plot method to analyze commercial high-temperature liquid chromatography systems. I: Intrinsic performance comparison.

The present study aimed at mapping the separation speed potential of a critical pair on commercial high-temperature HPLC (HT-HPLC) supports at elevated temperatures. For this purpose, band broadening and pressure drop measurements were conducted on three different commercial HT-HPLC columns operated at various elevated temperatures but by keeping the same retention factor. The plate height data were subsequently transformed into a plot showing the minimal required analysis time needed to yield a given required effective plate number. For the considered RPLC alkylbenzene separations, it was found that the maximal gain in separation speed of the critical pair that can be obtained by varying the operating temperature from T=30 to 120 degrees C can be expected to be of the order of a factor of 3-4, if using an individually optimized column length for each considered temperature and if no secondary adsorption effects occur at the lower temperature. This gain factor, remaining more or less constant over the most relevant range of plate numbers, largely paralleled the reduction of the mobile phase viscosity accompanying the temperature increase.

On the 3-dimensional effects in etched chips for high performance liquid chromatography-separations.

In an attempt to quantify the potential of photolithographically etched micro-pillar arrays as a perfectly ordered alternative for the packed bed of spheres, the additional band broadening originating from the top and bottom plate has been investigated using computational fluid dynamics simulations. These calculations provide insight in the theoretical expectations that can be made for the experimental work that is currently being conducted by a number of groups. The calculations show that the additional band broadening contribution can be expected to go through a transient regime as a function of the axial distance along the array. In its fully developed regime and in the most relevant velocity range, the top and bottom wall contribution almost doubles the band broadening compared to the band broadening in a perfectly ordered array of non-porous, non-retentive pillars without top and bottom wall. Compared to the band broadening in an array of porous, retentive pillars on the other hand, the top and bottom wall-effect can be expected to become negligible. A simplified, phenomenological model yielding a first principles prediction of both the transient and the steady-state top and bottom wall band broadening as a function of the inter-pillar distance and the pillar height is proposed and shows good qualitative agreement with the exact calculations.

Use of the kinetic plot method to analyze commercial high-temperature liquid chromatography systems. II. Practically constrained performance comparison.

Using a set of experimentally determined plate height data obtained on three commercial high-temperature HPLC supports, and evaluating their isocratic separation speed potential under the application of a set of instrumental constraints, a qualitative map of the practically achievable critical pair separation speed potential of high-temperature HPLC has been established. The obtained data show that the gain in separation speed is more strongly affected by the instrumental limitations in the high-temperature range than it is for the low temperatures. For the presently considered case of alkylbenzene separations, the potential gain in analysis time that can be obtained by going from T=30 to 120 degrees C in the presence of a typical set of instrumental limitations nevertheless remains of the order of a factor of 2-4. The study also shows that improvements on the instrumentation side (increased detector frequency, pumping flow rate, smaller extra-column volumes, ...) are indispensable to fully benefit from the high temperature advantages for all separations requiring less than 10,000 effective theoretical plates.

Considerations for the use of ultra-high pressures in liquid chromatography for 2.1mm inner diameter columns.

The current contribution investigates the effects of viscous heat dissipation in chromatographic columns (with an emphasis on so-called narrow bore columns with an inner diameter of 2.1mm) using numerical simulations of the temperature and velocity profiles and the resulting band broadening, for the first time at operating pressures up to 2000bar. When operating columns under well-thermostatted conditions to maintain a constant temperature of the mobile phase, a dramatic increase in plate heights can be observed that voids any advantage one could expect from the possibility to use smaller particles offered by the increased pressure limit. It is also clearly demonstrated that, even when the column is not temperature controlled, the backflow of heat along the wall can causes a significant loss in performance under standard operating conditions in a still air oven. It is found that for operating pressure above 1250bar, a significant (relative to the typical column performance) contribution to the observed plate height will be caused by viscous heating effects, which increases with increasing temperature dependency of the retention factor. In addition, unprecedented experimental measurements of the temperature effects at an operating pressure up to 2600bar were performed on a 10cm long, 2.1mm ID column showing a dramatic temperature increase up to 60°C relative to the inlet temperature when using methanol as a mobile phase.

On the inhibitory action of cadmium on the donor side of photosystem II in isolated chloroplasts.

