Binding and inactivation of human coronaviruses, including SARS-CoV-2, onto purified clinoptilolite-tuff.

The current COVID19 pandemic is caused by a positive-sense single-stranded RNA virus, which presents high mutational rates. The development of effective therapeutics and mitigation strategies using vaccination or therapeutic antibodies faces serious challenges because of the regular emergence of immune escape variants of the virus. An efficient approach would involve the use of an agent to non-specifically limit or block viruses contacting the mucosae and therefore entering the body. Here, we investigated the ability of a micronized purified clinoptilolite-tuff to bind and neutralize different viruses from the Coronaviridae family. Using plaque assay, RT-qPCR and immunostaining, the adsorption and inactivation of the seasonal human coronavirus HCoV-229E and of 2 SARS-CoV-2 variants were demonstrated. The resulting data suggest that purified clinoptilolite-tuff could be used as an ingredient in new medical devices and/or pharmaceuticals to prevent or mitigate SARS-CoV-2 dissemination.

Evaluation of a microbial muramidase supplementation on growth performance, apparent ileal digestibility, and intestinal histology of broiler chickens.

The current study evaluated the effects of different inclusion levels of microbial muramidase (Muramidase 007, DSM Nutritional Products) on gastrointestinal functionality by determination of apparent ileal digestibility (AID) of nutrients, investigation of intestinal histomorphology, and quantification of resulting growth performance. Four maize-wheat-soybean experimental diets were produced without (C) and with different dosages of muramidase: low (L, 25,000 LSU(F)/kg), medium (M, 35,000 LSU(F)/kg), and high (H, 45,000 LSU(F)/kg); diets were fed to broilers for 35 d. At the end of the experiment, AID of ether extract (EE), crude protein (CP), Ca, and P were determined and samples of the mid-jejunum and -ileum were collected for histomorphological observations. Data were subjected to ANOVA analysis using the GLM procedure. Orthogonal polynomial contrasts were used to assess linear and quadratic effects of different levels of the muramidase. At the end of the trial, Muramidase 007 supplementation linearly increased body weight gain and decreased feed conversion ratio (FCR) (P ≤ 0.05). Adding the muramidase to broiler diets also linearly increased the European poultry efficiency factor (P ≤ 0.05). Inclusion of the muramidase in broiler diets linearly increased AID of CP, EE, and P (P ≤ 0.05), and the H group had a higher AID of EE and CP compared to C group (P ≤ 0.05). Microbial muramidase supplementation linearly increased ileal villus length to crypt depth ratio and decreased the number of ileal CD45 cells (P ≤ 0.05). Broilers fed M and H diets had fewer number of CD45 cells in the ileum compared to those in C group (P ≤ 0.05). In conclusion, the results of the present study demonstrated that inclusion of the microbial muramidase in broiler diets could increase AID of key nutrients and improve growth performance in broilers. Adding microbial muramidase to broiler diets can therefore be considered as an interesting prospect to improve gastrointestinal functionality. Biological mechanisms causing these improvements need to be studied further.

High-energy x-ray Talbot-Lau radiography of a human knee.

We report on a radiographic measurement of an ex vivo human knee using a grating-based phase-contrast imaging setup and a medical x-ray tube at a tube voltage of 70 kV. The measurement has been carried out using a Talbot-Lau setup that is suitable to achieve a high visibility in the energy regime of medical imaging. In a medical reading by an experienced trauma surgeon signatures of chondrocalcinosis in the medial meniscus have been identified more evidently using the dark-field image in comparison to the conventional attenuation image. The analysis has been carried out at various dose levels down to 0.14 mGy measured as air kerma, which is a dose comparable to clinically used radiographic devices. The diagnosis has been confirmed by a histological analysis of the meniscus tissue. In the introduced high-frequency filtered phase-contrast image the anterior and posterior horn of the medial meniscus and the posterior cruciate ligament have also been visible. Furthermore, atherosclerotic plaque is visible in both imaging modalities, attenuation and dark-field, despite the presence of overlaying bone. This measurement, for the first time, proves the feasibility of Talbot-Lau x-ray imaging at high-energy spectra above 40 kVp and reasonable dose levels with regard to spacious and dense objects.

