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Background: Facial emotion recognition (FER) may be impaired in patients with multiple sclerosis (MS). Nevertheless, the literature is heterogeneous, with studies not highlighting this kind of impairment. Moreover, most studies have not explored differences between MS spectrum disorders (radiologically isolated syndrome (RIS), clinically-isolated syndrome (CIS), relapsing-remitting (RRMS), and progressive (primary - (PPMS) and secondary - (SPMS)). One hypothesis would be that FER impairment results from an alteration of eye-gaze strategies while observing emotional faces. Consequently, a FER deficit would be found in MS patients for whom these observation strategies would be disturbed and more frequent in the progressive forms., Methods: We prospectively enroled 52 patients (10 RIS, 10 CIS, 12RRMS, 10 SPMS, 10 PPMS) and 23 healthy controls (HC) to assess FER using Ekman Faces Test. Eye movements (number and duration of fixations) were recorded with an eye-tracking device., Results: 21% of the MS participants had significant FER impairment. This impairment was observed in all phenotypes. In progressive forms, FER impairment was more frequent, more severe, and associated with modified emotional face observation strategies. MS participants with significant FER impairment had significantly more modification of eye-gaze strategies during observation of expressive faces than MS participants without FER impairment., Conclusion: FER impairment seems to be linked to a deficit of attention orientation in MS. Remediation of eye-gaze strategies during observation of emotional faces could be beneficial, as observed in other neurological diseases., Competing Interests: Declaration of Competing Interest None., (Copyright © 2022 Elsevier B.V. All rights reserved.)

This report describe the first application of environmental DNA-metabarcoding approach for the assessment of fish species diversity in two marine protected areas of the North Sea: the Doggerbank and the Sylt Outer Reef. We collected 64 water samples and detected 24 fish species. We discuss qualitative differences between MPAs and compare the results with those obtained from bottom-trawl surveys in the same areas. We found three additional species to those documented in the same year with trawls, including the critically endangered European eel., (© 2022 Fisheries Society of the British Isles.)

Background: Facial emotion recognition (FER) is primarily and severely impaired in individuals with the behavioral variant of frontotemporal dementia (bvFTD) and is often mildy impaired in individuals with Alzheimer disease (AD) or Parkinson disease (PD). Such impairment is associated with inappropriate social behaviors., Objective: To determine whether FER impairment is linked to the use of inappropriate eye-gaze strategies to decode facial emotions, leading to misinterpretation of others' intentions and then to behavioral disorders., Method: We assessed FER in 9 individuals with bvFTD, 23 with AD, and 20 with PD, as well as 22 healthy controls (HC), using the Reading the Mind in the Eyes (RME) Test and the Ekman Faces Test. Eye movements (number and duration of fixations) were recorded with an eye-tracking device. Behavior was assessed using the Neuropsychiatric Inventory., Results: FER was mildly impaired in the AD and PD groups and severely impaired in the bvFTD group. FER impairment was accompanied by an increase in the number of fixations and a more attracted gaze toward the lower part of one's face. FER impairment and an increase in the number of fixations were positively correlated with behavioral disorders., Conclusion: Our study demonstrated a link between FER impairment, modification of eye-gaze strategies during the observation of emotional faces, and behavioral disorders in individuals with bvFTD and those with AD or PD. These results suggest that an eye-gaze strategy rehabilitation program could have beneficial effects on emotion recognition and behavioral disorders in individuals with these diseases., Competing Interests: The authors declare no conflicts of interest., (Copyright © 2022 Wolters Kluwer Health, Inc. All rights reserved.)

Objective: Theory of mind (ToM) and empathy are severely impaired in the behavioral-variant of frontotemporal dementia (bvFTD) and more mildly in Alzheimer's (AD) and Parkinson's diseases (PD). Such impairments are associated with behavioral disorders (BD). Modification of visual scanning strategies of complex visual scenes is also found in these pathologies. We hypothesized that these patients applied atypical gaze strategies when observing social events, which would not allow to properly process social cues and would result in the production of erroneous inferences and lack of empathy towards others., Methods: Fifty-five participants were divided into four groups: five bvFTD, 19 AD, 17 PD and 14 matched controls subjects. ToM and empathy were assessed by eye movements recording (eye-tracking) and by a questionnaire during a painting observation. Scores obtained were compared between each group and to social cognition reference tests, and correlated to the NeuroPsychiatric Inventory., Results: Our paradigm was suitable for assessing cognitive ToM while it lacked sensitivity for empathy assessment. Severe ToM impairment was highlighted in bvFTD while milder difficulties were observed in AD and for PD. bvFTD and AD groups produced erroneous inferences from cognitive mental states. ToM performances were linked to visual exploration strategies of the painting. Atypical visual observation was highlighted in bvFTD and AD groups causing a time shift in perspective taking of the character. Finally, we have highlighted that social cognition performances, gaze strategies and BD were correlated., Conclusion: The observation of a painting in association with eye-tracking technology can be a good support for social cognition assessment. We highlighted a link between atypical visual scanning strategies, ToM impairment and BD in these pathologies. ToM skills could be improved by training in the search for visual social cues. Therefore, this kind of remediation could have positive effects on BD.

