Use and meaning of the anatomical terms plexus choroideus and tela choroidea in veterinary and human medicine.

The anatomical terms plexus chor(i)oideus (CP) and tela chor(i)oidea (TC) are listed without explanations in the official nomenclature handbooks Terminologia Neuroanatomica and Nomina Anatomica Veterinaria. Definitions of CP and TC exhibit discrepancies in medical dictionaries and anatomy handbooks. The aim of our study was to analyse this problem in detail and to discuss a possible unified use of the terms in science and teaching. We conducted a systematic literature review based on the Preferred Reporting Items for Systematic Reviews and Meta-Analyses, identifying and analysing relevant scholarly articles. Additionally, comprehensive original handbooks on human and veterinary anatomy in English and other European languages were examined. The definitions of the terms CP and TC differed considerably between articles and did not match the most frequently given explanations in handbooks. In general use, it seems to have become accepted that TC represents the smooth, thin part of the roof of third and fourth ventricles, and CP the frond- or fringe-like vascularised structures invaginated into lateral, third and fourth ventricles. However, it is controversial which tissue layers should be included in their description. Etymologically, only the vascular network should be termed (choroid) plexus, but embryologically and functionally, epithelium, pial connective tissue and vascular network form an inseparable entity. Similarly, the smooth part of the ventricle roof consists of a (less) vascularised pia-derived stroma and lining epithelium. Including all these layers in CP as well as TC definition might be advisable and also corresponds to the use of the terms in clinical context.

Periphery of porcine hepatic lobes has the smallest length density of hepatic sinusoids and bile canaliculi: A stereological histological study with implications for liver biopsies.

BACKGROUND: Porcine liver is widely used in hepatologic research as a large animal model with many anatomical and physiological similarities with humans. However, only limited information on porcine liver spatial microstructure has been published, especially regarding the hepatic sinusoids and bile canaliculi. The aim of our study was to quantify the sinusoidal and bile canalicular network in healthy male and female porcine livers and to map the variability of these structures with heterogenous distribution to improve the evaluability of liver biopsy samples. METHODS: Livers from 12 healthy piglets (6 females and 6 neutered males) were sampled into 36 tissue samples per organ, representing six hepatic lobes and three different regions related to the hepatic vasculature (peripheral, paracaval and paraportal region). Histological sections were processed with a random orientation of the cutting plane. The endothelium and the bile canaliculi were stained using Ricinus communis agglutinin I lectin histochemistry. The length densities of hepatic sinusoids LV(sinusoids,liver), of bile canaliculi LV(bile canaliculi,liver) and volume fraction VV(sinusoids,liver) and surface density SV(sinusoids,liver) of sinusoids were estimated using stereological methods. The newly acquired morphometric data were compared with previously published data on density of porcine hepatocytes and fractions of connective tissue. RESULTS: The peripheral region had smallest LV(sinusoids,liver), smallest LV(bile canaliculi,liver) and greatest VV(sinusoids,liver). The six hepatic lobes had statistically comparable length densities of both sinusoids and bile canaliculi, but the left lateral lobe had smallest VV(sinusoids,liver). Regions with greater LV(sinusoids,liver) had also greater LV(bile canaliculi,liver) and SV(sinusoids,liver) and were accompanied by greater density of smaller hepatocytes. Regions with smaller LV(sinusoids,liver) and LV(bile canaliculi,liver) contained a greater fraction of interlobular connective tissue. CONCLUSIONS: The length density of hepatic sinusoids is smaller in the peripheral regions of the porcine liver than in other regions related to the hepatic vasculature - paracaval and paraportal regions, and smaller in castrated males than in females. Greater length density of liver sinusoids was linked with greater local density of bile canaliculi, with local increase in the density of smaller hepatocytes and, simultaneously, with smaller fractions of hepatic connective tissue. The intrahepatic and inter-sexual variability of the porcine liver morphology needs to be taken into account when designing and interpreting experiments involving the histological quantification of the microvascular network. The complete primary morphometric data describing the distribution of morphometric parameters within porcine liver were made available in a form facilitating the power analysis to justify the minimal number of tissue samples or animals required when designing further histological evaluation studies. The macroscopic map of microvessels and bile canaliculi variability facilitates their assessment in liver biopsies in the pig.

