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Background The true burden and geographical distribution of human Borna disease virus 1 (BoDV-1) encephalitis is unknown. All detected cases so far have been recorded in Bavaria, southern Germany. Case presentation A retrospective laboratory and epidemiological investigation of a 2017 case of fatal encephalitis in a farmer in Brandenburg, northeast Germany, demonstrated BoDV-1 as causative agent by polymerase chain reaction, immunohistochemistry and in situ hybridization. Next-generation sequencing showed that the virus belonged to a cluster not known to be endemic in Brandenburg. The investigation was triggered by a recent outbreak of animal Borna disease in the region. Multiple possible exposures were identified. The next-of-kin were seronegative. Conclusions The investigation highlights clinical awareness for human BoDV-1 encephalitis which should be extended to all areas endemic for animal Borna disease. All previously diagnosed human cases had occurred > 350 km further south. Further testing of shrews and livestock with Borna disease may show whether this BoDV-1 cluster is additionally endemic in the northwest of Brandenburg.

Infective endocarditis (IE) is a life-threatening disease, caused by septic vegetations and inflammatory foci on the surface of the endothelium and the valves. Due to its complex and often indecisive presentation the mortality rate is still about 30%. Most frequently bacterial microorganisms entering the bloodstream are the underlying origin of the intracardiac infection. While the disease was primarily restricted to younger patients suffering from rheumatic heart streptococci infections, new at risk categories for Staphylococcus (S.) aureus infections arose over the last years. Rising patient age, increasing drug resistance, intensive treatment conditions such as renal hemodialysis, immunosuppression and long term indwelling central venous catheters but also the application of modern cardiac device implants and valve prosthesis have led to emerging incidences of S. aureus IE in health care settings and community. The aetiologic change has impact on the pathophysiology of IE, the clinical presentation and the overall patient management. Despite intensive research on appropriate in vitro and in vivo models of IE and gained knowledge about the fundamental mechanisms in the formation of bacterial vegetations and extracardiac complications, improved understanding of relevant bacterial virulence factors and triggered host immune responses is required to help developing novel antipathogenic treatment strategies and pathogen specific diagnostic markers. In this review, we summarize and discuss the two main areas affected by the changing patient demographics and provide first, recent knowledge about the pathogenic strategies of S. aureus in the induction of IE, including available experimental models of IE used to study host-pathogen interactions and diagnostic and therapeutic targets. In a second focus we present diagnostic (imaging) regimens for patients with S. aureus IE according to current guidelines as well as treatment strategies and surgical recommendations.

Neuroleptic malignant syndrome (NMS) is a rare disorder caused by drug-induced dopamine-receptor-blockage or low dopamine concentration in the brain. It is a severe reaction to neuroleptic drugs in antipsychotic therapy. Symptoms in NMS typically consist of fever, muscle rigidity and cognitive changes; laboratory findings include elevated infectious disease markers and creatine kinase as well as signs of rhabdomyolysis. To differentiate NMS from other malignant hyperthermia syndromes identifying the offending drug and clinical history are essential. Therapy in NMS includes withdrawal of the causative medication and intensive care treatment possibly with administration of dantrolene.In this case report, we describe the clinical course of a 48 year old man who developed typical clinical symptoms and laboratory parameters of malignant hyperthermia syndrome after injection of haloperidol.

A novel H5N8 highly pathogenic avian influenza virus (HPAIV) was detected in a greater white-fronted goose in January 2020 in Brandenburg, Germany, and, in February 2020, in domestic chickens belonging to a smallholding in Baden-Wuerttemberg, Germany. Full-genome sequencing was conducted on the MinION platform, enabling further phylogenetic analyses. The virus of clade 2.3.4.4b holds six segments from a Eurasian/Asian/African HPAIV H5N8 reassortant and two segments from low pathogenic avian influenza H3N8 subtype viruses recently detected in wild birds in Central Russia. These new entries continue to show the reassortment potential of the clade 2.3.4.4 H5Nx viruses, underlining the necessity for full-genome sequencing and continuous surveillance.

Usutu virus (USUV) is an arbovirus within the genus flavivirus, which was first introduced to Southern Europe approximately twenty years ago causing epizootics among wild and captive birds. In Germany USUV was initially discovered in wild birds, mainly Common blackbirds (Turdus merula), in the Upper Rhine valley in southwest of the country in 2011 and has not spread much northwards since. Phylogenetic analyses revealed that the still ongoing USUV epidemic is caused by two different USUV strains, USUV-Germany belonging to the USUV Europe 3 lineage and USUV-Bonn belonging to the USUV Africa 3 lineage. The two strains were introduced independently. In August 2015 a new USUV strain, named USUV-Berlin, was isolated in Vero cells from two carcasses of juvenile Great grey owls (Strix nebulosa) kept in the Zoological Garden Berlin, which had suffered from a hyperacute fatal systemic infection. Both owls carried high USUV genome loads. Full-length USUV genomes sequences were determined and phylogenetic analysis demonstrated a close relationship with a Spanish mosquito-derived sequence from 2006. Immunohistochemical antigen detection in organ samples of the owls showed the typical USUV infection patterns. According to the phylogenetic analysis, USUV-Berlin belongs to the Africa 2 lineage, and can thus be distinguished from the other strains circulating in Germany. Repeated findings of different USUV strains suggest more frequent introductions into Central Europe and a higher mobility of this virus than assumed to date.

SUMMARYIn Germany tularemia is a re-emerging zoonotic disease. Therefore, we investigated wild animals and environmental water samples for the presence and phylogenetic diversity of Francisella tularensis in the poorly studied Berlin/Brandenburg region. The phylogenomic analysis of three isolates from wild animals revealed three new subclades within the phylogenetic tree of F. tularensis [B.71 from a raccoon dog (Nyctereutes procyonoides); B.74 from a red fox (Vulpes vulpes), and B.75 from a Eurasian beaver (Castor fiber albicus)]. The results from histological, PCR, and genomic investigations on the dead beaver showed that the animal suffered from a systemic infection. Indications were found that the bacteria were released from the beaver carcass into the surrounding environment. We demonstrated unexpectedly high and novel phylogenetic diversity of F. tularensis in Germany and the fact that the bacteria persist in the environment for at least one climatic season. These findings support a broader host species diversity than previously known regarding Germany. Our data further support the assumption derived from previous serological studies of an underestimated frequency of occurrence of the pathogen in the environment and in wild animals. F. tularensis was isolated from animal species not previously reported as natural hosts in Germany.

