Herpesviral Hematopoietic Necrosis in Goldfish in Switzerland: Early Lesions in Clinically Normal Goldfish (Carassius auratus).

Cyprinid herpesvirus 2 is a pathogen of goldfish, inducing a disease referred to as herpesviral hematopoietic necrosis. The disease is described so far in Japan, North America, Taiwan, Australia, the United Kingdom, and recently also Italy. Here the authors describe histologic lesions in clinically affected fish in comparison with clinically normal but virus DNA-positive goldfish in Switzerland. While necrosis or enhanced single-cell necrosis in the hematopoietic tissue in the pronephros or mesonephros was evident in dead and sick animals, in clinically normal goldfish, only single-cell necrosis was observed. Virus DNA was demonstrated in dead as well as clinically affected and subclinically infected goldfish by polymerase chain reaction and in situ hybridization. This study identifies the presence of goldfish herpesvirus in Switzerland and highlights the fact that the virus might be more widespread than assumed, as clinically normal goldfish can also carry cyprinid herpesvirus 2, showing histologically similar lesions but of lesser extent and severity.

Epistatic interaction of genetic depression risk variants in the human subgenual cingulate cortex during memory encoding.

Recent genome-wide association studies have pointed to single-nucleotide polymorphisms (SNPs) in genes encoding the neuronal calcium channel CaV1.2 (CACNA1C; rs1006737) and the presynaptic active zone protein Piccolo (PCLO; rs2522833) as risk factors for affective disorders, particularly major depression. Previous neuroimaging studies of depression-related endophenotypes have highlighted the role of the subgenual cingulate cortex (CG25) in negative mood and depressive psychopathology. Here, we aimed to assess how recently associated PCLO and CACNA1C depression risk alleles jointly affect memory-related CG25 activity as an intermediate phenotype in clinically healthy humans. To investigate the combined effects of rs1006737 and rs2522833 on the CG25 response, we conducted three functional magnetic resonance imaging studies of episodic memory formation in three independent cohorts (N=79, 300, 113). An epistatic interaction of PCLO and CACNA1C risk alleles in CG25 during memory encoding was observed in all groups, with carriers of no risk allele and of both risk alleles showing higher CG25 activation during encoding when compared with carriers of only one risk allele. Moreover, PCLO risk allele carriers showed lower memory performance and reduced encoding-related hippocampal activation. In summary, our results point to region-specific epistatic effects of PCLO and CACNA1C risk variants in CG25, potentially related to episodic memory. Our data further suggest that genetic risk factors on the SNP level do not necessarily have additive effects but may show complex interactions. Such epistatic interactions might contribute to the 'missing heritability' of complex phenotypes.

Isolation and partial characterization of a novel virus from different carp species suffering gill necrosis - ultrastructure and morphogenesis.

Two isolates of a novel enveloped RNA virus were obtained from carp and koi carp with gill necrosis. Both isolates behaved identically and could be propagated in different cyprinid cell lines forming large syncytia. The virus was sensitive to lipid solvents and neither exhibited haemadsorption/haemagglutination nor reverse transcriptase activity. Mature virus particles displayed a spherical shape with diameter of 100-350 nm after negative staining and 100-300 nm in ultrathin sections, covered by short projections of 8-10 nm in length. Maturation of virus progeny was shown to occur by budding and envelopment of the filamentous helical nucleocapsids at the cell surface. A detailed comparison of ultrastructure and morphogenesis of the novel virus isolates with selected arena-, ortho- and paramyxoviruses as possible candidates for evaluation of taxonomic classification yielded no consistency in all phenotypic features. Thus, on the basis of ultrastructure the novel virus isolates could not be assigned unequivocally to any established virus family.

Major capsid protein gene sequence analysis of the Santee-Cooper ranaviruses DFV, GV6, and LMBV.

The Santee-Cooper ranaviruses doctor fish virus (DFV), guppy virus 6 (GV6), and largemouth bass virus (LMBV) are members of the genus Ranavirus within the family Iridoviridae. The major capsid protein (MCP) is a main structural protein of iridoviruses and supports the differentiation and classification of ranaviruses. Presently the complete sequence of the MCP gene is known for most ranaviruses with the exception of the Santee-Cooper ranaviruses. In the present study, the complete nucleotide sequence of the MCP gene of DFV, GV6, and LMBV was determined. DFV and GV6 are identical within the MCP gene sequence. The identity compared to the corresponding sequence in LMBV amounts to 99.21%. The MCP gene of DFV, GV6, and LMBV exhibits only approximately 78% identity compared to the respective gene of other ranaviruses. Based on the sequence data obtained in the present study, a Rana MCP polymerase chain reaction (PCR) and subsequent restriction fragment length polymorphism (RFLP) analysis were developed to identify and differentiate ranaviruses, including DFV, GV6, and LMBV.