Femoral rotation in total knee arthroplasty: a comparison of patient individualized jigs with gap balancing in relation to anatomic landmarks.

PURPOSE: The purpose of our study was to compare the accuracy of the rotational position of the femoral component in total knee arthroplasty aligned with patient individualized jigs (PSJ) to a gap balancing technique (GBT). METHODS: A consecutive series of 21 osteoarthritic patients were treated with 22 cruciate-retaining total knee prostheses. During surgery, the rotation of the femoral component pinholes was recorded for all knees using PSJ and GBT and transferred to computer tomograms (CT). The rotational differences between PSJ and GBT relative to the transepicondylar axis were analysed. RESULTS: The medium rotation of the femoral component pinholes was 1.3° ± 5.1° (min = -6.3°; max = 14.4°) for PSJ and 0.1 ± 1.4° (min = -1.6°; max = 3.4°) for GBT. Outliers of more than 3° were found more frequently with PSJ in 12 cases but only in one for GBT. CONCLUSION: Based on our study, we would not recommend relying intra-operatively solely on the CT-based PSJ without the option to adjust or control femoral rotation. LEVEL OF EVIDENCE: II.

Optimisation of image reconstruction for phase-contrast x-ray Talbot-Lau imaging with regard to mechanical robustness.

X-ray grating-based phase-contrast imaging opens new opportunities, inter alia, in medical imaging and non-destructive testing. Because, information about the attenuation properties and about the refractive properties of an object are gained simultaneously. Talbot-Lau imaging requires the knowledge of a reference or free-field image. The long-term stability of a Talbot-Lau interferometer is related to the time span of the validity of a measured reference image. It would be desirable to keep the validity of the reference image for a day or longer to improve feasibility of Talbot-Lau imaging. However, for example thermal and other long-term external influences result in drifting effects of the phase images. Therefore, phases are shifting over time and the reference image is not valid for long-term measurements. Thus, artifacts occur in differential phase-contrast images. We developed an algorithm to determine the differential phase-contrast image with the help of just one calibration image, which is valid for a long time-period. With the help of this algorithm, called phase-plane-fit method, it is possible to save measurement-time, as it is not necessary to take a reference image for each measurement. Additionally, transferring the interferometer technique from laboratory setups to conventional imaging systems the necessary rigidity of the system is difficult to achieve. Therefore, short-term effects like vibrations or distortions of the system lead to imperfections within the phase-stepping procedure. Consequently, artifacts occur in all three image modalities (differential phase-contrast image, attenuation image and dark-field image) of Talbot-Lau imaging. This is a problem with regard to the intended use of phase-contrast imaging for example in clinical routine or non-destructive testing. In this publication an algorithm of Vargas et al is applied and complemented to correct inaccurate phase-step positions with the help of a principal component analysis (PCA). Thus, it is possible to calculate the artifact free images. Subsequently, the whole algorithm is called PCA minimization algorithm.

Increasing the field of view in grating based X-ray phase contrast imaging using stitched gratings.

Grating based X-ray differential phase contrast imaging (DPCI) allows for high contrast imaging of materials with similar absorption characteristics. In the last years' publications, small animals or parts of the human body like breast, hand, joints or blood vessels have been studied. Larger objects could not be investigated due to the restricted field of view limited by the available grating area. In this paper, we report on a new stitching method to increase the grating area significantly: individual gratings are merged on a carrier substrate. Whereas the grating fabrication process is based on the LIGA technology (X-ray lithography and electroplating) different cutting and joining methods have been evaluated. First imaging results using a 2×2 stitched analyzer grating in a Talbot-Lau interferometer have been generated using a conventional polychromatic X-ray source. The image quality and analysis confirm the high potential of the stitching method to increase the field of view considerably.