The inhibitory effect of the Cd2+ in the electron transport of the isolated chloroplasts has been observed by measuring the oxygen uptake from the solution and the fluorescence induction. Cd2+ is found to be an inhibitor on the donor side of Photosystem II and its action site, as determined by experiments using hydroxylamine and exogenous Mn, is supposed to be on the water-splitting enzyme itself. Moreover, physicochemical and physiological studies indicate that only the ionic form of Cd is acting at the level of the manganoprotein. It is not possible, from this work, to define precisely in which form Cd is taken up through the thylakoid membranes.

Influence of the packing heterogeneity on the performance of liquid chromatography supports.

We report on a series of plate height and flow resistance data obtained via computational fluid dynamics simulations in a simplified two-dimensional (2D) mimic of real packed bed and monolithic columns. By varying the external porosity (0.4 < epsilon < 0.8) and the degree of packing randomness, a good qualitative insight in the relationship between the packing porosity and heterogeneity and the general chromatographic performance parameters is obtained, unbiased by any differences in phase retention factor k', mobile phase diffusivity or viscosity or intra-skeleton porosity. The results provide a quantitative support for the use of domain size reduced plate heights as a means to compare the performance of chromatographic beds with a different porosity, as it was found that packings with a similar degree of packing heterogeneity yield very similar domain size reduced h(min)-values, nearly completely independent of the porosity. The study also clearly shows that the presence of preferential flow paths (inevitably accompanied by the presence of more clustered regions) leads to a decrease of the flow resistance, but also leads to a strong increase of the band broadening if supports with the same porosity epsilon and the same radial width are compared. For the presently considered 2D system, the flow resistance reduction is too small to overcome the corresponding strong increase in band broadening, such that the presence of preferential flow paths always leads to an overall increase of the separation impedance.

On the optimisation of the bed porosity and the particle shape of ordered chromatographic separation media.

We report on a theoretical study wherein we considered a large number of ordered two-dimensional porous pillar arrays with different pillar shapes and widely varying external porosity and calculated the flow resistance and the band broadening (under retentive conditions) over the complete range of practical velocities using a commercial computational fluid dynamics software package. It is found that the performance of the small porosity systems is very sensitive to the exact pillar shape, whereas this difference gradually disappears with increasing porosity. The obtained separation impedances are very small in comparison to packed bed and monolithic columns and decrease with increasing porosity. If accounting for the current micromachining limitations, a proper selection of the exact shape and porosity even becomes more critical, and different design rules are obtained depending on whether porous or non-porous pillars are considered.

Optimization and evaluation of radially interconnected versus bifurcating flow distributors using computational fluid dynamics modelling.

Two main groups of flow distributors, viz. "bifurcating distributors" (BF) and "radially interconnected distributors" (RI), as well as some hybrid distributors were investigated. Computational fluid dynamics was used to evaluate the performance of the distributors and to establish the design yielding the most uniform velocity field and the smallest variance for the bands emerging from the distributor. A minimum channel width of 100 µm was considered to allow the use of micro-milling techniques for chip fabrication. The main factors that influenced the values of band variances were identified. The performance of the distributors was found to correlate most strongly with the volume of the flow distributors. The separation bed should be positioned immediately after, but not against the flow distributor. It was concluded that BF distributors perform best in terms of band variance. The values of band variances for the BF distributor decreased with increasing angle between bifurcation branches and the lowest value of about 0.01 mm(2) was found for α=175°. Both BF and RI flow distributors were found to perform reasonably well when imperfections were present in the structure. However, severe blockages (exceeding 75% of the cross-sectional area and length) of channels in, especially, BF flow distributors may jeopardize their performance.

Multiple molecular forms of pyridinolines cross-links excreted in human urine evaluated by chromatographic and immunoassay methods.