Ranavirus phylogeny and differentiation based on major capsid protein, DNA polymerase and neurofilament triplet H1-like protein genes.

In this study, we developed new methods for differentiation of ranaviruses based on polymerase chain reaction and restriction enzyme analysis of DNA polymerase and neurofilament triplet H1-like (NF-H1) protein gene. Using these methods, we were able to differentiate the 6 known ranaviruses--Bohle iridovirus (BIV), European catfish virus (ECV), epizootic haematopoietic necrosis virus (EHNV), European sheatfish virus (ESV), frog virus 3 (FV3) and Singapore grouper iridovirus (SGIV)--with 3 less characterised virus isolates: short-finned eel ranavirus (SERV), Rana esculenta virus Italy 282/I02 (REV 282/I02) and pike-perch iridovirus (PPIV). Doctor fish virus (DFV) and guppy virus 6 (GV6) were distinguished as a group from the other viruses. In addition, all 11 isolates were analysed and compared based on nucleotide sequences from 3 different genomic regions: major capsid protein (MCP), DNA polymerase and NF-H1. The partial DNA polymerase gene was sequenced from all analysed viruses. The complete sequence of the MCP and a fragment of the NF-H1 gene were obtained from BIV, ECV, EHNV, ESV, FV3, PPIV, REV 282/I02 and SERV. With the exception of GV6, DFV and SGIV, the sequence analyses showed only a few variations within the analysed viruses. The sequence data suggest that PPIV, REV 282/I02 and SERV are new members of the genus Ranavirus. The methods developed in this study provide tools to differentiate between closely related ranaviruses of different host and geographical origin.

Lung computed tomography density distribution in a porcine model of one-lung ventilation.

BACKGROUND: One-lung ventilation (OLV) exposes the dependent lung to increased mechanical stress which may affect the postoperative course. This study evaluates regional pulmonary gas/tissue distribution in a porcine model of OLV. METHODS: Nine anaesthetized and mechanically ventilated (V(T)=10 ml kg(-1), FI(O(2))=0.40, PEEP=5 cm H(2)O) pigs were studied. After lung separation by an endobronchial blocker, lateral thoracotomy and OLV were performed in six pigs. Three animals served as controls. Static end-expiratory and end-inspiratory spiral computed tomography (CT) scans were done before, during, and after OLV and at corresponding times in controls. CT images were analysed by defined regions of interest and summarized voxels were classified by defined lung X-ray density intervals (atelectasis, poorly aerated, normally aerated, and overaerated). RESULTS: Dependent lungs contained poorly aerated regions and atelectasis with a significant tidal recruitment during conventional two-lung ventilation (TLV) before OLV (expiration vs inspiration: atelectasis 29% vs 14%; poorly aerated 66% vs 44%; normally aerated 4% vs 41% of the dependent lung volume, P<0.05). During OLV (V(T)=10 ml kg(-1)), cyclic recruitment was increased. The density spectrum of the ventilated lung changed from consolidation to aeration (expiration vs inspiration: atelectasis 10% vs 2%; poorly aerated 71% vs 18%; normally aerated 19% vs 79%, P<0.05). After OLV, increased aeration remained with less atelectasis and poorly aerated regions. Lung density distribution in the non-dependent lung of OLV pigs was unaltered after the period of complete lung collapse. CONCLUSIONS: Cyclic tidal recruitment during OLV in pigs was associated with a persistent increase of aeration in the dependent lung.

Fish rhabdoviruses: molecular epidemiology and evolution.

Rhabdoviruses may cause serious diseases in wild and farmed fish. Within the Rhabdoviridae six genera have been established: Ephemerovirus, Cytorhabdovirus, Nucleorhabdovirus, Lyssavirus, Vesiculovirus, and Novirhabdovirus. Viruses that infect fish are official or tentative members of the genera Vesiculovirus and Novirhabdovirus, or are listed as unassigned rhabdoviruses. In this report, we summarize and discuss published and our own unpublished data on the molecular epidemiology and phylogeography of fish rhabdoviruses including intrapopulational differences and subgrouping of fish rhabdoviruses, in particular the species spring viremia of carp virus (SVCV), infectious hematopoietic necrosis virus (IHNV) and viral hemorrhagic septicemia virus (VHSV).

Stridulatory pattern generation in acridid grasshoppers: metathoracic interneurons in Stenobothrus rubicundus (Germar 1817).

Stridulation was elicited in tethered gomphocerine grasshoppers of the species Stenobothrus rubicundus in order to identify interneurons of the stridulation pattern generator, and describe their morphological and physiological properties. Nine types of such neurons could be found and characterized; eight of those could additionally be compared to corresponding neuron types previously known from other species. As shown in detail for one selected type, the neurons of the stridulation pattern generator are very similar in their anatomical appearance, and possess similar physiological qualities at least in two species with similar stridulation patterns. Stridulation interneurons of species with largely different stridulatory motor patterns have a similar morphology, but show a different activation timing throughout the stridulation. Nevertheless, special properties such as resetting or initiation capability of certain stridulation interneurons seem to be conserved throughout the species. The results suggest that the stridulation pattern generator of different species consists of a uniform set of interneurons that change their activity pattern to produce species-specific song movements.