Guía oficial de la Sociedad Española de Neurología de práctica clínica en epilepsia / The Spanish Neurological Society official clinical practice guidelines in epilepsy

Las anteriores Guías oficiales de práctica clínica en epilepsia elaboradas por el Grupo de Estudio de Epilepsia de la Sociedad Española de Neurología (GE-SEN) estaban basadas en la opinión de expertos. La actual Guía de práctica clínica (GPC) en epilepsia se basa en el método científico que extrae recomendaciones a partir de evidencias científicas constatadas. Su principal función es disminuir la variabilidad de la práctica clínica a través de la homogeneización de la práctica médica. Alcance y objetivos: Esta GPC se centra en la atención integral de personas afectadas por una epilepsia, como síntoma principal y predominante, independiente de la edad de inicio y ámbito asistencial. Metodología: 1) Constitución del grupo de trabajo integrado por neurólogos del GE-SEN, con la colaboración de neuropediatras, neurofisiólogos y neurorradiólogos; 2) determinación de los aspectos clínicos a cubrir: diagnóstico, pronóstico y tratamiento; 3) búsqueda y selección de la evidencia científica relevante; 4) formulación de recomendaciones basadas en la clasificación de las evidencias científicas disponibles. Resultados: Contienen 192 recomendaciones. El 57% son de consenso entre autores y editores, como consecuencia del desconocimiento en muchos campos de esta patología. Conclusiones: Esta GPC, en epilepsia, con una metodología formal y rigurosa en la búsqueda de evidencias explícitas donde ha sido posible, formula recomendaciones extraídas de las mismas. En este artículo incluimos el capítulo de la GPC dedicado a situaciones de urgencia en crisis epilépticas y epilepsia, que pueden presentarse como una primera crisis epiléptica, una evolución desfavorable en un paciente con una epilepsia conocida o en su forma más grave como un estado epiléptico

Previous Official Clinical Practice Guidelines (CPGs) in Epilepsy were based on expert opinions and developed by the Epilepsy Study Group of the Spanish Neurological Society (GE-SEN). The current CPG in epilepsy is based on the scientific method, which extracts recommendations from published scientific evidence. A reduction in the variability in clinical practice through standardization of medical practice has become its main function. Scope and objectives: This CPG is focused on comprehensive care for individuals affected by epilepsy as a primary and predominant symptom, regardless of the age of onset and medical policy. Methodology: 1. Creation of GE-SEN neurologists working group, in collaboration with Neuropediatricians, Neurophysiologists and Neuroradiologists. 2. Identification of clinical areas to be covered: diagnosis, prognosis and treatment. 3. Search and selection of the relevant scientific evidence. 4. Formulation of recommendations based on the classification of the available scientific evidence. Results: It contains 161 recommendations of which 57% are consensus between authors and publishers, due to an important lack of awareness in many fields of this pathology. Conclusions: This Epilepsy CPG formulates recommendations based on explicit scientific evidence as a result of a formal and rigorous methodology, according to the current knowledge in the pre-selected areas. This paper includes the CPG chapter dedicated to emergency situations in seizures and epilepsy, which may present as a first seizure, an unfavorable outcome in a patient with known epilepsy, or status epilepticus as the most severe manifestation

[Switch of methoxy-polyethylene-glycol-epoetin beta to darbepoetin alfa in 263 dialysis patients]. / Étude de la conversion du méthoxypolyéthylèneglycol-époétine bêta vers la darbépoétine alfa chez 263 patients hémodialysés.

In early 2012, due to national supply disruption, the methoxy-polyethylene glycol-epoetin beta (CERA) was no longer available and has been replaced by darbepoetin alfa (DA) in all dialysis patients. Official recommendations for the replacement of one by the other is missing or unclear. On this occasion, we wanted to examine how the shift from CERA to DA was done in terms of dose conversion factor and the other factors that could have influenced the dose of DA prescribed (hemoglobin, patient weight, dose of CERA). This retrospective multicenter open conducted in six dialysis centers in Alsace is the first large study (n=263) that evaluated the switch from CERA to DA in all chronic hemodialysis patients. We found that the instantaneous ratio of dose adjustment is close to 1 and that nephrologists are mainly based on the dose of CERA for determining the DA dose, before hemoglobin and weight. However, establishing a true dose-response ratio between the two molecules requires a long term prospective study.