The Elbe is one of the longest European rivers and features a large, turbid and well-mixed estuary, which runs through the inner city of Hamburg. The Elbe has been closely monitored using classical catch techniques in the past. Here we tested a COI-based eDNA approach for assessing the biodiversity within the Elbe. We sampled three stations in the Elbe, included low and high tide events, as well as two adjoining lakes to compare the recovered faunas. To analyze the data, we employed two different pipelines: the automated mBRAVE pipeline utilizing the BOLD database and one including NCBI BLAST. The number of OTUs with species or higher-level identifications were similar between both approaches with 352 OTUs and 355 OTUs for BLAST and mBRAVE, respectively, however, BLAST searches recovered another 942 unidentified metazoan OTUs. Many taxa were well represented; however, fish species were poorly represented, especially in the Elbe estuary samples. This could be a result of the universal COI primers, which also yielded high read numbers for non-metazoan OTUs, and small-bodies taxa like Rotifera, which might have been sampled together with the eDNA. Our results show a strong tidal influence on the recovered taxa. During low tide, downstream stations resembled sites further upstream, but the former showed a very different OTU composition during high tide and early tide. Such differences might be due to varying impacts of upstream-originating eDNA during tide cycles. Such factors need to be considered when routinely employing eDNA for monitoring programs., Competing Interests: The authors have declared that no competing interests exist.

There are various differences in size, behavior, and life history traits of non-Apis bee species compared with honey bees (Apis mellifera; Linnaeus, 1758). Currently, the risk assessment for bees in the international and national process of authorizing plant protection products has been based on honey bee data as a surrogate organism for non-Apis bees. To evaluate the feasibility of a semifield tunnel test for Osmia bicornis (Linnaeus, 1758) and Osmia cornuta (Latreille, 1805), a protocol was developed by the non-Apis working group of the International Commission for Plant-Pollinator Relationships, consisting of experts from authorities, academia, and industry. A total of 25 studies were performed over a 2-yr period testing a replicated control against a replicated positive control using either a dimethoate or diflubenzuron treatment. Studies were regarded to be valid, if ≥30% of released females were found to occupy the nesting units in the night/morning before the application (establishment). Thirteen studies were regarded to be valid and were analyzed further. Parameters analyzed were nest occupation, flight activity, cell production (total and per female), cocoon production (total and per female), emergence success, sex ratio, and mean weight of females and males. Dimethoate was a reliable positive control at the tested rate of 75 g a.i./ha, once >30% females had established, displaying acute effects such as reduction in flight activity, increase in adult mortality (shown by nest occupation), and reproduction ability of the females (total cell and cocoon production). On the other hand, no effects on larval and pupal development were observed. The growth regulator diflubenzuron had statistically significant effects on brood development, causing mortality of eggs and larvae at a rate of approximately 200 g a.i./ha, whereas fenoxycarb did not cause any significant effects at the tested rates of 300 and 600 g a.i./ha. In conclusion, the ring-test protocol proved to be adequate once the study comprised a well-established population of female Osmia bees, and the results improved in the second year as the laboratories increased their experience with the test organism. It is noted that the success of a study strongly depends on the experience of the experimenter, the crop quality, the quality of the cocoons, and the weather conditions. Based on these finding, recommendations for a semifield study design with Osmia spp. are proposed. Environ Toxicol Chem 2021;40:236-250. © 2020 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC., (© 2020 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.)

Background: Eye movement abnormalities (EMA) are common in multiple sclerosis (MS). However, type and severity according to the MS stage are poorly known, especially in Radiologically Isolated Syndrome (RIS) and in Clinically Isolated Syndrome (CIS). Although MRI has been included in the MS diagnostic criteria, there may be clinical-radiological dissociation., Objective: To analyze by video-oculography (VOG) prevalence of EMA in different MS phenotypes and study correlations with brain and cervical cord MRI T2 lesions location., Methods: 76 participants were prospectively recruited (12 RIS, 10 CIS, 11 relapsing-remitting-MS, 10 secondary progressive-MS, 10 primary progressive MS and 23 gender and age-matched healthy controls). We analyzed fixations, anti-saccades, horizontal and vertical reflex saccades and smooth pursuit., Results: EMA were frequent and of gradual severity from RIS to progressive forms. Internuclear ophthalmoplegia (INO) and centripetal hypermetria were strong arguments for the diagnosis of a demyelinating disorder versus a control population. Some EMA were linked to infratentorial T2 lesion location, but others like INO were not., Conclusion: This study confirm that EMA are common in all MS phenotypes, even at the earliest stages. VOG can be useful to detect demyelinating process at preclinical stage by highlighting subclinical EMA even in absence of characteristic lesions visible on MRI., (Copyright © 2020 Elsevier B.V. All rights reserved.)

