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Genome dynamics and diversity of Shigella species, the etiologic agents of bacillary dysentery.

The Shigella bacteria cause bacillary dysentery, which remains a significant threat to public health. The genus status and species classification appear no longer valid, as compelling evidence indicates that Shigella, as well as enteroinvasive Escherichia coli, are derived from multiple origins of E.coli and form a single pathovar. Nevertheless, Shigella dysenteriae serotype 1 causes deadly epidemics but Shigella boydii is restricted to the Indian subcontinent, while Shigella flexneri and Shigella sonnei are prevalent in developing and developed countries respectively. To begin to explain these distinctive epidemiological and pathological features at the genome level, we have carried out comparative genomics on four representative strains. Each of the Shigella genomes includes a virulence plasmid that encodes conserved primary virulence determinants. The Shigella chromosomes share most of their genes with that of E.coli K12 strain MG1655, but each has over 200 pseudogenes, 300 approximately 700 copies of insertion sequence (IS) elements, and numerous deletions, insertions, translocations and inversions. There is extensive diversity of putative virulence genes, mostly acquired via bacteriophage-mediated lateral gene transfer. Hence, via convergent evolution involving gain and loss of functions, through bacteriophage-mediated gene acquisition, IS-mediated DNA rearrangements and formation of pseudogenes, the Shigella spp. became highly specific human pathogens with variable epidemiological and pathological features.

DNA Transposable Elements↗

Characterization of isolates and clones of Leishmania by analysis of kinetoplast DNA.

The genetic characterization of pathogenic isolates of Leishmania was attempted by analysis of the molecular properties of kinetoplast DNA (kDNA) minicircles. Unit minicircle size is not conserved during speciation of Leishmania since the minicircles of strains and clones of L t major are smaller (700 bp) than those found in certain strains of L mexicana ssp (820 bp), L donovani (850 bp) or L t tropica (900 bp). Schizodeme analysis of minicircles reveals a high degree of sequence divergence in kDNA of Leishmania with the degree of microheterogeneity varying between species. This sequence divergence allows the discrimination of species, strains, and clones of Leishmania into schizodemes . Southern blot hybridization experiments reveal that at high stringency overall minicircle sequence homology is conserved among clones and strains of one species (L t major) but not between different species. This property of minicircle DNA permits the use of kDNA probes as a species-specific diagnostic test for the identification of unknown Leishmania isolates. The properties of kDNA from an L t tropica strain LRC- L32 (a " recidiva " organism) are so diverged from those of L t major strains as to support the classification [22, 23] of L t tropica and L t major as separate species of Leishmania rather than subspecies of L tropica.

Animals↗

Biomarker identification through spatial proteomics for the characterization of indeterminate thyroid nodules.

PURPOSE: The identification of novel molecular biomarkers may assist in the characterization of indeterminate thyroid nodules, which pose significant diagnostic challenges. Here, we aimed to explore the potential of proteomic analyses to support biomarker discovery in challenging thyroid lesions. METHODS: Linear Discriminant Analysis (LDA) was applied to Matrix-Assisted Laser Desorption Ionization Mass Spectrometry Imaging (MALDI-MSI) data from 44 thyroid neoplasms to select the most impactful molecular features for the classification of different tumor histologies, as well as for the distinction between NRAS-mutant (mNRAS) and NRAS-wild-type (wtNRAS) tumors. Relevant peaks were subsequently identified through nanoscale liquid chromatography electrospray ionization tandem mass spectrometry (nLC-ESI-MS/MS). RESULTS: The LDA selected nine relevant molecular markers distinguishing noninvasive follicular thyroid neoplasms with papillary-like nuclear features (NIFTPs) from other tumor histologies (balanced accuracy = 73%), as well as 19 relevant markers able to identify mNRAS cases (balanced accuracy = 84%). Nine differentially expressed proteins were putatively identified: among them, ATP-dependent RNA helicase DDX42 showed a similar distribution between NIFTPs and papillary thyroid carcinomas (PTCs) / follicular variant PTCs (FVPTCs), while the distribution of the Histone H4 signal was similar between NIFTPs and follicular adenomas (FAs). In addition, Protein disulfide-isomerase A1 and Complement C4-B were overexpressed in wtNRAS compared to mNRAS cases, regardless of histology. CONCLUSION: The LDA-selected features enable to distinguish NIFTPs from morphologically similar lesions and to discriminate between mNRAS and wtNRAS cases. The identified markers might complement genetic analyses and provide insights into the distinct pathogenic drivers behind the development of mNRAS compared to wtNRAS lesions.

Humans↗

Comparative typing of Pseudomonas species isolated from the aquatic environment in Greece by SDS-PAGE and RAPD analysis.