Encephalitis can be caused by several infectious agents, including bacteria, fungi, parasites and viruses. In many cases, the causative agent cannot be identified, because the pathogens are unknown or detection methods are not routinely available. In our case, a 15-month-old cow developed central nervous disorders and died within 6 days after the onset of clinical signs. The histopathology revealed an acute encephalitis, predominantly in the brain stem, and a ganglionitis of the trigeminal ganglion with massive neuronal necroses in both the brain and the ganglion. However, a relevant panel of bacterial and viral infections of cattle could be routinely excluded. Therefore, a brain sample from the cow was analysed using a metagenomics approach with next-generation sequencing. A novel bovine astrovirus (BoAstV-BH89/14) could be identified using the analysis pipeline RIEMS, and the finding could be confirmed with a specific BoAstV RT-qPCR. The genome of the bovine astrovirus (BoAstV), belonging to the family Astroviridae in the genus Mamastrovirus, has a length of 6478 bp. Sequence identities between 71% to a sheep astrovirus and 69% to two recently described bovine astroviruses from the USA and Switzerland were ascertained. The latter were also connected to encephalitis cases in cattle. Like these, the new virus described here was detected in different brain sections using the specific BoAstV RT-qPCR and fluorescent in situ hybridization. In conclusion, while astroviruses so far were mainly found in relation to gastroenteritis in animals and humans, recently detected astrovirus infections were also related to encephalitis.

The raccoon (Procyon lotor) has successfully invaded central Europe, despite the population apparently having been founded by a small number of individuals in two distinct populations in Germany. The ecological success of the invasion has been explained by raccoons being an adaptable, truly omnivorous species. However, the German raccoon population might have a larger number of founders and be more genetically diverse than assumed, as accidental or deliberate releases of household pets or individuals from zoos are relatively common. In the present study, we sequenced a 550-base-pair long fragment of the mitochondrial control region in 193 raccoons from Germany and neighbouring countries. We only identified six different haplotypes; of which, five were limited to Germany. Our results support the notions that the population was founded by a small number of females and that the German raccoons originate from two separate release events in central and eastern Germany. Additionally, however, we provide evidence for the presence of a distinct population in Saxony, eastern Germany. Further studies using different genetic markers are necessary to gain additional information on genetic diversity and population genetic structure.

A 5-years-old moose (Alces alces) cow kept in a zoo in the German Federal State of Brandenburg aborted a female foetus of 44cm crown rump length (CRL). Pathohistological analysis revealed several Neospora (N.) caninum infected cells and cysts, as well as multifocal gliosis, necrosis, haemorrhages, dystrophic mineralisation and haemosiderosis in the brain, predominantly in cerebrum and brainstem. In addition, mild lymphocytic meningitis was present. Together with the fresh foetus, a mummified foetus of 16cm CRL was expelled. Neither focal necrosis, nor inflammation was detected in the brain of the mummified foetus. By two polymerase chain reactions (PCR) targeting the pNc5 gene of N. caninum (i.e. an end point PCR and a real-time PCR), by two serological methods (immunofluorescence test and immunoblot), by histological and immunohistochemical analyses, transplacental N. caninum infection was confirmed in the fresh foetus and interpreted as possible cause of abortion. Infection with other agents causing abortion including Bovine Herpesvirus 1 (BHV1), Bluetongue Virus (BTV), Bovine Virus Diarrhoea Virus (BVDV), Brucella spp., Chlamydia spp., Coxiella burnetii and Toxoplasma gondii were excluded. Our findings show that control measures may be necessary to protect captive moose against accidental N. caninum infection. Further studies are needed to explore the importance of neosporosis in wild and captive moose.

A novel avian alphaherpesvirus, preliminarily designated sphenicid alphaherpesvirus 1 (SpAHV-1), has been independently isolated from juvenile Humboldt and African penguins (Spheniscus humboldti and Spheniscus demersus) kept in German zoos suffering from diphtheroid oropharyngitis/laryngotracheitis and necrotizing enteritis (collectively designated as penguin-diphtheria-like disease). High-throughput sequencing was used to determine the complete genome sequences of the first two SpAHV-1 isolates. SpAHV-1 comprises a class D genome with a length of about 164 kbp, a G+C content of 45.6 mol% and encodes 86 predicted ORFs. Taxonomic association of SpAHV-1 to the genus Mardivirus was supported by gene content clustering and phylogenetic analysis of herpesvirus core genes. The presented results imply that SpAHV-1 could be the primary causative agent of penguin-diphtheria-like fatal diseases in banded penguins. These results may serve as a basis for the development of diagnostic tools in order to investigate similar cases of penguin diphtheria in wild and captive penguins.

Hypoalbuminemia predicts disability and mortality in patients with various illnesses and in the elderly. The association between serum albumin concentration at the time of listing for lung transplantation and the rate of death after lung transplantation is unknown. We examined 6808 adults who underwent lung transplantation in the United States between 2000 and 2008. We used Cox proportional hazard models and generalized additive models to examine multivariable-adjusted associations between serum albumin and the rate of death after transplantation. The median follow-up time was 2.7 years. Those with severe (0.5–2.9 g/dL) and mild hypoalbuminemia (3.0–3.6 g/dL) had posttransplant adjusted mortality rate ratios of 1.35 (95% CI: 1.12–1.62) and 1.15 (95% CI: 1.04–1.27), respectively. For each 0.5 g/dL decrease in serum albumin concentration the 1-year and overall mortality rate ratios were 1.48 (95% CI: 1.21–1.81) and 1.26 (95% CI: 1.11–1.43), respectively. The association between hypoalbuminemia and posttransplant mortality was strongest in recipients with cystic fibrosis and interstitial lung disease. Hypoalbuminemia is an independent risk factor for death after lung transplantation.