Comparison of Six Different Methods for Measuring the Equine Hoof and Recording of its Three-Dimensional Conformation.

Different measuring techniques have been used to objectify the classification of hoof shape. The MicroScribe is a novel tool that might prove useful for measuring hooves without prior reconstruction or compensation of projection artefacts. The aim of this study was to compare biometric data of the equine hoof collected by the MicroScribe tool and measurements collected directly from hooves, scaled photographs and radiographs, from photogrammetry models and computed tomography datasets. The suitability of MicroScribe generated data to differentiate individual hoof conformations was tested. A total of 62 measures were recorded from 16 forehooves. 21 linear and nine angular measures were collected by at least four methods each, and evaluated further by analysis of variance (ANOVA) and multivariate analysis of variance (MANOVA). Ratios and differences of these measures were calculated as suitable for the definition of hoof shapes and analysed as well. Absolute equivalency of methods was detected for five linear and none of the angular measurements. The precision of the tested measurement methods was comparable. In some cases, different methods measure different structures. Radiographs tended to overestimate, while computed tomography slides to underestimate distances. Photogrammetry and scaled photographs were less suitable for measuring hoof angles. The MicroScribe tool can readily be used for hoof measurements. Its values for linear measures showed good equivalency with other methods based on real hooves. For angular measurements, the uneven hoof surface might introduce imprecision. Not all hoof conformations could be detected based on measuring results alone. Diagnosis by a skilled veterinarian is still essential.

Using virtual microscopy for the development of sampling strategies in quantitative histology and design-based stereology.

Only a fraction of specimens under study are usually selected for quantification in histology. Multilevel sampling or tissue probes, slides and fields of view (FOVs) in the regions of interest (ROIs) are required. In general, all parts of the organs under study should be given the same probability to be taken into account; that is, the sampling should be unbiased on all levels. The objective of our study was to provide an overview of the use of virtual microscopy in the context of developing sampling strategies of FOVs for stereological quantification. We elaborated this idea on 18 examples from multiple fields of histology, including quantification of extracellular matrix and muscle tissue, quantification of organ and tumour microvessels and tumour-infiltrating lymphocytes, assessing osseointegration of bone implants, healing of intestine anastomoses and osteochondral defects, counting brain neurons, counting nuclei in vitro cell cultures and others. We provided practical implications for the most common situations, such as exhaustive sampling of ROIs, sampling ROIs of different sizes, sampling the same ROIs for multiple histological methods, sampling more ROIs with variable intensities or using various objectives, multistage sampling and virtual sampling. Recommendations were provided for pilot studies on systematic uniform random sampling of FOVs as a part of optimizing the efficiency of histological quantification to prevent over- or undersampling. We critically discussed the pros and cons of using virtual sections for sampling FOVs from whole scanned sections. Our review demonstrated that whole slide scans of histological sections facilitate the design of sampling strategies for quantitative histology.

Endodontic Treatment of a Traumatic Tusk Fracture With Exposed Pulp in an Asian Elephant (Elephas maximus).

Tusk fracture in elephants is a common incident often resulting in pulp exposure and pulpitis. Extensive lavage, endodontic therapy, direct pulp capping, or extraction are treatment options. In this report, the successful management of a broken tusk of a juvenile male Asian elephant (Elephas maximus) including morphological analysis of the tusk tip 2 years after surgery are presented. Treatment was carried out under barn conditions and included antimicrobial photodynamic therapy and partial pulpotomy with direct pulp capping. Immediate pain relief was reached. The fractured tusk was preserved and continued to grow. The therapeutic filling material remained intact for over 1 year but was absent 2 years after treatment. The former pulp cavity of the tusk tip was filled with reparative dentin, osteodentin, and bone, but the seal between these hard tissues and pulp chamber dentin was incomplete. Radiographs obtained 3 years after treatment showed no differences in pulp shape, pulp width, and secondary dentin formation between the treated right and the healthy left tusk. It can be concluded that in case of an emergency, the endodontic therapy of a broken elephant tusk can be attempted under improvised conditions with adequate success. Photodynamic therapy might contribute to prevent infection and inflammation of the pulp. The decision tree published by Steenkamp (2019) provides a valuable tool to make quick decisions regarding a suitable therapy of broken tusks.