The measurement of the collagen cross-links, hydroxylysylpyridinoline (HP) and lysylpyridinoline (LP), excreted in urine either in free or peptide-bound forms represents the most extensively investigated biochemical marker of bone collagen degradation. We studied the urinary molecular forms of pyridinolines after separation in free and peptide-linked fractions by chromatography and serial dialysis. The pyridinoline amounts of molecular species (free, < 1000 D, 1000-3500 D, 3500-10,000 D, and > 10,000 D) were evaluated by high performance liquid chromatography (HPLC) as well as with the two newly introduced enzyme-linked immunosorbent assay (ELISA) methods for determination of free pyridinolines (collagen Pyrilinks and collagen Pyrilinks-D). The variability of urinary pyridinoline forms were studied in healthy adult control subjects (n = 10, 38.4 +/- 7.5) years), in adolescents (n = 10, 16 +/- 3.3 years), and in elderly subjects with vitamin D insufficiency (n = 10, 87.3 +/- 4.3 years). Free and peptide-conjugated pyridinolines with MW < 1000 D constitute the major part of urinary cross-links in all groups, with a significantly lesser excretion in elderly patients than in adolescent groups. Expressed as a percent of total cross-links, urinary free pyridinolines assessed by direct HPLC are less in elderly subjects (HP = 34.2 +/- 6.2%, LP = 32.7 +/- 7.6%) than in adolescents (HP = 45.8 +/- 10.8%, p = 0.0065 and LP = 47.8 +/- 12.1%, p = 0.012) and in healthy adults (HP = 39.3 +/- 11.5%, NS and LP = 38.1 +/- 9.3%, NS).(ABSTRACT TRUNCATED AT 250 WORDS)

Markers of bone remodeling in the elderly subject: effects of vitamin D insufficiency and its correction.

The elderly subject is prone to both vitamin B insufficiency and calcium insufficiency due to a low calcium intake and calcium malabsorption. These two alterations may lead to secondary hyperparathyroidism, and thus to increased bone loss. We investigated 72 elderly subjects (16 men and 56 women) with vitamin D insufficiency and 25 healthy elderly women with normal vitamin D status, with respect to their indices of calcium metabolism and of bone remodeling: serum total alkaline phosphates (phosphatases), bone AP (BAP), osteocalcin (BGP), tartrate-resistant acid phosphatase (TRAP), urine hydroxyproline (HYP), and the 3-OH-pyridinium derivatives pyridinoline (PYD) and deoxypyridinoline (DPD), which are new markers of bone resorption. We then studied the modifications of these markers in the patients with vitamin D insufficiency at 3 months and 6 months after onset of a daily vitamin D and calcium supplementation. When compared with elderly subjects with normal vitamin D status, patients with vitamin D insufficiency had increased intact parathyroid hormone (iPTH) levels (60.1 +/- 10.2 vs 30.2 +/- 4.5, p < 0.001) and a high bone turnover as reflected by increased values of most serum and urine markers of bone remodeling. PYD and DPD levels were significantly correlated with all indices of bone turnover, unlike HYP, which showed no correlation with bone formation markers (AP, BAP, and BGP). A daily supplement of 800 IU vitamin D3 and 1 g of elemental calcium increased 25(OH)D levels and induced a dramatic decrease of iPTH levels; at 3 and 6 months, the mean iPTH level decreased by 50% (p < 0.0001), reaching the mean value of healthy vitamin D sufficient elderly women. All markers of bone turnover, except TRAP, decreased significantly at 3 and 6 months. The PYD/DPD ratio increased significantly at 3 and 6 months. The decrease of bone markers was more marked in patients with more severe hyperparathyroidism, the greatest variations being obtained with BAP (45%, p = 0.006) and DPD (43%, p = 0.036) levels. Most markers of bone remodeling are increased in elderly subjects with vitamin D insufficiently and vary with its correction. However, BAP and DPD are the most sensitive indicators of increased bone turnover due to secondary hyperparathyroidism.

Different responses of free and peptide-bound cross-links to vitamin D and calcium supplementation in elderly women with vitamin D insufficiency.