Nutrient supply in organic pig production is characterized by limited amino acids (AA) availability due to the preferable use of home-grown feedstuffs and restrictions on purchased feedstuffs. This can cause large variations in the quality of diets, carcasses, and pork. The objective of two feeding trials was to assess the interactions between feeding regimes, genotype, and birth weight on carcass and pork quality. A control regime was compared with two feeding regimes that were partly (only in the finishing phase (FIN)) or both in the growing and finishing phase (GRO + FIN) restricted to home-grown feedstuffs, thus differing in AA supply. Using an isocaloric ration, individually housed pigs differing in genotype (Experiment 1) or birth weight (Experiment 2) were allocated to the three feeding regimes.The highest daily live weight gain, the best feed conversion, and the highest values for performance traits and meat composition were achieved by Du × DL pigs, while the highest carcass yield was achieved by Pi × (DL × DE) pigs. In Experiment 1, performance traits were significantly higher in the control feeding regime than in the GRO + FIN treatment, with the feeding regime FIN being intermediate. Lean meat percentage was significantly lower in the GRO + FIN feeding regime than in the control while the fat area was not influenced by feeding regime. Intramuscular fat content was higher under the GRO + FIN feeding regime without AA supplementation than in the control. In Experiment 2, birth weight showed no significant effect on carcass yield, carcass traits and meat composition, but affected growth rate. Performance traits were highest in the control, while meat composition was best in the GRO + FIN treatment, confirming results of Experiment 1. Exclusion of AA supplementation in the feeding regime reduced growth but increased intramuscular fat content (IMF). The feeding regime was the main source of variation for intramuscular fat content in the longissimus muscle . Organic pig production can yield high quality pork, but information on feed, feed intake, and pig characteristics is important to steer the production process.

In this study we identified and characterized a major latex lectin - designated as EtLLH - with antimicrobial activity from the succulent African milk tree Euphorbia trigona. The lectin is highly concentrated in the latex of E. trigona and appears to be composed of at least two subunits with a molecular mass of 32 kDa. EtLLH shares significant similarities to known plant lectins - ricin from Ricinus communis and agglutinin from Viscum album coloratum - which specifically bind D-galactose and N-acetyl-D-glucosamine, the major building blocks of many fungal cell walls. Antimicrobial activity assays revealed an impact of EtLLH on three phytopathogenic filamentous ascomycetes. The germination of the conidiospores and the hyphal growth of Aspergillus niger and Fusarium graminearum were severely inhibited by EtLLH already at concentrations below 0.1 mg cm-3, while the effect on germination of the melanized conidiospores of Botrytis cinerea was less significant.

The aim of the study was to investigate antimicrobial susceptibilities of bovine Helcococcus ovis isolates and to detect genes encoding for H. ovis erythromycin and tetracycline resistance. Twenty-nine isolates were included and the minimal inhibitory concentrations (MICs) of seven antimicrobials were determined using test conditions as given in an approved CLSI guideline for the pyridoxal-dependent Abiotrophia spp. and Granulicatella spp. Furthermore, the macrolide resistance phenotype was examined by the erythromycin–clindamycin double-disk test (D-zone test). Erythromycin MICs of ≥8 μg/ml were found in three (10%) isolates which also presented the macrolide, lincosamide, and streptogramin B (MLS B ) resistance phenotype, either constitutive or inducible. The erm (B) gene was detected in one of these isolates. Increased tetracycline MICs (≥8 μg/ml) were obtained for 24 (83%) isolates, mostly associated with the tet (M) gene alone ( n = 21) or both the tet (L) and tet (M) genes ( n = 2). The MICs determined for penicillin, ampicillin, amoxicillin-clavulanic acid, and cephalothin did not indicate resistance to these antimicrobials. The study suggests that resistance to MLS B antimicrobials and tetracycline is frequent in H. ovis . Moreover, this is the first report about occurrence of the resistance genes erm (B), tet (L), and tet (M) in the Helcococcus genus.

Background Congenital malformations involving the Mullerian ducts are observed in around 5% of infertile women. Complete aplasia of the uterus, cervix, and upper vagina, also termed Mullerian aplasia or Mayer–Rokitansky–Kuster–Hauser (MRKH) syndrome, occurs with an incidence of around 1 in 4500 female births, and occurs in both isolated and syndromic forms. Previous reports have suggested that a proportion of cases, especially syndromic cases, are caused by variation in copy number at different genomic loci. Methods In order to obtain an overview of the contribution of copy number variation to both isolated and syndromic forms of Mullerian aplasia, copy number assays were performed in a series of 63 cases, of which 25 were syndromic and 38 isolated. Results A high incidence (9/63, 14%) of recurrent copy number variants in this cohort is reported here. These comprised four cases of microdeletion at 16p11.2, an autism susceptibility locus not previously associated with Mullerian aplasia, four cases of microdeletion at 17q12, and one case of a distal 22q11.2 microdeletion. Microdeletions at 16p11.2 and 17q12 were found in 4/38 (10.5%) cases with isolated Mullerian aplasia, and at 16p11.2, 17q12 and 22q11.2 (distal) in 5/25 cases (20%) with syndromic Mullerian aplasia. Conclusion The finding of microdeletion at 16p11.2 in 2/38 (5%) of isolated and 2/25 (8%) of syndromic cases suggests a significant contribution of this copy number variant alone to the pathogenesis of Mullerian aplasia. Overall, the high incidence of recurrent copy number variants in all forms of Mullerian aplasia has implications for the understanding of the aetiopathogenesis of the condition, and for genetic counselling in families affected by it.

Summary 1. Visually foraging fish typically exclude large zooplankton from clear-water lakes and reservoirs. Do fish have the same effect in turbid waters, or does turbidity provide a refuge from visual predation? 2. To test the hypothesis that fish exclude large zooplankton species from turbid sites, I searched for populations of medium or large Daphnia species in turbid, fish-containing reservoirs of south-central Oklahoma and north-central Texas, U.S.A., and surveyed the literature for accounts of Daphnia species in turbid habitats worldwide. 3. Only small Daphnia species and the exuberantly spined Daphnia lumholtzi were detected in the turbid reservoirs. The Daphnia species in the reservoirs are smaller than other Daphnia species that occur in the area but were not detected. An extensive survey of the literature suggests that large Daphnia may be found in the lakes of extreme turbidity [Secchi disk depth (SD) SD > 0.2 m) unless some additional factor reduces the influence of fish predation in such sites. 4. The field samples from Texas and Oklahoma together with the literature review suggest that the effect of visually foraging planktivorous fish on the size structure of turbid-water zooplankton communities may often be as strong or even stronger than the effect of fish on clear-water zooplankton communities.