Regional Collagen Fiber Network in the Articular Disc of the Human Temporomandibular Joint: Biochemical 3-Tesla Quantitative Magnetic Resonance Imaging Compared to Quantitative Histologic Analysis of Fiber Arrangement.

AIMS: To evaluate the regional collagen fiber network in the human temporomandibular joint (TMJ) disc by using biochemical magnetic resonance imaging (MRI) and quantitative histology. METHODS: MRI of 5 heads (10 TMJ discs) obtained from partially dentate or edentulous cadavers was performed at 3-Tesla MRI by using a flexible, 8-channel transmit-receive coil. After MRI, all 10 discs were processed histologically. Percentages of coronal, sagittal, and transverse collagen fibers were assessed stereologically for the anterior, central, and posterior parts of the disc. An anisotropy index was calculated for collagen fiber arrangement in all three regions of interest. RESULTS: In the central part of the TMJ disc, collagen fibers were arranged anisotropically with a preferentially sagittal direction. In the anterior and posterior parts, evidence for fibers being arranged isotropically (randomly) without preferred direction was found. Mean MRI T2 values appeared to be correlated with the anisotropy index of collagen fibers (r = -0.45; P < .05). When tested individually, T2 values of the isotropic anterior and posterior disc regions showed a partial but significant correlation with the anisotropy index of collagen fibers (r = -0.54; P < .05), whereas the anisotropic central part did not (P > .05). CONCLUSION: This study has provided the first systematic comparison of quantitative data on collagen fiber isotropy and anisotropy assessed in histologic sections with biochemical quantitative MRI for human TMJ fibrous cartilage.

[Untersuchung des Tränenfilms und histologische Untersuchung der Augenlider von Möpsen]. / Tear film examinations and eyelid histology of pugs.

OBJECTIVES: To determine if qualitative tear film and histological changes of the eyelid margins in pugs compared to other brachycephalic dogs could be potential contributing factors to the high prevalence of corneal diseases in this breed. METHODS: Ophthalmic examin ation (including tear film break-up time [TFBUT] and meibometry) was undertaken on three groups (pugs with and without ophthalmologic abnormalities as well as on other brachycephalic breeds with history of ophthalmologic abnormalities). Histology of eyelid tissue obtained during medial canthoplasty was performed, using hematoxylin-eosin and oil-red-O-staining. RESULTS: Seventy-eight pugs and 11 brachycephalic dogs were included. Mean ages were 3.54 and 5.5 years respectively. STT 1 values below 15 mm/min were found in 12 of 150 eyes of pugs and in three of 18 eyes of other brachycephalic dogs. Tear film break-up time values below 20 seconds were determined in 118 of 126 eyes of pugs, and in eight of 18 eyes of other brachycephalic dogs. Meibometry values over 100 MU were only identified in 20 of 147 eyes of pugs and 12 of 20 eyes in other brachycephalic dogs. Eyelid tissue of 21 pugs and 11 brachycephalic dogs was obtained. All pugs had a higher number of inflammatory cells in eyelid tissue than other brachycephalic breeds. CONCLUSIONS: Young pugs are often presented with STT 1 values within the reference range and low TFBUT and meibometry values. As other brachycephalic breeds are often presented with STT 1 values within reference ranges as well as low TFBUT values, the only marked difference between pugs and other brachycephalic breeds are low meibometry values and the higher number of inflammatory cells in the medial canthal lid margins.