Recent findings have shown that bisphosphonates had different effects on the urinary excretion of free and peptide-bound cross-links. Because of this discrepancy, we investigated the effects of another antiresorptive therapy, i.e. vitamin D (vitD) and calcium (Ca) supplementation (800 IU vit D3 and 1 g elemental calcium daily for 6 months) in elderly women (n = 21, age: 83.5 +/- 1.5 yr) with vitD insufficiency and secondary hyperparathyroidism (mean level 25 hydroxy vitamin D = 3.17 +/- 1.2 ng/mL, mean level of intact parathormone = 45.3 +/- 22.7 pg/mL) on the urinary excretion of free and peptide-bound cross-links. A group of free-living, healthy elderly women (n = 25, age: 76.6 +/- 3.1 yr) with a normal vitD status (mean level of 25 OH D = 23.4 +/- 8.9 ng/mL, intact parathormone = 30.2 +/- 11.2 pg/mL) was simultaneously studied. Bone resorption was assessed by total (T), free (F), peptidyl (P) hydroxylysylpyridinoline (HP) and lysylpyridinoline (LP) measured with high performance liquid chromatography, by F-LP determined with enzyme linked immunosorbent assay (iF-LP) and by the N- and C-terminal telopeptides of type I collagen (NTX and Cross-laps) before and after (3 and 6 months) therapy. Comparison of the two groups of elderly women at baseline showed that the urinary excretion of pyridinoline cross-links (T, F, and peptide-bound forms) and of telopeptide fragment of type I collagen were all increased in patients with a low vitD status. Highly significant differences were seen principally for T-HP, F-HP, and F-LP (P < 0.001). Correlation studies between each marker showed that the values of pyridinoline cross-links (T and peptide-bound forms) and of the telopeptide fragments of type I collagen correlated well, but the correlation was slightly less pronounced between free pyridinolines and the other markers. After treatment, the response to therapy was greatest for peptide-bound cross-links assessed by high performance liquid chromatography and for telopeptide fragments of type I collagen (percent change at 6 months: -21% for P-HP P < 0.05, -26% for P-LP P < 0.05, -31% for NTX P < 0.01, and -51% for CLaps P < 0.001). In contrast, free pyridinolines excretion (F-HP and F-LP) assessed by high performance liquid chromatography as well as by immunoassay remained unchanged at 3 and 6 months. Because marked and significant changes were seen with peptide-bound cross-links only and not with free forms, we conclude that vitD and Ca therapy has the same effects as bisphosphonates on the urinary excretion of free and peptide-bound cross-links. So far, no rational mechanism can be given to explain this discrepancy, and further studies are needed before routine application of these bone collagen degradation products as bone resorption markers.

The possibility of generating high-speed shear-driven flows and their potential application in liquid chromatography

An experimental proof of principle is presented for the possibility to use a shear force field to generate a stable, chromatography enabling fluid flow through micrometer and submicrometer channels without the need for a pressure or a voltage gradient. In our setup, we were able to successfully move a color tracer plug at speeds exceeding 2 cm/s through a 0.125-microm-thick and 4-mm-wide channel, without creating a pressure drop or a pressure buildup. By showing that the speed of microchannel flows can be drastically increased by simply switching from one driving force to another, the presented experiments open the road to the development of a new type of chromatography, referred to as shear-driven chromatography, potentially offering unprecedented separation speeds and resolutions and complying perfectly with the present trend toward the miniaturization and parallelization of analytical separation equipment.

Evaluation of a new blood autotransfusion device.

A new autotransfusion device was evaluated in dogs. The device uses citrate phosphate dextrose as the blood anticoagulant and automatically delivers the agent in a ratio approximating that found in banked blood. Bleeding, aspiration, and autotransfusion of approximately 3 estimated blood volumes produced small changes in hematologic and coagulation studies. Blood electrolytes stayed within normal ranges. Activated clotting times stayed within normal range after autotransfusion of 2 blood volumes but increased slightly after 3 blood volume transfusions. No significant histopathologic changes were found in any organ system. Rapid infusion of citrated blood causes myocardial depression, which can be reversed by giving calcium. Overall performance of the device was excellent, suggesting further documentation in a clinical setting and evaluation with human blood.

[A simple method for the separation and purification of the surface antigen of hepatitis B from serum (author's transl)]. / Méthodologic simple de séparation et de purification de l'antigène de surface de l'hépatite B à partir du sérum

The present report describes a simple, rapid and reproducible procedure for preparation of hepatitis B surface antigen from human positive sera, with a satisfactory yield. After removal of beta- and alpha-lipoproteins by dextran sulphate precipitation and subsequent removal of IgM and a part of the other serum proteins by dialysis against distilled water, the supernatant obtained presents almost no change in its surface antigen content. After passing this solution through a molecular sieve of Biogel A-5 m, the surface antigen is detected in a well-defined peak. The fraction correspinding to this peak is collected in its totality. After lyophilization, it is passed through a molecular sieve of CPG 10, in phosphate buffer; this allows recovery of a substance which has retained its structural as well as its antigenic properties, and which is intact and practically free of any serum contamination.