Zusammenfassung In drei Fallen von Kryptorchismus und einem Fall von Sertolizellsyndrom wurden haufig Leydigzellen in der verdickten Lamina propria der Tubuli seminiferi gefunden. In normalem Gewebe dagegen wurden Leydigzellen in den peripheren Schichten des peritubularen Gewebes nur gelegentlich als spindelformige Zellvarianten angetroffen. In alien Fallen konnten die Leydigzellen eindeutig im Elektronenmikroskop identifiziert werden: sie enthalten u.a. ein gut entwickeltes glattes endoplasmatisches Retikulum, prakristalline tubulare Strukturen und Reinke Kristalle. Die Entwicklung der Leydigzellen aus Myofibroblasten in der Lamina propria wird diskutiert.

The initial isolation of Helcococcus ovis from a valvular thrombus prompted us to investigate the prevalence of this bacterium in bovine valvular endocarditis. Specimens from 55 affected hearts were examined by culture using Columbia blood agar and cross streaking the inoculated plate with a Staphylococcus aureus strain. As confirmed by 16S rRNA gene sequencing, H. ovis was isolated with an unexpectedly high frequency of 33%, predominantly as heavy growth and pure culture. The majority of H. ovis isolates showed distinct satellitism around S. aureus and pyridoxal dependency, resembling “nutritionally variant streptococci” (now assigned to the genera Abiotrophia and Granulicatella ). Using the API rapid ID 32 Strep, API ZYM, and Rosco Diatabs systems, incongruent results were obtained for alkaline phosphatase, β-galactosidase, β-glucuronidase, and leucine aminopeptidase activities. Based on the satellitism/pyridoxal dependency; hemolysis on blood agar; the API rapid ID 32 Strep results for arginine dihydrolase, α-galactosidase, β-galactosidase, β-glucuronidase, and pyroglutamic acid arylamidase activities; hippurate hydrolysis; and acidification of sucrose, a scheme for the identification of H. ovis and its differentiation from other members of the Helcococcus genus and the pyridoxal-dependent species Abiotrophia defectiva, Granulicatella adiacens , and Granulicatella elegans is proposed. By establishing specific fluorescence in situ hybridization, large H. ovis aggregates were specifically detected within the fibrinous exudate of the valvular thrombi. Our results demonstrate for the first time that H. ovis represents an emerging pathogen in bovine valvular endocarditis that is frequently isolated if appropriate culture conditions are used.

SUMMARY 1. We measured the abundance and eggs per female of four Daphnia species in turbid and relatively clear regions of Lake Texoma (Oklahoma-Texas, U.S.A.) on 12 dates over the course of 5 years. 2. Two species, Daphnia lumholtzi and Daphnia parvula, occurred and reproduced in turbid locations, but two other species, Daphnia mendotae and Daphnia pulicaria, occurred almost exclusively in relatively clear conditions. 3. To test the hypothesis that interference with foraging excludes clear-water Daphnia species from turbid locations, we incubated adult D. mendotae at both a clear and a turbid site. In three successive experiments D. mendotae individuals incubated at the turbid site carried as many or more eggs than individuals incubated at the clear site.

The polymorphic barley (Hordeum vulgare) Mla locus harbors allelic race-specific resistance (R) genes to the powdery mildew fungus Blumeria graminis f sp hordei. The highly sequence-related MLA proteins contain an N-terminal coiled-coil structure, a central nucleotide binding (NB) site, a Leu-rich repeat (LRR) region, and a C-terminal non-LRR region. Using transgenic barley lines expressing epitope-tagged MLA1 and MLA6 derivatives driven by native regulatory sequences, we show a reversible and salt concentration-dependent distribution of the intracellular MLA proteins in soluble and membrane-associated pools. A posttranscriptional process directs fourfold greater accumulation of MLA1 over MLA6. Unexpectedly, in rar1 mutant plants that are compromised for MLA6 but not MLA1 resistance, the steady state level of both MLA isoforms is reduced. Furthermore, differential steady state levels of MLA1/MLA6 hybrid proteins correlate with their requirement for RAR1; the RAR1-independent hybrid protein accumulates to higher levels and the RAR1-dependent one to lower levels. Interestingly, yeast two-hybrid studies reveal that the LRR domains of RAR1-independent but not RAR1-dependent MLA isoforms interact with SGT1, a RAR1 interacting protein required for the function of many NB-LRR type R proteins. Our findings implicate the existence of a conserved mechanism to reach minimal NB-LRR R protein thresholds that are needed to trigger effective resistance responses.

Background and Purpose: Enteral applied bovine colostrum can significantly reduce endotoxin concentration in plasma. Since colostrum is a mixture of biological active ingredients 3 possible substances which are able to influence the endotoxin elimination were concentrated in 3 different colostrum products. Immunoglobulin-, lactoferrin- and casein-enriched colostra and lactoferrin alone were orally administered to endotoxinaemic rats. Methods: Endotoxinaemia was induced to rats by enteral application of 1010E. coli together with 40 mg Nebacetin. Control animals received albumin. From all rats plasma samples were taken over the time of 5 h and endotoxin concentration determined with limulus lysate and chromogenic substrate. Results: Whereas in control animals as well as in animals treated with casein-enriched colostrum a marked increase of endotoxin values to over 130 EU/dl could be observed after 5 h, the oral application of gammaglobulin-enriched and especially lactoferrin-enriched colostrum decreased endotoxin values by more than 50%. The most effective endotoxin elimination was seen with lactoferrin alone. Conclusions: From this results it can be concluded that not only gammaglobulin but especially lactoferrin seems to be responsible for the elimination of endotoxin with regard to enterally applied colostrum preparations.