Numerical and length densities of microvessels in the human brain: Correlation with preferential orientation of microvessels in the cerebral cortex, subcortical grey matter and white matter, pons and cerebellum.

To provide basic data on the local differences in density of microvessels between various parts of the human brain, including representative grey and white matter structures of the cerebral hemispheres, the brain stem and the cerebellum, we quantified the numerical density NV and the length density LV of microvessels in two human brains. We aimed to correlate the density of microvessels with previously published data on their preferential orientation (anisotropy). Microvessels were identified using immunohistochemistry for laminin in 32 samples harvested from the following brain regions of two adult individuals: the cortex of the telencephalon supplied by the anterior, middle, and posterior cerebral artery; the basal ganglia (putamen and globus pallidus); the thalamus; the subcortical white matter of the telencephalon; the internal capsule; the pons; the cerebellar cortex; and the cerebellar white matter. NV was calculated from the number of vascular branching points and their valence, which were assessed using the optical disector in 20-µm-thick sections. LV was estimated using counting frames applied to routine sections with randomized cutting planes. After correction for shrinkage, NV in the cerebral cortex was 1311±326mm-3 (mean±SD) and LV was 255±119mm-2. Similarly, in subcortical grey matter (which included the basal ganglia and thalamus), NV was 1350±445mm-3 and LV was 328±117mm-2. The vascular networks of cortical and subcortical grey matter were comparable. Their densities were greater than in the white matter, with NV=222±147mm-3 and LV=160±96mm-2. NV was moderately correlated with LV. In parts of brain with greater NV, blood vessels lacked a preferential orientation. Our data were in agreement with other studies on microvessel density focused on specific brain regions, but showed a greater variability, thus mapping the basic differences among various parts of brain. To facilitate the planning of other studies on brain vascularity and to support the development of computational models of human brain circulation based on real microvascular morphology; stereological data in form of continuous variables are made available as supplements.

Tropism, intracerebral distribution, and transduction efficiency of HIV- and SIV-based lentiviral vectors after injection into the mouse brain: a qualitative and quantitative in vivo study.

Lentiviruses are suitable to transfer potential therapeutic genes into non-replicating cells such as neurons, but systematic in vivo studies on transduction of neural cells within the complete brain are missing. We analysed the distribution of transduced cells with respect to brain structure, virus tropism, numbers of transduced neurons per brain, and influence of the Vpx or Vpr accessory proteins after injection of vectors based on SIVsmmPBj, HIV-2, and HIV-1 lentiviruses into the right striatum of the mouse brain. Transduced cells were found ipsilaterally around the injection canal, in corpus striatum and along corpus callosum, irrespective of the vector type. All vectors except HIV-2SEW transduced also single cells in the olfactory bulb, hippocampus, and cerebellum. Vector HIV-2SEW was the most neuron specific. However, vectors PBjSEW and HIV-1SEW transduced more neurons per brain (means 41,299 and 32,309) than HIV-2SEW (16,102). In the presence of Vpx/Vpr proteins, HIV-2SEW(Vpx) and HIV-1SEW(Vpr) showed higher overall transduction efficiencies (30,696 and 27,947 neurons per brain) than PBjSEW(Vpx) (6636). The distances of transduced cells from the injection canal did not differ among the viruses but correlated positively with the numbers of transduced neurons. The presence of Vpx/Vpr did not increase the numbers of transduced neurons. Parental virus type and the vector equipment seem to influence cellular tropism and transduction efficiency. Thus, precision of injection and choice of virus pseudotype are not sufficient when targeted lentiviral vector transduction of a defined brain cell population is required.

Stereological analysis of size and density of hepatocytes in the porcine liver.

