European guideline (EuroGuiDerm) on atopic eczema - part II: non-systemic treatments and treatment recommendations for special AE patient populations.

The evidence- and consensus-based guideline on atopic eczema was developed in accordance with the EuroGuiDerm Guideline and Consensus Statement Development Manual. Four consensus conferences were held between December 2020 and July 2021. Twenty-nine experts (including clinicians and patient representatives) from 12 European countries participated. This second part of the guideline includes recommendations and detailed information on basic therapy with emollients and moisturizers, topical anti-inflammatory treatment, antimicrobial and antipruritic treatment and UV phototherapy. Furthermore, this part of the guideline covers techniques for avoiding provocation factors, as well as dietary interventions, immunotherapy, complementary medicine and educational interventions for patients with atopic eczema and deals with occupational and psychodermatological aspects of the disease. It also contains guidance on treatment for paediatric and adolescent patients and pregnant or breastfeeding women, as well as considerations for patients who want to have a child. A chapter on the patient perspective is also provided. The first part of the guideline, published separately, contains recommendations and guidance on systemic treatment with conventional immunosuppressive drugs, biologics and janus kinase (JAK) inhibitors, as well as information on the scope and purpose of the guideline, and a section on guideline methodology.

European guideline (EuroGuiDerm) on atopic eczema: part I - systemic therapy.

The evidence- and consensus-based guideline on atopic eczema was developed in accordance with the EuroGuiDerm Guideline and Consensus Statement Development Manual. Four consensus conferences were held between December 2020 and July 2021. Twenty-nine experts (including clinicians and patient representatives) from 12 European countries participated. This first part of the guideline includes general information on its scope and purpose, the health questions covered, target users and a methods section. It also provides guidance on which patients should be treated with systemic therapies, as well as recommendations and detailed information on each systemic drug. The systemic treatment options discussed in the guideline comprise conventional immunosuppressive drugs (azathioprine, ciclosporin, glucocorticosteroids, methotrexate and mycophenolate mofetil), biologics (dupilumab, lebrikizumab, nemolizumab, omalizumab and tralokinumab) and janus kinase inhibitors (abrocitinib, baricitinib and upadacitinib). Part two of the guideline will address avoidance of provocation factors, dietary interventions, immunotherapy, complementary medicine, educational interventions, occupational and psychodermatological aspects, patient perspective and considerations for paediatric, adolescent, pregnant and breastfeeding patients.

Effect of shoeing conditions on hoof dimensions in Icelandic and Warmblood horses.

In Icelandic horses (ICE), high, long hooves with added weights are often used to improve gait quality in competitions. From a biomechanical point of view, excessively long hooves can be problematic as they increase forces acting on the distal limb. The study aimed to determine if hoof capsule size and angle of ICE at competitions are comparable to those of Warmblood horses (WB) requiring shoeing. Hoof dimensions and balance were quantified in 13 horses of each breed at the beginning of a shoeing period (BSP), allowing a baseline comparison between breeds, as well as for WB at the end of a shoeing period (ESP) and for ICE when shod for competition (COMP). Left front and hind hoof capsules and bony structures of the hooves were measured on lateromedial and dorsopalmar/dorsoplantar radiographs at BSP (ICE, WB), ESP (WB) and COMP (ICE), respectively, using the software Metron-Hoof-Pro. Compared to WBBSP, ICEBSP had larger hoof capsules relative to the size of P3 and, when controlled for withers height, longer dorsal hoof wall lengths. The comparison of ICECOMP to WBESP indicated that the relative increase in hoof length and volume was greater in ICECOMP compared to WBESP. Hoof balance was closer to previously published values in WBs, while hoof angles in ICE (particularly palmar/plantar angle of P3) were generally larger and thus less problematic. In conclusion, hooves of ICECOMP were relatively larger and less balanced than WBESP, but hoof angles in WB were smaller than in ICE.

Hoof conformation in Icelandic competition horses and its interrelationship with hoof pathologies and tölt performance.