Walk with a pen, or fly with a computer. Can electronic dentistry work for you?

In the last several years, dentistry has crossed over into the new frontier of electronic dentistry. It has embraced such developments as computer programs for producing digital radiographs and photographs, as well as digital programs that enhance these images, store and organize them into a retrievable "chart-like" fashion, and transmit them via the Internet. In Europe, I saw patients with an electronic "health card." This credit card-sized CD can carry all the information on a patient's written charts, results of laboratory tests, radiographic/imaging information and more. It is expected that the mobile phone will be an alternate vehicle for patient records, and that these records will be accessed with a password security system. This will allow patients to carry their records from location to location. Certainly, the dental implications of such seemingly advanced processes are evident. The expression, "The future is now," was never truer.

Complete genomic sequence of viral hemorrhagic septicemia virus, a fish rhabdovirus.

The complete nucleotide sequence of the fish rhabdovirus viral hemorrhagic septicemia virus (VHSV) has been determined. The genome comprises 11158 bases and contains six long open reading frames encoding the nucleoprotein N, phosphoprotein P, matrix protein M, glycoprotein G, nonstructural viral protein NV, and polymerase L. Genes are arranged in the order 3'-N-P-M-G-NV-L-5'. The exact 3' and 5' ends were determined after RNA-oligonucleotide ligation or RACE. They show inverse complementarity as in other rhabdovirus genomes. Nucleotide and deduced amino acid sequences exhibit significant homology to corresponding sequences in the related fish rhabdovirus infectious hematopoietic necrosis virus.

Identification of group VS116 strains among Borrelia burgdorferi sensu lato grown from the hard tick, Ixodes ricinus (Linnaeus, 1758) by off-coupled restriction fragment length polymorphism analysis.

A total of 67 spirochetal isolates grown from the hard tick, Ixodes ricinus were analysed by PCR amplification of the spacer region between two conserved structures, the 3' end of the 5S rRNA and the 5' end of the 23S rRNA genes of Borrelia burgdorferi sensu lato. A 246-255 bp amplicon was generated from 13 reference strains of B. burgdorferi sensu lato representing the three major genospecies, B. burgdorferi sensu stricto, B. garinii, and B. afzelii, and from all 67 spirochetal isolates from ticks but not from B. hermsii. As could be confirmed by DNA sequence analysis, restriction fragment length polymorphism (RFLP) analysis of the PCR product after cleavage with DraI and MseI distinguished between the three major genospecies: out of the 67 B. burgdorferi sensu lato isolates from ticks, 27 (40.3%) were typed as B. burgdorferi sensu stricto including five isolates with a unique DraI or MseI pattern. 26 isolates (38.8%) were typed as B. garinii and 6 (9.0%) as B. afzelii, respectively. A group of eight isolates (11.9%) displayed a unique MseI pattern identical to that described for a putative new European genospecies of Borrelia burgdorferi sensu lato designated VS116. DNA sequences of the PCR product of seven of these isolates tested were by less than 88.5% identical with the established European major genospecies but shared 98% to 100% homology with that of database-derived sequences of strain VS116 from Switzerland and strain UK from England which are both representatives of the European genomic group VS116.

RAP1-like binding activity in islet cells corresponds to members of the Sp1 family of transcription factors.

Deletion and mutational analyses of the gastrin promoter have identified a binding site for the yeast transcription factor RAP1 relevant for transcriptional activation in islet cells. We here report that the mammalian transcription factors binding to this site in islet cells are the Sp transcription factor members Sp1 and Sp3. Furthermore, functional analyses revealed Sp1- and Sp3-mediated transcriptional activation of gastrin. These data reveal that the zinc finger proteins Sp1 and Sp3 do have similar binding specificities as the multifunctional yeast RAP1 protein.

Identification of the non-virion (NV) protein of fish rhabdoviruses viral haemorrhagic septicaemia virus and infectious haematopoietic necrosis virus.

Sequence analysis of a 795 nucleotide region of the fish rhabdovirus viral haemorrhagic septicaemia virus (VHSV) genome revealed one complete and one partial ORF of 369 and 153 nucleotides, respectively. The latter ORF probably encodes the amino-terminal part of the L (polymerase) protein. The former ORF potentially encodes a 122 amino acid protein. The location of this ORF as well as the size and deduced structure of the translation product indicate that it represents a homologue of the non-virion (NV) protein of the related infectious haematopoietic necrosis virus (IHNV). Antisera raised against prokaryotically expressed NV protein of VHSV and IHNV were used to detect NV expression in VHSV- and IHNV-infected cells by Western Blot and immunofluorescence analyses. We present here the sequence of the VHSV NV gene and demonstrate the presence of IHNV and VHSV NV proteins in virus-infected cells.