The porcine liver is frequently used as a large animal model for verification of surgical techniques, as well as experimental therapies. Often, a histological evaluation is required that include measurements of the size, nuclearity or density of hepatocytes. Our aims were to assess the mean number-weighted volume of hepatocytes, the numerical density of hepatocytes, and the fraction of binuclear hepatocytes (BnHEP) in the porcine liver, and compare the distribution of these parameters among hepatic lobes and macroscopic regions of interest (ROIs) with different positions related to the liver vasculature. Using disector and nucleator as design-based stereological methods, the morphometry of hepatocytes was quantified in seven healthy piglets. The samples were obtained from all six hepatic lobes and three ROIs (peripheral, paracaval and paraportal) within each lobe. Histological sections (thickness 16 µm) of formalin-fixed paraffin-embedded material were stained with the periodic acid-Schiff reaction to indicate the cell outlines and were assessed in a series of 3-µm-thick optical sections. The mean number-weighted volume of mononuclear hepatocytes (MnHEP) in all samples was 3670 ± 805 µm3 (mean ± SD). The mean number-weighted volume of BnHEP was 7050 ± 2550 µm3 . The fraction of BnHEP was 4 ± 2%. The numerical density of all hepatocytes was 146 997 ± 15 738 cells mm-3 of liver parenchyma. The porcine hepatic lobes contained hepatocytes of a comparable size, nuclearity and density. No significant differences were identified between the lobes. The peripheral ROIs of the hepatic lobes contained the largest MnHEP with the smallest numerical density. The distribution of a larger MnHEP was correlated with a larger volume of BnHEP and a smaller numerical density of all hepatocytes. Practical recommendations for designing studies that involve stereological evaluations of the size, nuclearity and density of hepatocytes in porcine liver are provided.

Ubiquitous LEA29Y Expression Blocks T Cell Co-Stimulation but Permits Sexual Reproduction in Genetically Modified Pigs.

We have successfully established and characterized a genetically modified pig line with ubiquitous expression of LEA29Y, a human CTLA4-Ig derivate. LEA29Y binds human B7.1/CD80 and B7.2/CD86 with high affinity and is thus a potent inhibitor of T cell co-stimulation via this pathway. We have characterized the expression pattern and the biological function of the transgene as well as its impact on the porcine immune system and have evaluated the potential of these transgenic pigs to propagate via assisted breeding methods. The analysis of LEA29Y expression in serum and multiple organs of CAG-LEA transgenic pigs revealed that these animals produce a biologically active transgenic product at a considerable level. They present with an immune system affected by transgene expression, but can be maintained until sexual maturity and propagated by assisted reproduction techniques. Based on previous experience with pancreatic islets expressing LEA29Y, tissues from CAG-LEA29Y transgenic pigs should be protected against rejection by human T cells. Furthermore, their immune-compromised phenotype makes CAG-LEA29Y transgenic pigs an interesting large animal model for testing human cell therapies and will provide an important tool for further clarifying the LEA29Y mode of action.

Vasa vasorum in the tunica media and tunica adventitia of the porcine aorta.

Vasa vasorum supply both the tunica adventitia and the tunica media of major arteries with nutrients and oxygen. We estimated the density of von Willebrand factor-positive profiles of vasa vasorum visible in transversal histological sections of 123 tissue samples collected from five anatomical positions in the porcine aortae of growing pigs (n=25). The animals ranged in age from 0 to 230 days. The tunica media of the thoracic aorta had a greater vasa vasorum density, with microvessels penetrating deeper towards the lumen than in the abdominal aorta. The density of vasa vasorum gradually decreased with age in both the media and the adventitia. The relative depth into which the vasa vasorum penetrated and where they branched remained constant during the ageing and growth of the media. The ratio of the tunica media and tunica adventitia thicknesses did not change in the single aortic segments during ageing. The media of older animals received fewer but equally distributed vasa vasorum. A greater density of vasa vasorum in the media was correlated with greater media thickness and a greater elastin fraction (data on elastin taken from another study on the same samples). Immunohistochemical quantification revealed deeper penetration of vasa vasorum towards the adluminal layers of the tunica media that were hitherto reported to be avascular. The complete primary morphometric data, in the form of continuous variables, have been made available as a supplement. Mapping of the vasa vasorum profile density and position has promising illustrative potential for studies on atherosclerotic and inflammatory neovascularization, aortic aneurysms, and drug distribution from arterial stents in experimental porcine models.