The Spanish Neurological Society official clinical practice guidelines in epilepsy.

Previous Official Clinical Practice Guidelines (CPGs) in Epilepsy were based on expert opinions and developed by the Epilepsy Study Group of the Spanish Neurological Society (GE-SEN). The current CPG in epilepsy is based on the scientific method, which extracts recommendations from published scientific evidence. A reduction in the variability in clinical practice through standardization of medical practice has become its main function. SCOPE AND OBJECTIVES: This CPG is focused on comprehensive care for individuals affected by epilepsy as a primary and predominant symptom, regardless of the age of onset and medical policy. METHODOLOGY: 1. Creation of GE-SEN neurologists working group, in collaboration with Neuropediatricians, Neurophysiologists and Neuroradiologists. 2. Identification of clinical areas to be covered: diagnosis, prognosis and treatment. 3. Search and selection of the relevant scientific evidence. 4. Formulation of recommendations based on the classification of the available scientific evidence. RESULTS: It contains 161 recommendations of which 57% are consensus between authors and publishers, due to an important lack of awareness in many fields of this pathology. CONCLUSIONS: This Epilepsy CPG formulates recommendations based on explicit scientific evidence as a result of a formal and rigorous methodology, according to the current knowledge in the pre-selected areas. This paper includes the CPG chapter dedicated to emergency situations in seizures and epilepsy, which may present as a first seizure, an unfavorable outcome in a patient with known epilepsy, or status epilepticus as the most severe manifestation.

Effects of high levels of dietary zinc oxide on ex vivo epithelial histamine response and investigations on histamine receptor action in the proximal colon of weaned piglets.

The aim of the study was to identify the effect of high dietary zinc oxide (ZnO) levels on the histamine-induced secretory-type response and histamine metabolism in the porcine proximal colon. After weaning at d 26, 3 diets with low (LZn), normal (NZn), and high (HZn) concentrations of zinc (57, 164, or 2,425 mg/kg) were fed to a total of 120 piglets. Digesta and tissue samples were taken from the ascending colon after 7 ± 1, 14 ± 1, 21 ± 1, and 28 ± 1 d. Partially stripped tissue was mounted in Ussing chambers, and histamine was applied either to the serosal or mucosal compartments. Tissue was pretreated with or without aminoguanidine and amodiaquine to block the histamine-degrading enzymes diamine oxidase (DAO) and histamine -methyltransferase (HMT), respectively. Gene expression and catalytic activity of DAO and HMT in the tissue were analyzed. The numbers of mast cells were determined in tissue samples, and histamine concentration was measured in the colon digesta. Colon tissue from another 12 piglets was used for functional studies on histamine H and H receptors by using the neuronal conduction blocker tetrodotoxin (TTX) and the H and H receptor blocker chloropyramine and famotidine, respectively. After serosal histamine application to colonic tissue in Ussing chambers, the change of short-circuit current (Δ) was not affected by pretreatment and was not different between Zn feeding groups. The Δ after mucosal histamine application was numerically lower ( = 0.168) in HZn compared to LZn and NZn pigs. Mast cell numbers increased from 32 to 46 d of life ( < 0.05). Further studies elucidated that the serosal histamine response was partly inhibited by chloropyramine or famotidine ( < 0.01). The response to mucosal histamine tended to be decreased when chloropyramine but not famotidine was applied from either the serosal or the mucosal side ( = 0.055). Tetrodotoxin alone or in combination with chloropyramine resulted in a similar reduction in the mucosal histamine response ( < 0.01). In conclusion, the present study could not identify marked changes in colonic histamine metabolism on dietary ZnO oversupplementation. For the first time, however, H receptors were functionally identified in the pig colon that are localized either on neurons or on cells that activate secretion via neurons. Luminal histamine can elicit a secretory-type response via these receptors.