Complete genomic sequence of the fish rhabdovirus infectious haematopoietic necrosis virus.

The complete nucleotide sequence of the genome of the fish rhabdovirus infectious haematopoietic necrosis virus (IHNV) has been determined after cDNA cloning of the viral genomic RNA. Sequence analysis showed the presence of six open reading frames encoding the nucleoprotein N, the matrix proteins M1 and M2, the glycoprotein G, a so-called non-structural protein NV, and the RNA polymerase L. The genome organization is 3'N-M1-M2-G-NV-L 5'. The extreme 5' and 3' ends of the genome were sequenced after RNA ligation or RACE. Prokaryotic expression products of the open reading frames predicted to encode the matrix proteins M1 and M2, the glycoprotein G and the NV protein reacted with rabbit anti-IHNV serum thereby confirming their identity. This is the first complete nucleotide sequence of a fish rhabdovirus. Knowledge of the complete sequence is an essential prerequisite for future manipulation of the genome and also serves to provide gene- and protein specific reagents for use in further examination of the replication of the fish rhabdoviruses.

The iron-binding protein ferritin is expressed in cells of the osteoblastic lineage in vitro and in vivo.

Ferritin, a metal-binding protein responsible for maintaining the bioavailability of iron, has been demonstrated in cells of the osteoblastic lineage. Messenger RNAs encoding the light and heavy chain subunits of ferritin were detected in ROS 17/2.8, ROS 25/1, and UMR106 rat osteosarcoma cell lines, in fetal rat calvaria, and in primary cultures of rat calvarial osteoblast-like cells. In vivo, the expression of ferritin light-chain mRNA was observed in both active osteoblasts and in osteocytes. A 450-kD iron-binding protein was immunoprecipitated from ROS 17/2.8 cells by an antiferritin antiserum. This protein comigrated with native ferritin, and could be dissociated into subunits comigrating with ferritin light and heavy chains. Addition of extracellular Fe59-transferrin to cultures of ROS 17/2.8 cells resulted in the sequestration of the iron in intracellular ferritin. These observations demonstrate that cells of the osteoblastic lineage possess a functional ferritin-based iron uptake and storage system capable of regulating metal homeostasis in bone.

Mesothelioma-like tumors of the pleura: a review of 72 autopsy cases.

The 72 mesothelioma-like tumors of the pleura (MLTP) found among 33 500 autopsy cases collected over more than 30 years are reviewed. MLTP have a worse prognosis than the 106 cases of pleural mesothelioma autopsied in our institutes with regard to survival time and metastatic spread. In MLTP, adenocarcinomas predominate with a wide range of histological and cytological variation and prominent development of connective tissue having its origin in the periphery of the lung. These intrapulmonary primary tumors often fulfill the criteria of pulmonary scar cancer. Etiologically, there is no correlation between the origin of this tumor and smoking or exposure to asbestos. The absence of mucus formation and glandular differentiation, together with the presence of spindle-shaped carcinoma components and strong mesothelial or stroma proliferation, can make the differential diagnosis between this tumor type and mesothelioma difficult. Immunohistological investigations were performed on 11 cases with antibodies against intermediate filament proteins, vascular endothelium, collagen IV, macrophage antigens, carcinoembryonic antigen (CEA), LeuM1, and the antibody BerEP4. Our investigation shows that a battery of several tumor markers, such as antibodies against LeuM1, CEA, and the antibody BerEP4, as well as staining with periodic acid/Schiff/diastase discriminate primary from secondary pleural neoplasms, whilst intermediate filament proteins alone are of little diagnostic value.

Higher capillary filtration rate in the calves of endurance-trained subjects during orthostatic stress.

The effect of endurance training on the rate of transcapillary filtration during orthostasis was studied in the human calf. Two groups of sports students with markedly different aerobic capacities performed an orthostatic tilt table test (25 min supine, 10 min upright, 10 min supine). The following parameters were measured: heart rate, brachial and peripheral blood pressure (Finapres), calf volume changes (impedance), and calf blood flow (venous-occlusion-technique). The two groups did not differ in maximal calf circumference, body height, or weight. No syncope occurred, and heart rate and blood pressure responses to upright tilt were similar in both groups. However, the capillary filtration rate revealed much higher values in the trained group: 0.086 vs. 0.036 ml.min-1.100 ml-1. The estimated additional fluid accumulation in the interstitial space in trained subjects may be as high as 260 ml within the first 20 min of orthostasis and may play a role in often reported late syncopes, depending on the preexisting fluid state.