Segmental and age differences in the elastin network, collagen, and smooth muscle phenotype in the tunica media of the porcine aorta.

The porcine aorta is often used in studies on morphology, pathology, transplantation surgery, vascular and endovascular surgery, and biomechanics of the large arteries. Using quantitative histology and stereology, we estimated the area fraction of elastin, collagen, alpha-smooth muscle actin, vimentin, and desmin within the tunica media in 123 tissue samples collected from five segments (thoracic ascending aorta; aortic arch; thoracic descending aorta; suprarenal abdominal aorta; and infrarenal abdominal aorta) of porcine aortae from growing domestic pigs (n=25), ranging in age from 0 to 230 days. The descending thoracic aorta had the greatest elastin fraction, which decreased proximally toward the aortic arch as well as distally toward the abdominal aorta. Abdominal aortic segments had the highest fraction of actin, desmin, and vimentin positivity and all of these vascular smooth muscle markers were lower in the thoracic aortic segments. No quantitative differences were found when comparing the suprarenal abdominal segments with the infrarenal abdominal segments. The area fraction of actin within the media was comparable in all age groups and it was proportional to the postnatal growth. Thicker aortic segments had more elastin and collagen with fewer contractile cells. The collagen fraction decreased from ascending aorta and aortic arch toward the descending aorta. By revealing the variability of the quantitative composition of the porcine aorta, the results are suitable for planning experiments with the porcine aorta as a model, i.e. power test analyses and estimating the number of samples necessary to achieving a desirable level of precision. The complete primary morphometric data, in the form of continuous variables, are made publicly available for biomechanical modeling of site-dependent distensibility and compliance of the porcine aorta.

Segmental differences in the orientation of smooth muscle cells in the tunica media of porcine aortae.

The orientation of vascular smooth muscle cells of porcine aortae was assessed to test the widely accepted assumption that these smooth muscle cells are arranged in two helices. We used tangential histological sections of 82 samples of five anatomical segments of thoracic and abdominal porcine aortae and three age groups in animals ranging in age from 5 to 210 days. The distribution of the orientation of smooth muscle cell nuclei in five proximodistal segments of the porcine aortae was determined using an algorithm that fitted a mixture of one to five von Mises probability distributions of the data retrieved from histological micrographs. Automated tracking of the nuclei was confirmed by and consistent with manual histological analysis. The orientation of the vascular smooth muscle cells was successfully fitted using two von Mises distributions in most of the samples with different ages, wall thicknesses, and anatomical positions, which corresponds to two populations of vascular smooth muscle cells. A minor fraction of samples also required a tertiary von Mises distribution to describe the orientation of the smooth muscle cell nuclei. The distribution of vascular smooth muscle cells in five aortic segments ranging from the thoracic ascending aorta to the abdominal intrarenal aorta exhibited similar main directions but different shapes. These results are consistent with the widely used model of two muscular helices intermingling in the arterial wall. Furthermore, we calculated the central angles of symmetry and the mean value of angles between the two assumed smooth muscle directions. We also successfully approximated the orientation of the smooth muscle cells using a mixture of von Mises distributions and our open-source software named dist_mixtures. This method is openly available to researchers who are interested in mathematically assessing the orientation of cell nuclei in various tissues.

Distribution of orientation of smooth muscle bundles does not change along human great and small varicose veins.