Enhancement of immunohistochemical detection of Salmonella in tissues of experimentally infected pigs.

Salmonella Typhimurium is one of the main pathogens compromising porcine and human health as well as food safety, because it is a prevailing source of foodborne infections due to contaminated pork. A prominent problem in the management of this bacteriosis is the number of subclinically infected carrier pigs. As very little is known concerning the mechanisms allowing Salmonella to persist in pigs, the objective of this study was to develop an immunohistochemical approach for the detection of salmonellae in tissue of pigs experimentally infected with Salmonella Typhimurium. Samples were obtained from a challenge trial in which piglets of the German Landrace were intragastrically infected with Salmonella enterica serovar Typhimurium DT104 (1.4-2.1x1010 CFU). Piglets were sacrificed on days 2 and 28 post infection. Tissue samples of jejunum, ileum, colon, ileocecal mesenteric lymph nodes (Lnn. ileocolici), and tonsils (Tonsilla veli palatini) were fixed in Zamboni's fixative and paraffin-embedded. Different immunohistochemical staining protocols were evaluated. Salmonella was detected in varying amounts in the tissues. Brown iron-containing pigments in the lymph nodes interfered with the identification of Salmonella if DAB was used as a staining reagent. Detergents like Triton X-100 or Saponin enhanced the sensitivity. It seems advisable not to use a detection system with brown staining for bacteria in an experimental setup involving intestinal damage including haemorrhage. The use of detergents appears to result in a higher sensitivity in the immunohistochemical detection of salmonellae.

Intraepithelial lymphocyte numbers and histomorphological parameters in the porcine gut after Enterococcus faecium NCIMB 10415 feeding in a Salmonella Typhimurium challenge.

Salmonellae are among the most widespread sources of foodborne infections and Salmonella Typhimurium, in particular, is correlated with human disease caused by the consumption of contaminated pork. Intraepithelial lymphocytes (IEL) have early contact with intestinal antigens and play an important role in the detection of pathogenic bacteria. The objective of this study was to determine whether a presumed probiotic Enterococcus faecium strain could improve histomorphological and immune system-related parameters of gut function after a Salmonella challenge in weaned pigs. In particular the morphological parameters villus length and width, crypt depth and width as well as the actual enlargement of the intestinal epithelial surface were calculated and the number of IEL was evaluated in sections of the porcine gut. Weaned piglets were challenged with Salmonella enterica serovar Typhimurium DT 104, and half of them also received Enterococcus faecium NCIMB 10415 in the diet. Animals were sacrificed at days post infection (DPI) 2 and 28. The effect of the factors "time post-infection/age" and "probiotic treatment" on jejunal morphology and IEL numbers and distribution was evaluated by light microscopy. The time post-infection had significant effects in both feeding groups. Animals sacrificed at DPI 28 had longer and wider villi, deeper and wider crypts, a higher villus enlargement factor, a higher ratio between villus and crypt enlargement factors as well as more IEL. Probiotic treatment resulted in longer villi, a higher ratio of villus surface/crypt circumference enlargement factors and significantly more IEL. The larger total number of IEL displayed by the probiotic group resulted from significantly higher numbers of IEL at the nuclear and apical levels of the intraepithelial compartment but not from the number of IEL situated at the basement membrane. The probiotic effects were only measurable 28 DPI. It is proposed that Enterococcus faecium NCIMB 10415 exerts an immune modulatory effect by increasing the numbers of intraepithelial lymphocytes.

[Ophthalmological imaging with ultrahigh field magnetic resonance tomography: technical innovations and frontier applications]. / Ophthalmologische Bildgebung mit Ultrahochfeld-Magnetresonanztomografie: technische Innovationen und wegweisende Anwendungen.