In order to improve the four-beat-rhythm and forelimb action of the tölt in Icelandic horses (ICE), special shoeing techniques are applied particularly in the front hooves by increasing hoof length and height, or by adding weight to the distal limbs. Although regulations limit dorsal hoof wall length (LDHW) in competition ICE, their shoeing often deviates from a biomechanically optimal distal limb conformation. This study aimed to qualitatively and quantitatively describe current shoeing practices of ICE in competition. Moreover, the influence of LDHW on the occurrence of deviations from a standard hoof conformation, as well as on tölt performance was investigated. At four European competition sites, hoof dimensions of 133 randomly selected ICE were measured manually, and limb conformation and hoof balance were described. Dorsopalmar/-plantar and lateromedial radiographs of the left front and hind hooves were taken of each horse. Various parameters related to hoof dimensions and balance were measured on radiographs using the Metron-Hoof-Pro software. Flares of the dorsal hoof wall had the highest prevalence of all investigated hoof deformities. Multiple logistic regression analysis revealed that longer LDHW was associated with the occurrence of toe flares (P = 0.045), a broken hoof pastern axis (P = 0.003), and asymmetry of the quarter wall heights (P = 0.015). However, horses with a longer LDHW achieved higher scores in competition. In spite of its positive effect on tölt performance, a long LDHW is not recommendable as it may be associated with a higher prevalence of certain hoof deformities.

Review: Comparative methane production in mammalian herbivores.

Methane (CH4) production is a ubiquitous, apparently unavoidable side effect of fermentative fibre digestion by symbiotic microbiota in mammalian herbivores. Here, a data compilation is presented of in vivo CH4 measurements in individuals of 37 mammalian herbivore species fed forage-only diets, from the literature and from hitherto unpublished measurements. In contrast to previous claims, absolute CH4 emissions scaled linearly to DM intake, and CH4 yields (per DM or gross energy intake) did not vary significantly with body mass. CH4 physiology hence cannot be construed to represent an intrinsic ruminant or herbivore body size limitation. The dataset does not support traditional dichotomies of CH4 emission intensity between ruminants and nonruminants, or between foregut and hindgut fermenters. Several rodent hindgut fermenters and nonruminant foregut fermenters emit CH4 of a magnitude as high as ruminants of similar size, intake level, digesta retention or gut capacity. By contrast, equids, macropods (kangaroos) and rabbits produce few CH4 and have low CH4 : CO2 ratios for their size, intake level, digesta retention or gut capacity, ruling out these factors as explanation for interspecific variation. These findings lead to the conclusion that still unidentified host-specific factors other than digesta retention characteristics, or the presence of rumination or a foregut, influence CH4 production. Measurements of CH4 yield per digested fibre indicate that the amount of CH4 produced during fibre digestion varies not only across but also within species, possibly pointing towards variation in microbiota functionality. Recent findings on the genetic control of microbiome composition, including methanogens, raise the question about the benefits methanogens provide for many (but apparently not to the same extent for all) species, which possibly prevented the evolution of the hosting of low-methanogenic microbiota across mammals.

Effects of replacing maize silage with lucerne silage and lucerne silage chop length on rumen function and milk fatty acid composition.