For fish stock management and large-scale stocking programs, the chemical substance alizarin red S (ARS) is an important tool to mark fish permanently. Equally, for the IUCN red list species European eel (Anguilla anguilla), ARS is proven to be the most promising option for mass marking. ARS binds to calcified structures (i.e., bones and otoliths) and can be detected using a fluorescence microscope. Despite the frequent application of ARS, not only for eels but also for fish in general, until today, no study has evaluated its bioaccumulation potential. Therefore, the German Federal Risk Assessment Authority was unable to classify ARS as harmless because of a potential risk to consumers' health. Using the technique of liquid chromatography mass spectrometry, an ARS detection protocol was developed and the bioaccumulation potential of ARS in European eel muscle tissue was estimated. A detection limit of 8.9 μg kg -1 could be reached by optimizing the detection method in fish muscle tissue. In the current study, 250 eels between 6 and 57 cm of total length have been analyzed for ARS between 0 day and 3 years after the marking process. The highest concentration of ARS (6056 μg kg -1 ) was observed immediately after marking in the smallest length class. Only 1 year after the marking procedure, the ARS concentration was below detection limit. A new method for ARS detection in fish muscle tissue, followed by utilization on marked eels, was able to show that the bioaccumulation of ARS in edible fish muscle was highly unlikely.

This study presents a new chemical double-marking technique for European glass eels Anguilla anguilla by combing alizarin red S (ARS) and strontium chloride hexahydrate (Sr). Marked eels (double marked with ARS and Sr, but also single marked with ARS) were exposed in situ to brackish water (15 g l -1 artificial sea salt) for 14 days and did not exhibit increased mortalities compared with unmarked eels. Indeed, no mortality occurred in a marked group during the experiments. Moreover, an efficient mass-marking approach with low handling effort for both single ARS and double ARS-Sr techniques is described and was proven to be practicable for large-scale stocking programmes., (© 2017 The Fisheries Society of the British Isles.)

The Schlei fjord in northern Germany is the recipient water of a comprehensive eel, Anguilla anguilla (L.), stocking programme. Since 2015, stocked eels become alizarin red S marked, but to date no control mechanism is implemented in this stock enhancement measure to prevent anthropogenic spreading of diseases. Consequentially, it was possible that farmed stocking cohorts of 2015 and 2016 (in total ca. 1040 kg) were subsequently tested positive for anguillid herpesvirus 1 (AngHV 1). For this study, 100 eels [total length (TL) 24.3-72.9 cm, age ca. 1-6 years] were caught in 2016 and investigated with regard to AngHV 1 infection, parasite load (Anguillicoloides crassus) and body conditions. 68% of the eels were found to be virus positive while larger specimens were more often infected. In addition, a fitted generalized linear model (area under the curve = 0.741) demonstrated that an increase in individual TL is accompanied with an increased risk of clinically relevant virus loads. Anguillicoloides crassus turned out to be an important stressor for eels, because parasite and virus load revealed a significant positive correlation. The results of this study evidently show the urgent need of a disease containment strategy for eel stocking programmes., (© 2017 John Wiley & Sons Ltd.)

The immunomodulatory molecule Salp15 is originally described in Ixodes scapularis and has been shown to inhibit CD4 T cell activation. Many Salp15 homologs have been described from Ixodes species, and all were well conserved at C-terminal residues that seem to be essential for the function of the protein. In this study, a gene sequence was amplified from cDNA isolated from engorged female I. ricinus ticks, which was predicted to generate a protein of 12.3 kDa. The protein displayed distinct amino acid differences from previously described I. ricinus Salp15 homologs, with amino acid identity ranging between 46.6% and 93.9%. It was referred to as I. ricinus Salp15-like protein. The protein showed 48.1% sequence identity to I. scapularis Salp15. We analyzed the effect of the recombinant I. ricinus Salp15-like protein on the production of cytokines from human peripheral blood mononuclear cells stimulated with LPS. The recombinant protein exerted no effect on the production of TNF-α and IL-6, but the production of IL-10 was dose-dependently reduced. It can be concluded that I. ricinus Salp15-like protein exerts an immunomodulatory effect on the host. The inhibition of IL-10 production may possibly lead to a retardation of B cell activity., (Copyright © 2014 Elsevier GmbH. All rights reserved.)