This review documents technical progress in ophthalmic magnetic resonance imaging (MRI) at ultrahigh fields (UHF, B(0) ≥ 7.0 T). The review surveys frontier applications of UHF-MRI tailored for high spatial resolution in vivo imaging of the eye, orbit and optic nerve. Early examples of clinical ophthalmic UHF-MRI including the assessment of melanoma of the choroid membrane and the characterisation of intraocular masses are demonstrated. A concluding section ventures a glance beyond the horizon and explores research promises along with future directions of ophthalmic UHF-MRI.

Effect of dietary zinc oxide on jejunal morphological and immunological characteristics in weaned piglets.

The aim of this study was to evaluate age-related changes and the effect of dietary Zn concentration on morphological and immunological characteristics in the gastrointestinal tract of piglets. A total of 96 purebred Landrace piglets were weaned at the age of 26 ± 1 d, and randomly allocated into 3 treatment groups fed with low (57 mg Zn/kg), medium (164 mg Zn/kg), and high (2425 mg Zn/kg) dietary Zn (ZnO). Piglets (4 males and 4 females per treatment group) were killed at 33 ± 1, 40 ± 1, 47 ± 1, and 54 ± 1 d of age. In the jejunum, villus height, crypt depth, and the number of goblet cells producing neutral, acidic, sulfated, and sialylated mucins were measured. Intraepithelial lymphocytes were characterized by flow cytometry and the gene expression of mucin 2 (MUC2), mucin 20 (MUC20), ß-defensin 3, and trefoil factor 3 (TFF3) was determined by reverse transcription quantitative PCR. Villus height and crypt depth in the jejunum showed age related differences (P < 0.01), whereas the dietary concentrations of Zn had no effect. The mucin types were modified mainly by age, and dietary Zn had no effect in the proximal jejunum. In the distal jejunum, age and Zn had effects on the mucin types. The abundance of sulfomucins decreased (P < 0.001) and sialomucins increased with age (P < 0.001), while high dietary ZnO reduced the sulfomucins (P < 0.001) and increased the sialomucins (P < 0.05) in the crypts. The phenotypes of lymphocytes in the epithelium of the proximal jejunum showed relatively constant percentages of T-cells, as well as natural killer cells. High dietary Zn treatment led to a reduced abundance of CD8(+) γδ T-cells (P < 0.05). The apportionment of different cytotoxic T-cell was age dependent. Although the percentage of CD4(-)CD8ß(+) increased (P < 0.01), the relative amount of CD4(+)CD8ß(+) decreased with age (P < 0.05). The expression of MUC2 and MUC20 was not influenced by age or dietary Zn concentration. High Zn intakes resulted in a reduced gene expression of ß-defensin 3 (P < 0.05), but did not affect the expression of TFF3. It is concluded that Zn in the form of ZnO appears to have specific effects on the innate and adaptive gut associated immune system of piglets. These might contribute to the positive effects of Zn on the prevention of postweaning diarrhea.

Characterization of CD4+ subpopulations and CD25+ cells in ileal lymphatic tissue of weaned piglets infected with Salmonella Typhimurium with or without Enterococus faecium feeding.

The aim of the present study was to test the effect of Enterococcus faecium NCIMB 10415 (E. faecium) on CD4+ T helper immune cell subpopulations and CD25+ cells in ileal lymphatic tissue after challenge with Salmonella (S.) Typhimurium DT 104. German Landrace piglets treated with E. faecium (n=16) as a feed additive and untreated controls (n=16) were challenged with S. Typhimurium 10 days after weaning. The expression of lineage specific T helper cell subtype master transcription factors on mRNA level was measured in the whole tissue of the gut associated lymphoid tissues (ileocecal mesenteric lymph node, ileum with Peyer's patches and papilla ilealis) and in magnetically sorted T helper cells from blood and ileocecal mesenteric lymph nodes at two and 28 days post infection. CD25 protein expression of T helper cells was studied by flow cytometry in ileal Peyer's patches, lymph nodes and blood. Distribution and morphology of CD25+ cells was demonstrated in situ by immunohistochemistry in paraffin embedded specimens of the ileum and the ileocecal mesenteric lymph nodes. The data provide evidence for a higher T helper 2 cell driven immune response in the control group compared to the E. faecium treated group (P<0.05) in CD4+ magnetically sorted lymphocytes from the ileocecal mesenteric lymph nodes at two and 28 days post infection. We did not observe differences for CD25+ cells in immunohistochemistry and flow cytometry between E. faecium fed pigs and the control group, but provided a detailed description of the occurrence and morphology of these cells in the gut associate lymphoid tissues of piglets. In conclusion we suggest that (i) prolonged feeding with E. faecium can result in changes of the T helper cell response leading to a stronger infection with S. Typhimurium and (ii) that it is important to examine purified immune cells to be able to detect effects on T helper cell subpopulations.