The objective of this study was to investigate whether higher lucerne (Medicago sativa; alfalfa) silage inclusion rate and longer lucerne chop length improves rumen function through increased provision of physically effective fiber, when included in a maize and lucerne silage-based total mixed ration. Diets were formulated to contain a 50:50 forage:concentrate ratio [dry matter (DM) basis] and be isonitrogenous and contain equal levels of neutral detergent fiber (320 g/kg). The forage portion of the offered diets was composed of maize and lucerne silage DM in proportions (wt/wt) of either 25:75 (high lucerne; HL) or 75:25 (low lucerne; LL). Second-cut lucerne was harvested and conserved as silage at either a long (L) or a short (S) chop length (geometric mean particle lengths of 9.0 and 14.3 mm, respectively). These variables were combined in a 2 × 2 factorial arrangement to give 4 treatments (HLL, HLS, LLL, LLS), which were fed in a 4 × 4 Latin square design study to 4 rumen-cannulated, multiparous, Holstein dairy cows in mid lactation. Effects on DM intake, chewing behavior, rumen volatile fatty acid concentration, rumen pH, rumen and fecal particle size, milk production, and milk fatty acid profile were measured. Longer chop length increased rumination times per kilogram of DM intake (+2.8 min/kg) relative to the S chop length, with HLL diets resulting in the most rumination chews. Rumen concentrations of total volatile fatty acids, acetate, and n-valerate were higher for the HLS diet than the other 3 diets, whereas rumen propionate concentration was lowest for the HLL diet. Physically effective fiber (particles >4 mm) percentage in the rumen mat was increased when L chop length was fed regardless of lucerne inclusion rate. No effect of treatment was observed for milk yield, although milk protein concentration was increased by L for the LL diet (+1.6 g/kg) and decreased by L for the HLL diet (-1.4 g/kg). Milk fat concentrations of total cis-18:1 (+3.7 g/100 g of fatty acids) and 18:3 n-3 (+0.2 g/100 g of fatty acids) were greater with HL. In conclusion, longer lucerne silage chop length increased time spent ruminating per kilogram of DM intake, but had no effect on rumen pH in the present study. Increasing dietary lucerne inclusion rate had no effects on rumination activity or rumen pH, but decreased the ratio of n-6:n-3 polyunsaturated fatty acid concentrations in milk fat.

Retention of solute and particle markers in the digestive tract of chinchillas (Chinchilla laniger).

The chinchilla (Chinchilla laniger) is a herbivorous hystricomorph South American rodent for which no mean digesta retention times have been reported so far. Six animals (mean body mass ± standard deviation: 513 ± 99 g) on a diet of grass hay and lucerne-based pellets were given a pulse dose of a solute (cobalt-EDTA) and a particle (chromium-mordanted fibre, <2 mm) marker with subsequent frequent faecal collection. Dry matter intake was 45.2 ± 8.0 g/kg(0.75) /day. Mean retention times were 22.2 ± 5.3 h for solutes and 25.4 ± 5.2 h for particles, with the difference being not significant within individuals. This indicates the presence of a 'mucus-trap' colonic separation mechanism, which is in accord with morphological descriptions of the typical colonic furrow in chinchillas. Corresponding to a strategy of colonic digesta separation and caecotroph formation, secondary marker excretion peaks indicated coprophagic events that were spaced approximately 12 h apart. Given that these retention times appear longer than measures reported for rabbits (Oryctolagus cuniculus) or guinea pigs (Cavia procellus), it would be interesting to compare the digestive efficiency of chinchillas on high levels of dietary fibre to other species.

Digestive physiology, metabolism and methane production of captive Linné's two-toed sloths (Choloepus didactylus).

Sloths are renowned for their low metabolic rate, low food intake and low defecation frequency. We investigated factors of digestive physiology and energy metabolism in four captive individuals (mean body mass 10.0 ± SD 3.7 kg) of a hitherto mostly unstudied sloth species, Linné's two-toed sloth (Choloepus didactylus), in a 2-week digestion recording and 23-h respiration experiment on animals fed a standard zoo diet of vegetables and starchy components. Dry matter intake, defecation frequency and particle mean retention time (MRT) in the gastrointestinal tract (GIT) were 12 ± 3 g/(kg(0.75)  day), once every 5 days and >140 h in three individuals, but 53 g/(kg(0.75)  day), daily and 82 h in one individual that was apparently compensating for a period of weight loss prior to the experiment. In all animals, solute marker was eliminated at a faster rate than the particle marker, indicating 'digesta washing' in the sloths' GIT. The overall metabolic rate calculated from oxygen consumption matched the metabolisable energy intake in three individuals [173 ± 22 vs. 168 ± 44 kJ/(kg(0.75)  day)] but not in the fourth one [225 vs. 698 kJ/(kg(0.75)  day)], supporting the interpretation that this animal was replenishing body stores. In spite of the low food intake and the low-fibre diet (209 ± 26 g neutral detergent fibre/kg dry matter), methane production was rather high accounting for 9.4 ± 0.8% of gross energy intake (2.7% in the fourth individual), which exceeded literature data for ruminants on forage-only diets. These results corroborate literature reports on low intake, low defecation frequency, low metabolic rate and long MRT in other sloth species. The long MRT is probably responsible for the comparatively high methane production, providing more opportunity for methanogenic archaea than in other non-ruminant mammals to produce significant amounts of methane.