Infections of small ruminants with Anaplasma, Theileria and Babesia species are widely distributed in the old world and are of great economic impact. In Iraq, data on disease occurrence in sheep caused by above-mentioned infectious agents are scarce. This study provides information on various haemoparasitic agents infecting sheep in the Kurdistan Region, Iraq, using molecular diagnostic tools. Altogether, 195 samples originating from three governorates in the Kurdistan Region, namely Duhok, Erbil and Sulaimaniya, were analysed. The following pathogens were identified: Anaplasma ovis (62.6%), Theileria ovis (14.35%), T. lestoquardi (7.7%), T. uilenbergi (5.6%) and Babesia ovis (1.5%). T. uilenbergi is detected for the first time in Iraq. Coinfection of sheep with different pathogens could be observed in this study, and it was found that 45 of 195 (23%) of the samples contained more than one pathogen. Even triple-positive samples were identified in 3% of the investigated animals. In conclusion, we confirm the coinfection of sheep with various haemoparasitic pathogen species in the Kurdistan Region of Iraq. Further investigations are needed to reveal the epidemiology of the diseases, the respective tick vectors, and, in the case of coinfection, pathogens' interaction and possible cross-protection., (© 2013 Blackwell Verlag GmbH.)

Anaplasma species are obligate intracellular rickettsial pathogens transmitted by ticks with an impact on human and animal health. Anaplasma ovis infects sheep and goats in many regions of the world, and it can be diagnosed by different methods like Giemsa staining, PCR or competitive ELISA. In this study, a PCR based on the gene coding for major surface protein 4 (MSP-4) was used to examine field samples collected from sheep in different countries. Altogether, 1161 blood samples from Turkey (n = 830), Iraq (n = 195), Sudan (n = 96) and Portugal (n = 40) were examined, of which 31.4%, 66.6% 41.6% and 82.5%, respectively, were positive. This indicates high prevalence of A. ovis in the countries under investigation, and it can be assumed that the situation in other areas of the world might be similar. Thus, A. ovis should be considered as an important constraint of livestock production, and further efforts are needed to better understand the epidemiology and to implement suitable control measures., (© 2013 Blackwell Verlag GmbH.)

A loop-mediated isothermal amplification (LAMP) assay was developed for the diagnosis of Theileria lestoquardi infection. The primers were designed based on the clone-5 sequence of T. lestoquardi. The specificity and sensitivity of the assay were established. Analysis of the specificity showed that the selected LAMP primers amplified the target sequence from T. lestoquardi DNA successfully, while no amplification was seen with DNA from Theileria annulata, Theileria ovis, Babesia ovis, Anaplasma ovis, or ovine genomic DNA. The specificity of the LAMP product was further confirmed by restriction digestion and sequencing. The sensitivity of the LAMP assay was analyzed in comparison to PCR resulting in a detection limit of 10 fg/μl of plasmid DNA containing the clone-5 sequence. The suitability for utilizing the LAMP assay in the field for the diagnosis of T. lestoquardi infection was tested on 100 field samples collected in Sudan and compared with results obtained by PCR. The relative specificity and sensitivity of the established LAMP assay was determined to be 92.1% and 87.5%, respectively, indicating that it may be regarded as an alternative molecular diagnostic tool to PCR which could be used for epidemiological surveys on T. lestoquardi infection.

The pathogenic protozoan parasite Theileria uilenbergi is one of the causative agents of theileriosis in small ruminants in China. The infection results in great economical losses in the northwest part of China. Efforts are underway to establish an enzyme-linked immunosorbent assay (ELISA) based on a T. uilenbergi immunodominant recombinantly expressed protein using different approaches in order to perform epidemiological studies in the area. In this study, we describe the possible use of the clone-9 protein for this purpose, which was identified as a potential immunogenic piroplasm protein by random sequencing of cDNA library clones followed by bioinformatic analyses. The clone-9 gene was partially recombinantly expressed and used for the development of an indirect ELISA for the detection of circulating antibodies in sera of T. uilenbergi-infected sheep. No cross-reactivity was observed in serum from animals infected with Theileria lestoquardi. The cut-off was calculated at 48.6% positivity using 25 serum samples from uninfected animals. A total of 101 field samples collected from an endemic area in China were used to evaluate the clone-9 ELISA for its use in the field.