Ethnopharmacological relevance Dandelion ( Taraxacum spec) is a wild plant that has been used for centuries as a traditional medicine in the relief and treatment of several diseases. This use is due to the presence of sesquiterpenes, saponins, phenolic compounds, flavonoids, and sugars, among others, found in the organs of the plant. Aim of the study The aim of this work is to provide a current review of developments and trends in research on the Taraxacum genus, with a focus on traditional uses and pharmacological properties. This should shed light on the potential of this plant as an attractive commercial herbal medicine. Materials and methods Documents were collected, analyzed, and classified for information regarding medical, agronomic, genetic, and biological aspects of the Taraxacum species. This process was based on a thorough search of documents indexed by scientific search engines. Results Two important periods of research on Taraxacum have been identified: the first, between 1930 and 1950; and the second, from 1990 to today. During the former, agricultural and genetics research on this plant were, due to the shortage of natural rubber, the focus. In contrast, the main drive in Taraxacum research is now the recovery of bioactives and/or applications in medicine. Pharmacology is the main area in which these plants have been tested, thanks in part to its widely known traditional uses; however, there is less than enthusiastic interest in further human clinical trials. In other areas, Taraxacum sports an enormous list of compounds of industrial interest; and while it is true that only a small amount of these compounds is immediately available in Taraxacum organs and makes it relatively commercially unattractive, only scarce efforts have been made to improve yields. Compounding this issue, most studies of its growth and cultivation have been focused mainly on controlling it as a weed detrimental to certain industrial crops. To wit, in spite of all the research carried out, less than 1% of all the species identified so far (>2500) have been studied (including Taraxacum officinale , Taraxacum coreanum , Taraxacum mongolicum and Taraxacum platycarpum ). This is a indication of the little knowledge that we have about this genus so far. Biotechnology (involving genetics, agriculture, and biology) is the most powerful means by which to take advantage of all the medicinal potential of Taraxacum . Great strides have been made in identifying metabolic pathways for synthesizing terpenes, one of the most important compound families in clinical applications. In order to improve yield and performance of the plant in the field, greenhouse cultivation is another aspect taken into account, deriving an increase in recovery of bioactives from Taraxacum organs. Even while considering that only a few species have been studied, their different biochemical and cultivation profiles indicate huge potential for qualitative improvements in composition through genetic engineering, thus directly impacting pharmacological properties. Conclusions Taraxacum is has been traditionally considered a natural remedy, well-inserted into popular knowledge, but with low commercial applicability. Only once the recovery of pure and highly reactive compounds can be pursued at (a qualitatively and quantitatively attractive) economical scale, human clinical trials would be of interest in order to prove their efficacy and safety, positioning Taraxacum as an important commercial source of natural drugs.

A new progressive tremor disorder called Campus syndrome (CPS) was observed among the progeny of a normal boar of the Pietrain breed in Germany. Extensive backcross experiments indicate that CPS is inherited as an autosomal dominant trait, and the founder boar, Campus, is believed to be a gonadal mosaic. A linkage analysis of 57 animals mapped the CPS gene to a region on porcine chromosome 7 flanked by the markers SW1418 and SW352, which is homologous to a part of human chromosome (HSA) 14. Human dominant distal myopathy type 1 (MPD1) has been mapped to the homologous region of HSA14. As the myopathological findings in MDP1 show striking similarities to CPS, this porcine disorder may serve as an animal model for MPD1.

To investigate the correlation between the neuropathologic findings after laser-induced interstitial thermotherapy (LITT) and MRI temperature maps during laser irradiation, a total of six pig brains from cadavers were treated with a Nd:YAG laser (lambda = 1,064 nm, 3.8 W) for 15 minutes. For MRI monitoring, we used a phase-sensitive two-dimensional (2D) fast low-angle shot (FLASH) sequence on a 1.5-T Magnetom SP. Temperature maps were acquired every minute (accuracy

The capacity of several coat protein (CP) mutants of a German isolate of barley yellow mosaic bymovirus (BaYMV) to bind of nucleic acids was studied in vitro. Recombinant CP, produced by overexpression in Escherichia coli, was purified from inclusion bodies and subsequently renatured. Binding to single-stranded (ss) RNA and ssDNA oligonucleotides was found to be cooperative and sequence non-specific. By deletion mutagenesis, several truncated CP derivatives were created and their nucleic acid-binding capacity was investigated in order to define a protein domain responsible for RNA- and DNA-binding. The nucleic acid-binding domain consists of a core which was located to an internal 23 amino acid peptide (aa 125-147) and an adjacent domain (aa 148-184) which stimulates binding.

It has been proposed that cases of diaptomid dominance in oligotrophic lakes are due to lower food requirements for Diaptomus species than Daphnia species. We test this hypothesis by competing Daphnia pulicaria and Diaptomus oregonensis populations in continuous and semicontinuous cultures. The threshold food concentrations for population growth of D. pulicaria and D. oregonensis were statistically indistinguishable. The thresholds of both species were within the range of concentrations characteristic of ultraoligotrophic lakes. The dominant species varied across experiments in a pattern consistent with an effect of temporal variation in food concentration. Neither our data nor previously published studies provide support for the hypothesis that cases of Diaptomus dominance in oligotrophic lakes are due to a simple difference in threshold food concentrations

SUMMARYA total of 2475 animals from Germany, both captive and wild, were tested for antibodies againstFrancisella tularensisto obtain more knowledge about the presence of this pathogen in Germany. An indirect and a competitive ELISA served as screening methods, positive and inconclusive samples were confirmed by Western blot. Of the zoo animals sampled between 1992 and 2007 (n = 1122), three (0·3%) were seropositive. The seroconversion of a hippopotamus in Berlin Zoo was documented. From 1353 serum samples of wild foxes (Vulpes vulpes), raccoon dogs (Nyctereutes procyonoides) and wild boars (Sus scrofa), collected between 2005 and 2009 in the federal state of Brandenburg (surrounding Berlin), a total of 101 (7·5%) tested positive for antibodies toF. tularensislipopolysaccharide. Our results indicate a higher seroprevalence ofF.tularensisin wildlife in eastern Germany than commonly assumed. Furthermore, we found foxes and raccoon dogs to be biological indicators for tularaemia.

We demonstrate how research on innovative agroforestry systems, incor- porating new species and management approaches, can benefit from preliminary financial appraisals that include both economic and ecological variables in quantitative cost-benefit models. Model results provide vital information for planning further research, and thus increase the likelihood of finding practical solutions to urgent resource management prob- lems within reasonable funding and time constraints. As a case study, we evaluate the potential of enrichment plantings of three native fruit-producing genera in selectively logged rain forests of Kalimantan, Indonesia. The models use preliminary estimates of 8 ecological parameters (e.g., survival, growth rate, fruit production) and 11 economic parameters (e.g., planting and harvesting costs, market prices) to predict net present value (NPV) over a variety of project durations and discount rates. Sensitivity analyses examine the effect of changes in input parameters. The genera were chosen (1) for ecological complementarity in the use of light resources and (2) to produce fruit at times when commonly harvested species are unavailable in markets. Dialium spp. are canopy trees that produce timber and a relatively nonperishable fruit; Willughbeia spp. are fast-growing lianas that produce fruit; Garcinia spp. are understory fruit trees. Sensitivity analyses indicate that all three genera have potential for substantial positive NPV, and that NPV projections depend heavily upon fruit price, fruit production, age of first reproduction, juvenile survivorship, and harvesting and marketing costs. In addition, these analyses demonstrate that preliminary, combined ecological-economic analysis can guide the selection of species and research strategies for experimental trials.