The relationship between lingual and hypoglossal nerve function and quality of life in head and neck cancer.

Sensorimotor impairment of the tongue has the potential to affect speech and swallowing. The purpose of this study was to critically examine the effects of nerve preservation and reinnervation after reconstruction of the base of tongue on patient-perceived outcomes of quality of life (QoL) related to speech and swallowing through completion of the EORTC QLQ-H&N35 standardised questionnaire. Thirty participants with a diagnosis of base of tongue cancer underwent primary resection and reconstruction with a radial forearm free flap, which may or may not have included nerve repair to the lingual nerve, hypoglossal nerve or both. Eight QoL domains sensitive to changes in motor and sensory nerve function were included in the analysis. Transected lingual and hypoglossal nerves were associated with difficulty in swallowing, social eating, dry mouth and social contact. There were fewer problems reported when these nerves were either repaired or left intact. There were no significant differences between patient nerve status and QoL outcomes for speech, sticky saliva and use of feeding tubes. This study was the first to examine the impact of sensory or motor nerve transection and reconstruction on health-related QoL outcomes.

Porcine intestinal mast cells. Evaluation of different fixatives for histochemical staining techniques considering tissue shrinkage.

Staining of mast cells (MCs), including porcine ones, is critically dependent upon the fixation and staining technique. In the pig, mucosal and submucosal MCs do not stain or stain only faintly after formalin fixation. Some fixation methods are particularly recommended for MC staining, for example the fixation with Carnoy or lead salts. Zinc salt fixation (ZSF) has been reported to work excellently for the preservation of fixation-sensitive antigens. The aim of this study was to establish a reliable histological method for counting of MCs in the porcine intestinum. For this purpose, different tissue fixation and staining methods that also allow potential subsequent immunohistochemical investigations were evaluated in the porcine mucosa, as well as submucosa of small and large intestine. Tissues were fixed in Carnoy, lead acetate, lead nitrate, Zamboni and ZSF and stained subsequently with either polychromatic methylene blue, alcian blue or toluidine blue. For the first time our study reveals that ZSF, a heavy metal fixative, preserves metachromatic staining of porcine MCs. Zamboni fixation was not suitable for histochemical visualization of MCs in the pig intestine. All other tested fixatives were suitable. Alcian blue and toluidine blue co-stained intestinal goblet cells which made a prima facie identification of MCs difficult. The polychromatic methylene blue proved to be the optimal staining. In order to compare MC counting results of the different fixation methods, tissue shrinkage was taken into account. As even the same fixation caused shrinkage-differences between tissue from small and large intestine, different factors for each single fixation and intestinal localization had to be calculated. Tissue shrinkage varied between 19% and 57%, the highest tissue shrinkage was found after fixation with ZSF in the large intestine, the lowest one in the small intestine after lead acetate fixation. Our study emphasizes that MC counting results from data using different fixation techniques can only be compared if the respective study-immanent shrinkage factor has been determined and quantification results are adjusted accordingly.