Quantification aspects of constant pressure (ultra) high pressure liquid chromatography using mass-sensitive detectors with a nebulizing interface.

The present contribution investigates the quantitation aspects of mass-sensitive detectors with nebulizing interface (ESI-MSD, ELSD, CAD) in the constant pressure gradient elution mode. In this operation mode, the pressure is controlled and maintained at a set value and the liquid flow rate will vary according to the inverse mobile phase viscosity. As the pressure is continuously kept at the allowable maximum during the entire gradient run, the average liquid flow rate is higher compared to that in the conventional constant flow rate operation mode, thus shortening the analysis time. The following three mass-sensitive detectors were investigated: mass spectrometry detector (MS), evaporative light scattering detector (ELSD) and charged aerosol detector (CAD) and a wide variety of samples (phenones, polyaromatic hydrocarbons, wine, cocoa butter) has been considered. It was found that the nebulizing efficiency of the LC-interfaces of the three detectors under consideration changes with the increasing liquid flow rate. For the MS, the increasing flow rate leads to a lower peak area whereas for the ELSD the peak area increases compared to the constant flow rate mode. The peak area obtained with a CAD is rather insensitive to the liquid flow rate. The reproducibility of the peak area remains similar in both modes, although variation in system permeability compromises the 'long-term' reproducibility. This problem can however be overcome by running a flow rate program with an optimized flow rate and composition profile obtained from the constant pressure mode. In this case, the quantification remains reproducibile, despite any occuring variations of the system permeability. Furthermore, the same fragmentation pattern (MS) has been found in the constant pressure mode compared to the customary constant flow rate mode.

Comparison of the quantitative performance of constant pressure versus constant flow rate gradient elution separations using concentration-sensitive detectors.

This contribution discusses the difference in chromatographic performance when switching from the customary employed constant flow rate gradient elution mode to the recently re-introduced constant pressure gradient elution mode. In this mode, the inlet pressure is maintained at a set value even when the mobile phase viscosity becomes lower than the maximum mobile phase viscosity encountered during the gradient program. This leads to a higher average flow rate compared to the constant flow rate mode and results in a shorter analysis time. When both modes carry out the same mobile phase gradient program in volumetric units, normally identical selectivities are obtained. However, small deviations in selectivity are found due to the differences in pressure and viscous heating effects. These selectivity differences are of the same type as those observed when switching from HPLC to UHPLC and are inevitable when speeding up the analysis by applying a higher pressure. It was also found that, when using concentration-sensitive detectors, the constant pressure elution mode leads to identical peak areas as the constant flow rate mode. Also the linearity is maintained. In addition, the repeatability of the peak area and retention time remains the same when switching between both elution modes.

Kinetic performance limits of constant pressure versus constant flow rate gradient elution separations. Part I: theory.

We report on a general theoretical assessment of the potential kinetic advantages of running LC gradient elution separations in the constant-pressure mode instead of in the customarily used constant-flow rate mode. Analytical calculations as well as numerical simulation results are presented. It is shown that, provided both modes are run with the same volume-based gradient program, the constant-pressure mode can potentially offer an identical separation selectivity (except from some small differences induced by the difference in pressure and viscous heating trajectory), but in a significantly shorter time. For a gradient running between 5 and 95% of organic modifier, the decrease in analysis time can be expected to be of the order of some 20% for both water-methanol and water-acetonitrile gradients, and only weakly depending on the value of V(G)/V0 (or equivalently t(G)/t0). Obviously, the gain will be smaller when the start and end composition lie closer to the viscosity maximum of the considered water-organic modifier system. The assumptions underlying the obtained results (no effects of pressure and temperature on the viscosity or retention coefficient) are critically reviewed, and can be inferred to only have a small effect on the general conclusions. It is also shown that, under the adopted assumptions, the kinetic plot theory also holds for operations where the flow rate varies with the time, as is the case for constant-pressure operation. Comparing both operation modes in a kinetic plot representing the maximal peak capacity versus time, it is theoretically predicted here that both modes can be expected to perform equally well in the fully C-term dominated regime (where H varies linearly with the flow rate), while the constant pressure mode is advantageous for all lower flow rates. Near the optimal flow rate, and for linear gradients running from 5 to 95% organic modifier, time gains of the order of some 20% can be expected (or 25-30% when accounting for the fact that the constant pressure mode can be run without having to leave a pressure safety margin of 5-10% as is needed in the constant flow rate mode).