Background According to the classification of polycystic ovary syndrome (PCOS) published by the Androgen Excess Society (AES), 10 different phenotypes of the condition are possible. The question remains of whether using these phenotypes might enable us to identify women with impaired insulin resistance or an impaired lipid profile among patients with PCOS. Materials and methods A prospective cohort analysis was performed of 313 women with diagnosed PCOS and 80 control individuals. The screening panel included a physical examination, weight and height measurement, ultrasonography of the ovaries, and hormone, glucose, lipid, and insulin resistance measurements. Results There were no statistically significant differences in insulin resistance parameters between the different phenotypes. There were no statistically significant differences in body mass index (BMI) in any of the groups, but BMI showed the best correlation with insulin resistance in all women with PCOS and controls. Sex hormone-binding globulin (SHBG) was inversely correlated with insulin resistance in women with PCOS and controls. High-density lipoprotein (HDL) was negatively correlated with insulin resistance, and free testosterone was positively correlated with it, only in women with PCOS. Conclusions Using the different phenotypes described in the AES classification shows no advantages for identifying women with aggravated insulin resistance or impaired lipid profile among patients with PCOS.

SUMMARY 1 This study compares small-scale (i.e. neighbourhood) associations among distributions of phytoplankton, zooplankton, and zooplankton grazing rates using a combination of correlations and analyses of spatial patchiness. 2 All zooplankton and phytoplankton populations had patchy distributions. On average, individuals experienced members of their same taxon at a density that was 42% greater than the mean sample density. 3 The various zooplankton taxa experienced different average neighbourhoods, even within the same stratum of the lake. For example, Daphnia galeata mendotae (Birge) experienced phytoplankton concentrations that were nearly 50% greater than those experienced by Daphnia pulicaria (Forbes) at the same depth. 4 The distributions of the various phytoplankton taxa were positively correlated with each other, but the distributions of zooplankton were negatively correlated, or more often, unrelated to each other. 5 Phytoplankton abundance was negatively correlated with zooplankton grazing rates. 6 We hypothesize that while phytoplankton distributions are driven by common external factors, the factors that determine horizontal distributions of zooplankton are species specific. In addition, zooplankton grazing appears at least partially responsible for the generation of patchiness in the distribution of phytoplankton.

Summary A “resistance breaking” isolate of barley yellow mosaic virus-2 (BaYMV-2) was cloned as a cDNA and characterised. Restriction mapping and comparison with a German and a Japanese isolate of BaYMV (BaYMV-G and BaYMV-J) revealed a high level of restriction site conservation for RNAl and the majority of RNA2. However, in a box of approximately 600 nucleotides located on RNA2, striking differences in the restriction pattern could be identified. The nucleotide sequence of this box, as well as of the 3‘-terminal region of RNA1 including the coat protein coding region, and the deduced amino acid sequences were analysed. Identity at the amino acid level was between 99.3% and 92.3% in comparison with the corresponding sequences of BaYMV-G and BaYMV-J, suggesting that BaYMV-2 is closely related to BaYMV. Consequently, the classification of BaYMV-2 as a “resistance breaking” strain of BaYMV is justified.

The morphology and molecular composition of intercellular adherens junctions have most frequently been described in epithelial cells and the fascia adhaerens of the intercalated disc. A group of cytoplasmic molecules is known to be associated with adherens junctions. The intercellular bond is mediated by cadherins which bridge the cells by homophilic binding. Recently, endothelial cells have also been shown to form intercellular junctions of the adherens-type. However, they are morphologically less distinct and little is known about their molecular components. In this study we report the localization of some adherens junction components in intact microvessels of the blood-brain barrier in the rat. We used antibodies raised against alpha-actinin, vinculin, zyxin, cadherin (antipan-cadherin antibody) and A-CAM (N-cadherin) in immunohistochemical experiments at light and electron microscopical levels. Microvessel walls reacted positively for all antigens throughout postnatal development. All antigens were localised, though not necessarily exclusively, to interendothelial junctions. At the ultrastructural level, pan-cadherin reactivity was present throughout the entire length of the cleft. These results could mean that in blood-brain barrier endothelial cells the complex tight junction is embedded in an adherens junction which occupies the entire length of the cleft.

The location and structure of sensory nerve endings was examined in the mucosa and in the gingiva propria adjacent to the third premolar of Monodelphis domestica by light and electron microscopy. The mucous membrane of the gingiva propria is covered with a stratified keratinized squamous epithelium. The mucous membrane of the adjacent inner aspect of the lip and of the vestibulum oris were covered with a nonkeratinized squamous epithelium. Free nerve endings, Merkel nerve endings and lamellated corpuscles were found in all the examined areas. Free nerve endings were located in the connective tissue papillae between the epithelial cones, in the basal layer of the epithelium and directly under the epithelium of the cones. They were innervated by myelinated A-delta- and nonmyelinated C-fibers. The basal layer of the first three to four epithelial cones of the gingiva propria contained single or groups of Merkel nerve endings. The epithelial cones of the oral mucous membrane contained five times more Merkel nerve endings than those of the vestibular mucous membrane. Lamellated corpuscles were mainly found in the plica sublingualis. In the connective tissue below the epithelial cones of the mucous membranes they usually occur with a perineural capsule, in the connective tissue papillae between the cones they occur without a perineural capsule. In the latter position they resemble the Meissner corpuscles of glabrous skin. The density of receptors in the oral mucous membrane of the lower jaw of M. domestica implies that this richly innervated mechanoreceptive area acts as an antagonist of the tongue and provides important information during mastication.