Vector-borne diseases are rising in interest due to global warming, which is believed to impact on the distribution of vectors into new areas thus influencing the occurrence and epidemiology of vector-borne pathogens. Babesia canis belongs to the Piroplasmidae and there are three described subspecies, namely B. canis canis, B. canis rossi and B. canis vogeli. They are each transmitted by a different tick-species, Dermacentor reticulatus, Haemaphysalis leachi and Rhipicephalus sanguineus, respectively. There are also differences in the geographical distribution and pathogenicity to dogs of each subspecies. In this study, we aimed to establish a rapid and easy to perform DNA-based test using loop-mediated isothermal amplification to detect all three Babesia canis subspecies in one assay.

Intracellular leukoproliferative Theileria are unique as eukaryotic organisms that transform the immune cells of their ruminant host. Theileria utilize the uncontrolled proliferation for rapid multiplication and distribution into host daughter cells. The parasite distribution into the daughter cells is accompanied by a tight association with the host cell mitotic apparatus. Since the molecular basis for this interaction is largely unknown, we investigated the possible involvement of the immunodominant Theileria annulata surface protein, TaSP, in the attachment of the parasite to host cell microtubule network. Confocal microscopic analyses showed co-localization of the TaSP protein with alpha-tubulin and reciprocal immuno-co-precipitation experiments demonstrated an association of TaSP with alpha-tubulin in vivo. In addition, the partially expressed predicted extracellular domain of TaSP co-localized with the mitotic spindle of dividing cells and was co-immunoprecipitated with alpha-tubulin in transiently transfected Cos-7 cells devoid of other T. annulata expressed proteins. Pull-down studies showed that there is a direct interaction between TaSP and polymerized microtubules. Analysis of the interaction of TaSP and host microtubulin during host cell mitosis indicated that TaSP co-localizes and interacts with the spindle poles, the mitotic spindle apparatus and the mid-body. Moreover, TaSP was demonstrated to be localized to the microtubule organizing center and to physically interact with gamma-tubulin. These data support the notion that the TaSP-microtubule interaction may be playing a potential role in parasite distribution into daughter host cells and give rise to the speculation that TaSP may be involved in regulation of microtubule assembly in the host cell.

Theileriosis of small ruminants in the northwest of China is a protozoan disease that restricts the development of the livestock industry. The disease is caused by infection with Theileria uilenbergi and Theilerialuwenshuni, both of which are transmitted by ixodid Heamaphysalis ticks. The development of serological tools as a means of integrated control of the disease is an urgent and important requirement. Here we describe the identification and partial recombinant expression of a T.uilenbergi immunodominant protein (TuIP), which was identified by immunoscreening of a merozoite cDNA library. Using the recombinant TuIP (rTuIP), a novel indirect ELISA was established using 329 negative serum samples to determine the cut-off value. The internal quality control revealed satisfactory stability and repeatability of the assay. Preliminary validation using 128 positive and 48 negative reference samples demonstrated that the rTuIP ELISA is able to detect T. uilenbergi infection with high sensitivity and specificity. No cross-reactivity was found in sera from animals infected with Theileria lestoquardi, Babesia sp. China or Anaplasma ovis. Furthermore, circulating antibodies were detected in sera collected from endemic regions in China. Analyses of the antibody responses of experimentally infected animals demonstrated that tick infestation resulted in a sharply rising and stronger production of specific antibodies against TuIP while inoculation with infected blood induced an earlier production of TuIP-specific antibodies. The persistence of the TuIP-specific antibodies lasted more than 100days p.i. These data indicate the usefulness of the TuIP antigen for the development of diagnostic methods and as a potential candidate for vaccine design., (2009 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.)

Idiopathic Parkinson's disease (PD) is a neurodegenerative disorder characterized by the dysfunction of dopaminergic dependent cortico-basal ganglia loops and diagnosed on the basis of motor symptoms (tremors and/or rigidity and bradykinesia). Post-mortem studies tend to show that the destruction of dopaminergic neurons in the substantia nigra constitutes an intermediate step in a broader neurodegenerative process rather than a unique feature of Parkinson's disease, as a consistent pattern of progression would exist, originating from the medulla oblongata/pontine tegmentum. To date, neuroimaging techniques have been unable to characterize the pre-symptomatic stages of PD. However, if such a regular neurodegenerative pattern were to exist, consistent damages would be found in the brain stem, even at early stages of the disease. We recruited 23 PD patients at Hoenn and Yahr stages I to II of the disease and 18 healthy controls (HC) matched for age. T1-weighted anatomical scans were acquired (MPRAGE, 1 mm3 resolution) and analyzed using an optimized VBM protocol to detect white and grey matter volume reduction without spatial a priori. When the HC group was compared to the PD group, a single cluster exhibited statistical difference (p<0.05 corrected for false detection rate, 4287 mm3) in the brain stem, between the pons and the medulla oblongata. The present study provides in-vivo evidence that brain stem damage may be the first identifiable stage of PD neuropathology, and that the identification of this consistent damage along with other factors could help with earlier diagnosis in the future. This damage could also explain some non-motor symptoms in PD that often precede diagnosis, such as autonomic dysfunction and sleep disorders.