Validación en castellano de un cuestionario breve útil para cribado epidemiológico de epilepsia en España: Cuestionario EPIBERIA / Validation of a short useful questionnaire in Spanish for the epidemiological screening of epilepsy in Spain. EPIBERIA Questionnaire

Introducción: Existe una importante laguna de conocimiento sobre la epidemiología de la epilepsia en los países de la cuenca mediterránea. El grupo EPIBERIA nace con el objetivo de promocionar la realización de estudios epidemiológicos en este ámbito, capaces de paliar esta situación. El presente trabajo aborda la validación de un cuestionario breve de cribado de pacientes con epilepsia en población general. Métodos: Se seleccionó un cuestionario de origen anglosajón validado en inglés por el grupo de Ottman. Fue traducido, modificado para adaptarlo a las características de la población española y administrado a una muestra de 200 pacientes (93 epilépticos y 107 controles no epilépticos), extraídos de manera consecutiva de 5 unidades de epilepsia dispersas por España. Ambos grupos fueron homogéneos en variables demográficas y el grupo de control fue representativo de la población general. Se realizó una estimación de la sensibilidad (S), la especificidad (E), los valores predictivos positivos (VPP) y los valores predictivos negativos (VPN) para cuatro diferentes criterios de corrección del cuestionario. Resultados: Se obtuvieron una sensibilidad del 100% y una especificidad del 74,77% para el criterio menos riguroso y una sensibilidad del 94,62% y una especificidad del 99,07% para el criterio más estricto de corrección del cuestionario. Los VPP variaron entre el 7,48% en el primer supuesto y el 69,49% en el segundo, asumiendo una prevalencia pretest para la epilepsia del 2%. Conclusiones: El cuestionario EPIBERIA es un instrumento válido como cuestionario de cribado de epilepsia en la población general en castellano en España(AU)

Introduction: There is a major gap in knowledge about the epidemiology of epilepsy in Mediterranean countries. The EPIBERIA group was formed with the aim of promoting the conducting of epidemiological studies in this area in order to improve this situation. This paper deals with the validation of a brief questionnaire for screening of patients with epilepsy in general population. Methods: We selected an English language questionnaire previously validated by the Ottman group. It was translated, modified to suit the characteristics of the Spanish population, and administered to a sample of 200 patients (93 epileptics and 107 non-epileptic patient controls) sampled consecutively from 5 Epilepsy Units scattered throughout Spain. Both groups were homogeneous in demographic variables, and the control group was representative of the general population. Results: We obtained a sensitivity of 100% and a specificity of 74.77% for the less rigorous correction criteria of the questionnaire, with a sensitivity of 94.62% and a specificity of 99.07% for the most stringent ones. The positive predictive values (PPVs) ranged from 7.48% for the first case to 69.49% in the second, assuming a prevalence for epilepsy of 2%. Conclusions: The questionnaire EPIBERIA is a valid Spanish tool for epilepsy screening of epilepsy in the general population in Spain(AU)

Validation of a short useful questionnaire in Spanish for the epidemiological screening of epilepsy in Spain. EPIBERIA Questionnaire.

INTRODUCTION: There is a major gap in knowledge about the epidemiology of epilepsy in Mediterranean countries. The EPIBERIA group was formed with the aim of promoting the conducting of epidemiological studies in this region in order to improve this situation. This paper deals with the validation of a brief questionnaire for screening patients with epilepsy in the general population. METHODS: We selected an English-language questionnaire previously validated by the Ottman group. It was translated, modified to suit the characteristics of the Spanish population, and administered to a sample of 200 patients (93 epileptics and 107 non-epileptic patient controls) sampled consecutively from 5 epilepsy units in different cities in Spain. Both groups were homogeneous in demographic variables and the control group was representative of the general population. RESULTS: We obtained a sensitivity of 100% and a specificity of 74.77% for the least rigorous correction model for the questionnaire, with a sensitivity of 94.62% and a specificity of 99.07% for the most stringent correction model. The PPV ranged from 7.48% for the first case to 69.49% in the second, assuming an epilepsy prevalence of 2%. CONCLUSIONS: The questionnaire EPIBERIA is a valid Spanish tool for epilepsy screening in the general population in Spain.