Recycling of 46,000 M(r) mannose 6-phosphate receptor (MPR 46) was investigated by microinjection of Fab fragments against small epitopes within the cytoplasmic domain of the receptor. Fab fragments against the peptide 43-47 (Ala-Tyr-Arg-Gly-Val) efficiently blocked return of MPR 46 to the TGN. Antibody-induced redistribution resulted in accumulation of MPR 46 within an endosomal compartment, from which it recycled to the plasma membrane. Rab5 and rab7, markers for early and late endosomes, respectively, were not detectable in the compartment of redistributed MPR 46, suggesting that it represents a specialized endosomal subcompartment. The bulk of redistributed MPR 46 did not colocalize with endocytosed fluid-phase marker, suggesting that it accumulates at a site where MPR 46 has been segregated from endocytosed material, which is destined for transport to lysosomes. Peptide 43-47 contains a tyrosine (residue 44) which has been shown earlier to be part of an internalization signal for MPR 46 (Johnson, K. F., W. Chan, and S. Kornfeld. 1990. Proc. Natl. Acad. Sci. USA. 87:10010-10014). The role of tyrosine residue 44 as part of a putative multifunctional sorting signal is discussed.

Water and hydrophilic solutes cross the endothelium of continuous capillaries via the paracellular cleft and possibly other routes. This pathway shows a selectivity to molecule size and charge. However, it is not yet known which systems confer this selectivity. Isolated rat hearts were perfusion-fixed through the coronary circulation, stained with lanthanum or tannic acid, and further processed for transmission electron microscopy. Thin sections viewed at × 160,000 magnification revealed regularly spaced, cleft-spanning structures in the wider zone of a small percentage of clefts in addition to at least one zonula occludens. Goniometric tilting of the specimen in steps of 5 degrees perpendicular to the plane of the wide zone showed that such “linkers” can be revealed in at least 40% of all clefts. They become visible at some tilt angles, although the same area of the cleft is featureless at other angles. Single linker spacing measurements were obtained using a computerized image analysis system, and compiled in a frequency distribution chart. On the basis of these data, two models of a regular linker distribution within the cleft are illustrated. Our results provide evidence for the presence of regularly spaced, cleft-spanning structures within the interendothelial cleft which may have implications for endothelial cell-cell adhesion and permeability.

Using immunohistochemical techniques both at light and electron microscopic levels, the arrangement and distribution of intermediate filaments in Sertoli cells of normal testis (in rat and human), during pre- and postnatal development (in rabbit, rat, and mouse) and under experimental and pathological conditions (human, rat), have been studied and related to the pertinent literature. Intermediate filaments are centered around the nucleus, where they apparently terminate in the nuclear envelope providing a perinuclear stable core area. From this area they radiate to the plasma membranes; apically often a close association with microtubules is seen. Basally, direct contacts of the filaments with focal adhesions occur, while the relationship to the different junctions of Sertoli cells is only incompletely elucidated. In the rat (not in human) a group of filaments is closely associated with the ectoplasmic specializations surrounding the head of elongating spermatids. Both in rat and human, changes in cell shape during the spermatogenic cycle are associated with a redistribution of intermediate filaments. As inferred from in vitro studies reported in the literature, these changes are at least partly hormone-dependent (vimentin phosphorylation subsequent to FSH stimulation) and influenced by local factors (basal lamina, germ cells). Intermediate filaments, therefore, are suggested to be involved in the hormone-dependent mechanical integration of exogenous and endogenous cell shaping forces. They permit a cycle-dependent compartmentation of the Sertoli cell into a perinuclear stable zone and a peripheral trafficking zone with fluctuating shape. The latter is important with respect to the germ cell-supporting surface of the cell which seems to limit the spermatogenetic potential of the male gonad.

The functional loss of mitochondria represents an inherent part in modern theories trying to explain the cutaneous aging process. The present study shows significant age-dependent differences in mitochondrial function of keratinocytes isolated from skin biopsies of young and old donors. Our data let us postulate that energy metabolism shifts to a predominantly non-mitochondrial pathway and is therefore functionally anaerobic with advancing age. CoQ10 positively influences the age-affected cellular metabolism and enables to combat signs of aging starting at the cellular level. As a consequence topical application of CoQ10 is beneficial for human skin as it rapidly improves mitochondrial function in skin in vivo.

Previous studies have shown that many if not most copepods are omnivores and that all zooplankton experience substantial fluctuation in the abundance of food re- sources. We attempt to integrate these two observations by determining the implications of fluctuations in both the abundance and type of available food resources for the survival and reproduction of the omnivorous calanoid copepod Epischura lacustris. Egg production and survivorship were measured in laboratory experiments as food type (zooplankton and phytoplankton) and concentration were experimentally manipulated. In addition, the re- productive activity of a field population was monitored as the abundance and composition of the food resources varied through the year. There are three main results. (1) The alga Cryptomonas erosa and the rotifer Euchlanis dilatata both, when offered alone, serve as adequate foods to support survival and repro- duction of Epischura. (2) However, survival and reproduction of individuals feeding on Cryptomonas vary temporally, even when resource availability is held constant. Extended survival and high egg production rates in the spring are followed by poor survival and minimal egg production rates later in the year. In contrast, survival and reproduction of individuals given the rotifer do not vary significantly across the seasons. (3) Field mea- surements suggest that Epischura lacustris reproduction in situ may depend upon relatively large or evasive animal prey, such as the cladoceran Holopedium gibberum and the rotifer Polyarthra remata.

Summary A 4-month-old female domestic shorthair cat was infected by a virus of the Poxvirus family. The animal developed a severe pneumonia and generalized ulcerating lesions of the skin. Histologically, typical eosinophilic intracytoplasmic inclusion bodies indicative of an Orthopoxvirus (OPV) infection were present. The lung showed grey-white to haemorrhagic nodular lesions with a central zone of complete necrosis of alveolar and bronchial tissue. Electron microscopy from skin and lung nodules revealed typical square-shaped OPV particles. Cultivation of the virus on chorio-allantoic membranes of embryonated chicken eggs resulted in haemorrhagic plaques. Restriction enzyme analysis, PCR and sequencing of the D8L gene identified the OPV isolate as a typical Cowpox virus. It was transmitted by the cat to a human contact person who developed a local nodular dermatitis at the inoculation site in association with signs of general infection and had an increase of OPV-specific neutralizing antibodies in paired serum samples.

Candida endophthalmitis accounts for the majority of fungal endophthalmitis. Despite its clinical relevance there are no controlled trials on different treatment regimens. We report a case of endogenous endophthalmitis caused by azole-resistant Candida albicans following abdominal surgery in an otherwise healthy woman, and review the literature concerning treatment recommendations. In consideration of the serious outcome with loss of sight in insufficiently treated endophthalmitis we like to increase awareness to this disease entity and the possibility of azole-resistance, even in treatment-naive patients.