In this study potential molecular markers for identification of attenuation in a Theileria lestoquardi-infected cell line to be used in vaccination trials were identified. Two markers associated with attenuation in Theileria annulata vaccine strains were analyzed (metalloproteinase activity and TNF? mRNA expression). The result showed a decreased activity of MMP 9 and decreased mRNA expression of TNF? with increasing passage number. Suppression subtractive hybridization was used to identify potential new markers of attenuation. Random screening revealed nine differentially expressed genes, one from the parasite and eight from the host. Quantitative real time-PCR confirmed mRNA expression of the parasite vacuolar H+ATPase to be downregulated at higher passages.

Sheep and goats in northwest China suffer from theileriosis from infection with Theileria sp. (China), resulting in large economic losses. To investigate the immune response to infection with Theileria sp. (China), parameters of cellular and humoral immunity of experimentally infected sheep against two recombinantly expressed Theileria proteins were investigated. The in vitro proliferative response of blood mononuclear cells to a recombinant T. annulata membrane protein and a recombinant Theileria sp. (China) homologue to T. annulata surface protein, both putative membrane proteins, was significantly elevated and significant amounts of specific immunoglobulins were produced against both.

A number of Theileria annulata genes have been cloned, sequenced and expressed, including TaSP, TaD, TaSE and TamtHSP70. Several recent publications document the suitability of the recombinant TaSP protein for use in the diagnosis of tropical theileriosis. To investigate whether TaD, TaSE or TamtHSP70 elicit a humoural immune response in the T. annulata-infected host and to assess the potential of these proteins for development of diagnostics, a total of 156 field sera from Sudan and 49 negative sera from Germany were investigated in ELISA for the presence of specific antibodies against these recombinant proteins in comparison to TaSP. Antibodies against TaD and TaSE were found to be present, whereas no antibody response could be detected against the recombinant TamtHSP70. Highest titres were found to be present against the TaSP protein, with antibody titres against TaD and TaSE being in general somewhat lower. Correlation analysis showed a significant correlation of TaSP and TaSE and of TaSE and TaD antibody titres, however not between TaSP and TaD. In conclusion, the infected bovine host was shown to produce antibodies against three of the four recombinant T. annulata proteins tested, all three having been described or predicted to be parasite membrane proteins. The outstanding performance of the TaSP protein for detection of T. annulata infection in indirect ELISA was confirmed.

In previous studies, Theileria annulata surface protein (TaSP) was identified as an immunodominant antigen and successfully used to develop and validate a recombinant-protein-based ELISA for the detection of circulating antibodies in serum of T. annulata-infected animals. In this study, the same antigen was used to develop a competitive ELISA (cELISA) using a monoclonal antibody that was found to bind to TaSP. The cELISA accurately differentiated T. annulata-infected from uninfected animals and demonstrated a satisfactory performance with a calculated sensitivity and specificity of 77.4% and 100%, respectively. Thus the test proved its suitability for the diagnosis of tropical theileriosis and has application for use in serological surveys to monitor the prevalence of the disease or identify carrier animals with high specificity.

Intracellular leucoproliferative Theileria are unique as eukaryotic organisms that transform the immune cells of their ruminant host. Theileria utilize the uncontrolled proliferation for rapid multiplication and distribution into host daughter cells. The equal distribution of the schizont into the daughter cells is thought to be accomplished by a tight association with the host cell mitotic apparatus. In this study, we describe a highly conserved novel 37 kD Theileria annulata protein (TaSE). TaSE was found to be localized inside the parasite, the parasite membrane and within the host cell cytoplasm. Moreover, it co-localized at distinct points with host cell microtubules, which was especially apparent during mitosis, where co-localization was found with the centromere, the mitotic spindle and the midbody. Association of TaSE with the host cell tubulin network was corroborated by coimmunoprecipitation and transient transfection experiments. This is the first description of a theilerial protein co-localizing and potentially interacting with a host cell protein. The distribution of TaSE during mitosis makes it a protein to consider as playing a potential role for parasite distribution into daughter host cells.