Kinetic performance limits of constant pressure versus constant flow rate gradient elution separations. Part II: experimental.

We report on a first series of experiments comparing the selectivity and the kinetic performance of constant flow rate and constant pressure mode gradient elution separations. Both water-methanol and water-acetonitrile mobile phase mixtures have been considered, as well as different samples and gradient programs. Instrument pressures up to 1200 bar have been used. Neglecting some small possible deviations caused by viscous heating effects, the experiments could confirm the theoretical expectation that both operation modes should lead to identical separation selectivities provided the same mobile phase gradient program is run in reduced volumetric coordinates. Also in agreement with the theoretical expectations, the cP-mode led to a gain in analysis time amounting up to some 17% for linear gradients running from 5 to 95% of organic modifier at ultra-high pressures. Gains of over 25% were obtained for segmented gradients, at least when the flat portions of the gradient program were situated in regions where the gradient composition was the least viscous. Detailed plate height measurements showed that the single difference between the constant flow rate and the constant pressure mode is a (small) difference in efficiency caused by the difference in average flow rate, in turn leading to a different intrinsic band broadening. Separating a phenone sample with a 20-95% water-acetonitrile gradient, the cP-mode leads to gradient plate heights that are some 20-40% smaller than in the cF-mode in the B-term dominated regime, while they are some 5-10% larger in the C-term dominated regime. Considering a separation with sub 2-µm particles on a 350 mm long coupled column, switching to the constant pressure mode allowed to finish the run in 29 instead of in 35 min, while also a larger peak capacity is obtained (going from 334 in the cF-mode to 339 in the cP-mode) and the mutual selectivity between the different peaks is fully retained.

Towards a solution for viscous heating in ultra-high pressure liquid chromatography using intermediate cooling.

A generic solution is proposed for the deleterious viscous heating effects in adiabatic or near-adiabatic systems that can be expected when trying to push the column operating pressures above the currently available range of ultra-high pressures (i.e., 1200 bar). A set of proof-of-principle experiments, mainly using existing commercial equipment, is presented. The solution is based on splitting up a column with given length L into n segments with length L/n, and providing an active cooling to the capillaries connecting the segments. In this way, the viscous heat is removed at a location where the radial heat removal does not lead to an efficiency loss (i.e., in the thin connection capillaries), while the column segments can be operated under near-adiabatic conditions without suffering from an unacceptable rise of the mobile phase temperature. Experimental results indicate that the column segmentation does not lead to a significant efficiency loss (comparing the performance of a 10 cm column with a 2 cm x 5 cm column system), whereas, as expected, the system displays a much improved temperature stability, both in time (because of the shortened temperature transient times) and in space (reduction of the average axial temperature rise by a factor n). The method also prevents a large backflow of heat along the column wall that would lead to large efficiency losses if one would attempt to operate columns at pressures of 1500 bar or more. A real-world pharmaceutical example is given where this improved temperature robustness could help in moderating the changes in selectivity during method transfer from a low to a high pressure operation, although the complex non-linear behavior of the viscous heating and high pressure effects result in lower than expected improvement.

Separation of basic solutes by reversed-phase capillary electrochromatography.

The separation of basic solutes at low pH by capillary electrochromatography (CEC) has been investigated. The feasibility of separation of basic solutes by CEC was demonstrated. Influence of operational parameters, solvent composition, pH, temperature on retention and selectivity of the separation of a mixture of basic, neutral and acidic drug standards has been investigated. The observed elution behavior has been modeled to account for both chromatographic retention and differential electrophoretic mobility of the solutes. This model was verified experimentally. It is demonstrated in this work that the elution window of solutes in reversed-phase CEC is expanded to range from -1 to infinity.