Wall remodeling in varicose veins is associated with hypertrophy of subendothelial tissue, increase in inner diameter, wrinkling and invagination of the endothelial layer. Due to structural alterations of the wall, the smooth muscle cells (SMCs) change their original circular and longitudinal orientations. Our aim was to quantify the volume fraction of circularly, longitudinally and obliquely oriented SMCs within both the inner and outer half of the wall of 11 great saphenous varicose veins and five small saphenous varicose veins. Using stereological methods applied on cross-sections of the vessels regularly gained each 5 cm along the vessel we determined the wall thickness (846 ± 319 µm, mean ± standard deviation), the volume fraction of circular SMCs in the inner (0.19 ± 0.13) and outer (0.06 ± 0.06) layers, the volume fraction of longitudinal SMCs in the inner (0.06 ± 0.05) and outer (0.05 ± 0.04) layers, the volume fraction of oblique SMCs in the inner (0.15 ± 0.08) and outer (0.09 ± 0.08) layers, and the total volume fraction of SMCs in the inner (0.4 ± 0.1) and outer (0.21 ± 0.09) layers. The volume fraction of SMCs with circular and oblique but not with longitudinal orientation was greater in the inner layer compared to the outer layer. The SMC orientation distribution was uniform along the varicose saphenous veins. With increasing wall thickness, the volume fraction of longitudinal and oblique SMC bundles increased in both layers at the expansion of circular SMC bundles. The main differences in the orientation of the SMCs in the inner and outer wall layers should be taken into account when computational modeling of varicose saphenous veins is attempted.

Isospora suis in an epithelial cell culture system - an in vitro model for sexual development in coccidia.

Coccidian parasites are of major importance in animal production, public health and food safety. The most frequently used representative in basic research on this group is Toxoplasma gondii. Although this parasite is well investigated there is no adequate in vitro model for its sexual development available and knowledge on this important life cycle phase is therefore scarce. The use of Isosporasuis, a sister taxon to T. gondii and the causative agent of piglet coccidiosis, could provide a solution for this. In the present study an in vitro model for neonatal porcine coccidiosis in cells representative for the in vivo situation in the piglet gut was developed and evaluated. The parasite development was investigated by light and transmission electron microscopy and optimum culture conditions were evaluated. Intestinal porcine epithelial cells (IPEC-J2) adequately representing the natural host cells supported the development of all endogenous life cycle stages of I. suis, including gametocytes and oocysts. A concentration of 5% fetal calf serum in the culture medium led to highest gametocyte densities on day 12 post infection. Low infection doses (≤1 sporozoite for 100 host cells) were best for oocyst and gametocyte development. The presented system can also be used for immunostaining with established antibodies developed against T. gondii (in our case, anti-TgIMC3 antibodies directed against the inner membrane complex 3). The complete life cycle of I. suis in a cell line representing the natural host cell type and species provides a unique model among coccidian parasites and can be used to address a wide range of topics, especially with regard to the sexual development of coccidia.

Asymptomatic abdominal aortic aneurysms show histological signs of progression: a quantitative histochemical analysis.

OBJECTIVE: Abdominal aortic aneurysm (AAA) is a serious disease due to its covert nature, relatively high prevalence and fatal prognosis in the case of rupture. To obtain new insights into AAA pathogenesis, we examined the relationships between histopathology, multiplex in vitro immunoassay data, diameter and symptomatology. METHODS: In a prospective, non-randomised study, we evaluated samples from 6 normal infrarenal aortae and 65 AAA patients (65 walls, 55 thrombi). The AAA patients were either asymptomatic (n = 44), symptomatic (n = 7) or with ruptured AAA (n = 14). The AAA diameter was classified as small (<5 cm, n = 18), medium (5-7 cm, n = 26) and large (>7 cm, n = 21). We quantified the histopathology of the AAA wall and the adjacent thrombus. We assessed the expression of proteins in the same samples. RESULTS: Asymptomatic AAAs had walls with more abundant inflammatory infiltrates, lower amounts of PAI-1, a higher number of tPA-positive elements, a tendency towards decreased collagen content, whereas the adjacent thrombi had a greater concentration of VCAM-1 and MMP-2 when compared with symptomatic AAAs. Compared with the aneurysmatic aorta, the normal aorta contained less collagen and more elastin, actin, desmin and PAI-1-positive elements; in addition, it was more vascular. Medium-sized AAAs were the most actin and vimentin rich, and large AAAs were the most vascular. CONCLUSION: Our results show that asymptomatic AAA walls often have more potentially deleterious histopathological alterations than symptomatic AAA walls. This result indicates that a progression from an asymptomatic AAA to rupture can be expected and screening patients who are at risk of rupture could be beneficial.