Background: IgE immune responses against major allergens from Phleum pratense in low- and high-responder mouse strains were compared and the influence of alum was assessed, in order to evaluate the effect of the genetic background and adjuvants on IgE reactivity in a mouse model for P. pratense allergy., Methods: Different mouse strains and F1 offspring were sensitized with P. pratense pollen extract. Serum IgE levels, the induction of specific IgE antibodies and immediate cutaneous hypersensitivity reactions were monitored by ELISA, Western blot and a skin test, respectively., Results: All mouse strains investigated mounted an IgE response and exhibited a positive skin test to pollen extract. Differences were seen in the level of total serum IgE and in specific IgE reactivity to different major allergens of P. pratense. Notable differences were seen in IgE reactivity and immediate hypersensitivity against Phl p 1, which were only observed in SJL/j mice. The foremost influence of alum was on total IgE production levels., Conclusions: Alum is not necessary as an adjuvant to elicit IgE reactivity against the clinically relevant allergens of P. pratense, since even low-responder mouse strains mounted a hypersensitivity reaction after sensitization without the adjuvant using otherwise identical sensitization strategies. Moreover, when analyzing the allergenicity of a compound, the hypersensitivity response of different mouse strains should be considered, as implicated by the differential results obtained for IgE reactivity against Phl p 1. Lastly, a genetic component may be involved in IgE reactivity to Phl p 1., (Copyright (c) 2005 S. Karger AG, Basel)

Background: A variety of allergic reactions can be induced in mice, as measured by the induction of specific IgE. Functional read-out parameters include skin reactions and airway constriction. The aim of this study was to establish an improved quantitative assessment of the immediate cutaneous hypersensitivity reaction in a mouse model., Material/methods: Timothy grass pollen was used to sensitize BALB/c mice, which were monitored for hypersensitivity reactions by Western blot analysis of the IgE response, determination of specific immunoglobulin subclasses by ELISA, and a skin test. The immediate cutaneous hypersensitivity reaction (ICHR) was assessed by pretreating the animals with an intravenous injection of Evans blue before anesthesia and provoking a reaction by intradermal application of the allergen. Ensuing blue coloring at the injection site could be seen as an indication of mast cell degranulation and the areas of 'wheal and flare' reaction were documented for digital imaging and quantitative analysis., Results: Analysis using an antigen-specific ELISA showed that sensitization with pollen extract induced the production of specific IgE, IgG1, IgG2a and IgG2b antibodies. Western blot analyses demonstrated the production of specific IgE antibodies analogous to the IgE reactivity pattern found in allergic patients. The production of IgE was concurrent with a positive ICHR to these allergens. Quantitative analysis demonstrated that the area of ICHR increased in size in correlation to the dose of allergen applied., Conclusions: This model and the mode of skin reactivity analysis may serve as a tool to evaluate new hyposensitization and therapeutic strategies for pollen allergy.

Background: Grass pollen, such as that from timothy grass (Phleum pratense), represents a major cause of type I allergy., Objective: To characterize the IgE immune response and to identify the major allergens eliciting an IgE response in a mouse model using pollen extract of P. pratense for sensitization, in order to assess analogies to human hyperreactivity and to gain information on the allergenic potential as determined by the IgE-reactivity kinetics of defined allergens., Methods: Balb/c mice were sensitized with pollen extract or with purified natural allergens. Serum IgE levels, the induction of specific IgE antibodies and immediate hypersensitivity were monitored by ELISA, Western blot and a skin test, respectively., Results: The sensitized mice mounted a strong IgE response and showed IgE-reactivity first against Phl p 5a and 5b, then Phl p 4 and 13 and lastly against Phl p 6. No IgE response was mounted against Phl p 1. However, all purified fractions examined (Phl p 5a, 5b, 6 and 1) induced specific IgE and showed similar kinetics of IgE induction as pollen extract (first Phl p 5a and 5b, then Phl p 6). Skin test experiments demonstrated positive reactivity only in sensitized mice., Conclusion: The IgE reactivity induced by the major allergens in Balb/c mice was very similar to that found in allergic patients, with the exception of Phl p 1. The kinetics of the specific IgE response was comparable using either pollen extract or the purified major allergens, indicating that the intrinsic properties of the allergens are of importance rather than their proportionate amounts in pollen extract. This model should prove to be suitable for investigations regarding the mechanisms of induction and manifestation of timothy grass pollen allergy and for the evaluation of therapeutic strategies.