Towards the column bed stabilization of columns in capillary electroendosmotic chromatography. Immobilization of microparticulate silica columns to a continuous bed.

This article discusses a novel method generating a continuous bed inside the CEC column. The column bed composed of microparticulate reversed-phase silica is completely immobilized by a hydrothermal treatment using water for the immobilization process. This process eliminates the manufacture of frits of both ends of the column and all problems associated with their preparation. Fundamental studies on operational parameters will be presented such as the dependence of the immobilization on the column temperature, the type of stationary phase and the column back pressure. The immobilized CEC columns show the same high column efficiency as packed columns with frits.

A microfluidic system for high-speed reproducible DNA sizing and quantitation.

Microfabrication technology was used to develop a system consisting of disposable glass chips containing etched channels, reagents including polymer matrix and size standards, computer-controlled instrumentation for performing electrophoretic separations and fluorescence detection of double-stranded DNA, and software for automated data analysis. System performance was validated for separation and quantitation reproducibility using samples varying in amount and size of DNA fragments, buffer composition, and salt concentrations. Several applications of the microfluidic system for DNA analysis have been demonstrated, such as of polymerase chain reaction (PCR) products, sizing of plasmid digests, and detection of point mutations by restriction fragment length polymorphism (RFLP) mapping.

[New results in the visualization of the spinal dura mater with scanning electron microscopy]. / Neue Ergebnisse bei der Darstellung der Dura mater spinalis mittels Rasterelektronenmikroskopie.

UNLABELLED: Although there are various published descriptions of the dura mater spinalis [4, 7, 9, 11, 16], some points relating to the texture of the collagen fibres in the dura have still not been adequately explained. In this study the orientation of the collagen fibril bundles was revealed with the aid of scanning electron microscopy, and our observations have yielded new insights into the three-dimensional structure of the human dura mater spinalis. MATERIALS AND METHODS: The preparations used were taken from the bodies of four persons who had died of acute cardiac infarct at the ages of 70-78 years. The histories of these patients gave no indications of earlier neurological, endocrine or septic illnesses. The tissue examined was taken 8-12 h after death in all cases; it was immediately fixed in glutaraldehyde and then processed for scanning electron microscopy in the usual way. RESULTS: In the outermost (epidural) layer of the dura mater spinalis the collagen fibres are bunched together in bands that run in all directions. Elastic fibres 2 mm thick are woven into this three-dimensional network of collagen systems. On the inside (the arachnoid side) thin collagen fibres are fused into layers in such a way that the innermost layer resting on the arachnoid has a smooth, shiny appearance comparable to that of a serosa. It is attached to the actual dura with a supporting band of connective tissue. Rests of the subdural neuroepithelium could contribute to the smooth appearance of the superficial aspect. CONCLUSIONS: The outermost layer of the dura is made up mainly of collagen fibres, which run in all three directions--longitudinal, horizontal and transverse--both singly and in groups. These findings are at odds with "classic" descriptions, according to which the fibres in the dura mater spinalis all have a parallel course with a longitudinal orientation in tangential sections.

New perspectives in the microscopic structure of human dura mater in the dorsolumbar region.

BACKGROUND AND OBJECTIVES: The object of this study was to describe the three-dimensional structure of the dura mater by use of scanning electron microscopy. METHODS: Microscopic dissection of the dura mater from four fresh cadavers (aged 70, 75, 76, and 80 years) 8-12 hours after death were investigated in three different planes (longitudinal, tangential, and transverse). RESULTS: The external surface of the dura mater, facing the epidural space, consisted of a network of randomly oriented fine collagen fibers. The thicker elastic fibers (2 microns in diameter) were observed on the surface of the dura. In the inner part of the dura mater, there were very fine lamellae of collagen fibers, which were bundled into thicker (4-5 microns) layers. The dura mater consisted of 78-82 layers, each layer including 8-12 very fine lamellae. CONCLUSIONS: The fibers of the dura mater do not run in a longitudinal direction and are not arranged in a parallel fashion. Cytoarchitecturally the dura mater is a laminated structure built up from well-defined layers oriented concentrically around the medulla spinalis.