Objective: To study the pattern and cell type-specificity of collagenase 3, membrane-type 1 matrix metalloproteinase (MT1-MMP), and gelatinase A mRNA expression in the synovial membrane in rheumatoid arthritis (RA). Methods: The mRNA expression of collagenase 3, MT1-MMP, and gelatinase A was characterised by northern blot analysis, reverse transcriptase-polymerase chain reaction, and in situ hybridisation. In situ hybridisation was performed in combination with the immunohistochemical detection of cell type-specific antigens. Results: Synovial membrane specimens from 19 of 21 patients with RA expressing collagenase 3 mRNA were positive for MT1-MMP and gelatinase A mRNA. In control samples from patients without destructive inflammatory joint diseases collagenase 3 mRNA was not expressed and only in two of seven cases was a coexpression of MT1-MMP and gelatinase A mRNA detected. Fibroblast-like cells of the synovial membrane were found to be the predominant source of collagenase 3, MT1-MMP, and gelatinase A mRNA expression in lining and sublining layers as well as at the synovial membrane-cartilage interface. Additionally, the expression of MT1-MMP mRNA was detected in endothelial cells. Collagenase 3 mRNA expression was found in about 5% of CD68 positive macrophages. Conclusions: Collagenase 3 mRNA is expressed simultaneously with MT1-MMP and gelatinase A mRNA in fibroblast-like cells of the synovial membrane in RA. These results suggest (a) a broad extracellular proteolytic potential of fibroblast-like cells and (b) an important role of cell surface associated procollagenase 3 activation by MT1-MMP and gelatinase A for cartilage degradation by invading fibroblast-like cells.

Acquisition of sparingly soluble P sources was investigated in Solarium tuberosum cv. Desiree and in transgenic potato plants with sense (Sp-myc) or antisense expression (αSP) of a sucrose transporter from Spinacia oleracea L., which exhibit increased (Sp-myc) or reduced (αSP) shoot to root translocation of sucrose. Hydroponic culture experiments revealed no genotypic differences in biomass, root-secretory acid phosphatase (APase) activity, utilization of Ca/Mg-phytate, and only marginal exudation of citrate in Sp-myc and cv. D6sir6e plants. However, in soil culture using a Luvisol with organic P as a dominant P fraction, the Sp-myc genotype exhibited increased root dry matter production, which was reverted by antisense inhibition of the sucrose transporter (αSP). In contrast, biomass production in Sp-myc was not enhanced when mineral soils with Ca-P or Fe/Al-P were used for plant cultivation. In all treatments, only marginal mycorrhizal (AM) colonization was observed. The different genotypes exhibited similar activity levels of acid phosphatase in the rhizosphere. Accordingly, mainly bicabonate-extractable organic P was depleted in the rhizosphere soil of all investigated genotypes to a similar extent. The results suggest that genotypic differences in growth response cannot be attributed to variations in release of P-mobilizing root exudates. Enhanced carbohydrate supply to the roots of the Sp-myc genotype may promote the exploitation of a larger soil volume by improved root growth, and thereby phosphatase-mediated P acquisition from organic P sources.

Animal experiments were carried out to investigate whether a protective effect can be achieved in endotoxemia by intravenous (i.v.) application of a polyclonal immunoglobulin preparation (IVIG-IgG/A/M) enriched with 12% IgM and 12% IgA. Following administration of IVIG-IgG/A/M (500 mg/kg), endotoxemia was induced by intraperitoneal inoculation of a sub-lethal dose (5×108 CFU/kg) of Escherichia coli (E. coli) and subsequent i.v. administration of an antimicrobial agent (Imipenem). Plasma endotoxin activity, IL-6 activity, mean arterial pressure, and skeletal muscle oxygen pressure (tpO2) were measured at regular intervals over a total observation period of 7 h. Prophylactic administration of IVIG-IgG/A/M was found to significantly attenuate (P

The usefulness of two nucleic acid detection systems in suspected cases of spontaneous canine herpesvirus (CHV) infection in puppies was evaluated. Formalin-fixed, paraffin-embedded tissues from seven 1–3-week-old naturally infected puppies with lesions characteristic of CHV infection were investigated in a retrospective study. Nested polymerase chain reaction (PCR) and nonradioactive in situ hybridization (ISH) were used to detect nucleotide sequences of the CHV thymidine kinase (TK) gene. According to the original necropsy reports, CHV was isolated in four of the seven puppies using primary canine lung and/or kidney cells. In all seven puppies, gross and histologic lesions consisted of disseminated focal necroses and hemorrhages predominantly in kidneys, lung, liver, and spleen. In addition, few small amphophilic intranuclear inclusion bodies were detected by light microscopy mainly in epithelial cells of kidney, lung, and liver. ISH was performed with a 111-base-pair (bp) digoxigenin-labeled double-stranded DNA probe. Viral DNA was detected in the nuclei of cells near and within lesions. Various cell types, including bronchiolar and alveolar epithelial cells, hepatocytes, renal tubular epithelial cells, neurons, fibrocytes, cardiac myocytes, and endothelial cells, were positive for viral DNA. PCR amplification products of the expected length of 168 bp containing the expected cleavage site for the restriction enzyme EcoRI, derived from paraffin blocks containing lung, kidney, and liver tissues, were detected in all seven puppies. The specificity of the obtained amplicon was further confirmed by Southern blot analysis. ISH and PCR are both useful methods for diagnosing CHV infection in formalin-fixed, paraffin-embedded tissues and are highly specific and sensitive methods for further investigations of the pathogenesis of CHV-induced lesions.

Renal coccidiosis was diagnosed in four bats of different species ( Pipistrellus pipistrellus, Myotis mystacinus, M. nattereri. and Nyctalus noctula). Multiple white and partly indented foci up to 2 mm in diameter were visible on the renal surface. Histologically, the foci appeared as cystic dilated tubules with proliferated epithelium. Asexual and sexual coccidian stages were seen in the epithelial cells, and the extremely distended tubular lumina were filled with schizonts, free zoites, microgamonts, macrogamonts, and unsporulated oocysts. Because the majority of the renal tissue appeared uninvolved in the disease process at the gross and histologic levels and there was no evidence for uremia in other organs, renal function was probably not impaired. Precise classification of the coccidia was impossible because no sporulated oocysts were available. The parasite morphology and the hitherto unreported cystic dilatation of infected tubules containing all developmental stages differ from renal coccidioses reported previously and therefore suggest an undescribed coccidian species.