During its normal life cycle, Diplostomum spathaceum cercariae attach to and invade fish intermediate hosts. They are also known to attach to various other aquatic animals in response to water currents, touch and carbon dioxide. The purpose of this study was to identify the specific stimuli used by D. spathaceum cercariae to recognise the appropriate fish host. We characterised the host cues which stimulate them to remain on the host (enduring contact) and to penetrate the skin. Cercariae were exposed to animal skin tissues and fish skin surface mucus, their extracts and chemical modifications integrated into agar or offered via membrane filters. Enduring contact was stimulated by hydrophilic extracts Mr<3kDa, which were sensitive to oxidation of carbohydrates. The stimulating cues are probably small molecular carbohydrates, as monosaccharides stimulated enduring contacts, but amino acids, urea, electrolytes and peptides did not. Penetration was stimulated by hydrophilic macromolecules, Mr>30kDa, and by lipids. The hydrophilic stimuli were protease resistant and precipitable with Alcian blue and they were sensitive to alkaline cleavage, to digestion with lysozyme and neuraminidase as well as to oxidation of sialic acids. They were considered to be glycoproteins with O-glycosidically linked carbohydrate chains and bound sialic acids as signal structures. The lipophilic penetration stimuli were contained exclusively in the fatty acid fractions, and the stimulating characteristics of these fatty acids resembled the stimulating penetrations in other cercarial species. Diplostomum spathaceum cercariae respond to a unique profile of cues in their sequence of host-recognition phases. These cues differ from those used in other fish parasites studied to date and underline the diversity of fish recognition strategies.

Botulinum toxin produces a transient presynaptic focal block at the neuromuscular junction. Thus it induces muscle weakness which has a significant beneficial effect on hemifacial spasm. Fifty-four patients were treated and received at least 2 and at the most 22 consecutive injections over a 5-year period. Injections were repeated every 9 weeks on average. The interval between injections corresponded to the time elapsed between the last injection and the re-emergence of spasms. Nineteen patients abandoned the treatment for various reasons. The most frequent side-effect was a ptosis which was observed in 1 out of 6 injections. Ptosis is due to diffusion of the botulinum toxin over a territory larger than expected. The results obtained in 42 patients were analyzed: there was no effect in 7 patients (17%); 11 patients improved by less than 50%; 13 by 50 to 70% and 11 more than 75%.

Aphasia as the sole manifestation of focal status epilepticus in adult is rarely described. Such a case is reported here. Two separate episodes were studied: 1) prolonged mixed aphasia with continuous epileptic patterns; 2) Epileptic aphasia organized in shortly spaced temporal seizures marked by total suspension of speech, paraphasic utterances characterizing the interictal period. The clinical and EEG criteria and nosographic situation of this syndrome are reviewed. We conclude that epileptic aphasia exists if a fixed and lasting epileptic condition is created, even if a post-ictal phenomenon accounts for the symptomatology.

Clinical and biological criteria of myopathies associated with carnitine deficiency allow to distinguish a muscular and a systemic form of the condition. In this report, the results of clinical, pathological and electrophysiological data obtained from a patient with carnitine deficiency-linked myopathy are described. The patient was a 23-year-old girl who was previously known to suffer from muscle weakness when suddenly acidosis associated with a severe drop in plasma carnitine appeared. In addition there were hypermetabolic symptoms similar to those described in Luft's syndrome. Biopsy from the quadriceps femoris muscle before treatment revealed that all type I fibers were either hypotrophic or atrophic. They showed lipid overloading manifested by triglyceride droplets adjacent to the mitochondrial membrane. Furthermore, the level of soluble muscle carnitine was 83 p. 100 less than in controls and membrane linked muscle carnitine was also 73.5 p. 100 less than in controls. The patient rapidly recovered after the initiation of daily treatment with 4.40 g carnitine chlorhydrate associated with 50 g Lipogram 20. Nine months later, lipid overloading completely disappeared and the level of plasma carnitine returned to near normal whereas the level of both soluble and linked carnitine remained very low. To provide more information on the origin of the myopathy (myogenic, neurogenic or humoral) we carried out an electrophysiological investigation of cultured skeletal muscle cells from the patient and from biopsies of patients not known to be suffering from myopathy. The electrophysiological data showed that the patient myotubes were less polarized than myotubes from control patients. Furthermore, the amplitude of the action potential was smaller than the amplitude of the action potential measured in control cells. Daily addition of 50 microM carnitine chlorhydrate to the cultured myotubes induced a recovery of the action potential amplitude. Taken together these results indicate that the carnitine deficiency reported here was probably of systemic origin in addition to a myogenic component. Muscle deficiency could be either linked to an alteration in the carnitine pathway or to overconsumption of carnitine by muscle. This latter point is discussed.

Clinico-pathological data in a typical case are compared with those of 54 published reports of systemic angioendotheliomatosis. In this rare disease the lesions, characterized by clusters of abnormal cells in the small vessels, usually have multivisceral localizations, with chiefly cutaneous and encephalic manifestations and a rapidly fatal course. However cases are known with lesions restricted to the skin and a protracted if not regressive course; about half of these cases are associated with an endocarditis. In the case reported, the results of ultrastructural and histo-immunological studies (f. VIII, lysozyme, anti-protease, Ig) are in favour of the endothelial origin of the abnormal cells, a now widely accepted hypothesis.