Porcine CD27: identification, expression and functional aspects in lymphocyte subsets in swine.

Up to now for Swine Workshop Cluster 2 (SWC2) the orthologous human CD molecule was unknown. By use of the SWC2-specific mAb b30c7 and a retroviral cDNA expression library derived from stimulated porcine peripheral blood mononuclear cells we could identify SWC2 as porcine CD27. Phenotypic analyses of lymphocytes isolated from blood and lymphatic organs revealed that mature T cells in thymus and T cells in the periphery with a naïve phenotype were CD27(+). However, within CD8α(+) T helper and CD8α(+) γδ T cells also CD27(-) cells were present, indicating a down-regulation after antigen contact in vivo. B cells lacked CD27 expression, whereas NK cells expressed intermediate levels. Furthermore, plate-bound mAb b30c7 showed a costimulatory capacity on CD3-activated T cells for proliferation, IFN-γ and TNF-α production. Hence, our data indicate an important role of porcine CD27 for T-cell differentiation and activation as described for humans and mice.

Vasa vasorum quantification in human varicose great and small saphenous veins.

Recent research regarding saphenous vasa vasorum (VV) has focused on two main topics: the VV during varicogenesis in chronic venous insufficiency and the VV in saphenous grafts used in reconstructive vascular surgery. Our aim has been (i) to establish a technique for the histological quantification of the VV in human varicose great and small saphenous veins and (ii) to describe the density and distribution of the vasa vasorum within varicose veins. Great (n=11) and small (n=5) saphenous veins (length, 15-40cm) were collected from 12 patients who were undergoing venous stripping due to chronic venous insufficiency (Clinical-Etiology-Anatomy-Pathophysiology class 2-3). The veins were divided into 5-cm long segments. In total, 92 tissue blocks were collected to trace the variability of the density and distribution of the vasa vasorum in the proximo-distal direction. The endothelium was detected by immunohistochemistry using the von Willebrand factor. We quantified the number of microvessel profiles per section area and the relative distance of the microvessels from the outer border of the adventitia. The VV did not exhibit a preferential orientation in the varicose veins. VV density profiles were highest in the middle third of the venous wall and lowest in the inner third of the venous wall. Both the density and distribution of VV were uniform along the veins, and no differences were observed between the great and small saphenous veins. The VV density was statistically independent of the relative distance from the adventitia. The usability of this technique for perioperative frozen sections remains to be tested.

Immunohistochemical detection and quantification of T cells in the small intestine of Isospora suis-infected piglets--influence of fixation technique and intestinal segment.

Quantification of immunohistochemical results constitutes an important tool in the analysis of cells and tissue that is not readily replaced by other techniques. For reliable quantification, it is essential to consider factors such as tissue fixation and tissue sampling. We report a study on the model of the intestine of Isospora suis-infected piglets, in which we addressed (1) whether the quantity of detectable T cells in the intestinal mucosa is the same in formalin-, HOPE®-, and cryo-conserved material or whether the amounts of T cells at least correlate with one another; and (2) whether single jejunal segments differ in regard to the quantity of mucosal T cells and variability of lymphocyte infiltration. Quantification of T cells in histological sections of different parts of the jejunum of 15-22 day old piglets infected with I. suis was performed using an anti-CD3-antibody and stereological point counting. Area fractions of T-cell profiles per intestinal mucosa profile were higher in cryo-conserved samples than in HOPE®- and formalin-conserved material but no correlation between different fixations could be found. The proximal part of the jejunum contained fewer T cells compared with mid- and end-jejunum. Coefficients of variation did not differ between the intestinal segments. For quantification of T cells in the gut mucosa of piglets infected with I. suis, the cryo-conserved mid jejunum seems most suitable in cases when unbiased sampling of the complete intestine is not feasible. It is generally not possible to compare quantitative results of immunostaining in samples conserved by different methods.