AIMS: Three broadly used typing methods were employed in order to assess and compare the identification and classification of environmental Pseudomonas strains. The reproducibility, typeability and discriminatory power of the methods were also compared to evaluate their application. Finally, the potential impact on public health of the isolates is to be discussed. METHODS AND RESULTS: Pseudomonas strains (160) isolated from the aquatic environment in Greece and identified by a rapid identification commercially available system (API20NE), were subjected to whole-cell protein electrophoresis (Sodium dodecyl sulfate-polyacrylamide gel electrophoresis) and Randomly Amplified Polymorphic DNAs (RAPD) using two 10-mer primers. In general, the obtained results were in agreement. Twenty isolates that could not be identified by the API20NE system were classified by the other methods. CONCLUSIONS: Rapid identification systems may serve only for a first rough identification of environmental Pseudomonads. In order to acquire further information, so that conclusions about their role in the ecosystem and human health could be drawn, other phenotypic or genotypic methods have to be applied. SIGNIFICANCE AND IMPACT OF STUDY: It is important, from a public health point of view, to monitor the identities of environmental Pseudomonas isolates using specific methods due to their ubiquity, heterogeneity and their pathogenicity, either established or potential.

Bacterial Typing Techniques↗

High frequency of microsatellite instability in intestinal-type gastric cancer in Korean patients.

BACKGROUND: Although there have been some reports on microsatellite alterations in gastric cancer, findings are inconsistent regarding the associations between histological classification and microsatellite instability (MSI). In the present study, we attempted to determine whether Lauren's histological subtypes are related with MSI status. METHODS: Paraffin-embedded tissue samples from 14 diffuse-type and 14 intestinal-type gastric adenocarcinomas were matched up according to patient gender and age. Mononucleotide markers (BAT25 and BAT26) and dinucleotide markers (D2S123, D5S346, and D175S250) were used for MSI analyses. Microsatellite genotypes were categorized in terms of high MSI incidence (MSI-H, >30% positive marker) or low MSI incidence (MSI-L, <30% positive marker). Losses of hMLH1 and hMSH2 protein expression were immunohistochemically studied. RESULTS: MSI-H was observed in 11 cases (78%) of the 14 intestinal-type cases as compared to 3 (21%) of the 14 diffuse-type cases (p=0.007). In MSI-H tumors, 10 cases (71%) showed losses of hMLH1 protein expression, while 2 cases (14%) in MSI-L tumors showed losses of hMLH1 protein expression (p=0.006). CONCLUSION: MSI-H tumors are more frequently found in intestinal-type gastric cancer, which suggests the possibility that there are different pathogenic pathways in gastric carcinogenesis according to histologic type.

Adaptor Proteins, Signal Transducing↗

New approaches to pathogenic gene function discovery with human squamous cell cervical carcinoma by gene ontology.

PURPOSE: This study utilized mRNA differential display and the Gene Ontology (GO) analysis to characterize the multiple interactions of a number of genes with gene expression profile involved in squamous cell cervical carcinoma. METHODS: mRNA differential displays were used to identify potential transcripts that were differentially expressed between cervix cancers of 13 patients (invasive cancer stages Ib-IIb) and universal reference RNAs comprised of 17 different normal cervixes. Aberrant bands were excised and used to make cDNA, which was sequenced. DNA sequences were compared to other nucleic acids in the NCBR database for homology. Transcript expression was verified in select samples using RT-PCR and North blotting. The specific functions were correlated with gene expression patterns via gene ontology. RESULTS: Fifty-eight genes were up- or down-regulated above 2-fold and organized into reciprocally dependent sub-function sets depending on the cervical cancer pathway. The GO analysis showed that squamous cell cervical carcinogenesis underwent complete up-regulation of cell cycle, transport, epidermal differentiation, protein biosynthesis, and RNA metabolism. Also, genes belonging to protein metabolism and catabolism activity were significantly up-regulated. In contrast, significant down-regulation was shown in muscle development, cell adhesion, and damaged DNA binding activity. CONCLUSION: The GO analysis can overcome the complexity of the gene expression profile of the squamous cell cervical carcinoma-associated pathway and identify several cancer-specific cellular processes as well as genes of unknown function. Also, GO analysis can serve as a powerful basis for a molecular classification of carcinogenesis.

Apoptosis↗

[Results from the Central Laboratory for Streptococci Research in Kiel from 1965 to 1977 - a survey (author's transl)].

At the Central Laboratory for Streptococci Research at the Institute for Hygiene of the Federal Dairy Research Centre in Kiel a data bank has been established which comprises as well bibliographical data (until now more than 8000 titles out of the international literature concerning streptococci) as laboratory data of all strains which have been identified since 1965. For this present publication as a survey the total material covering 34,935 strains was analyzed and interpreted. Besides a short introduction describing the general examination procedure for streptococci and the encoding of results for the data bank the material is condensed in different tables considering the incidence of pathogenic species occuring mainly in man and animals. In addition findings of enterococci and lactic streptococci are presented. 41.8% of the strains examined were isolated from human specimens, 46.7% from cattle and the remaining ones from pigs, horses, dogs, laboratory animals and others. Only 2.0% of all cultures could not be identified as one of the known species. Remarkable observations are - the high incidence of group B-Streptococci in man - the unexpected frequency of typical "human" streptococci as those of group F and G and also Sc. milleri and Sc. MG in different animals - the possible interrelation of group L- and P-streptococci in pigs and cattle - the overlap of Sc. faecalis and Sc. faecium in man and animals showing no typical distribution as it is postulated in the literature - the high percentage of unidentifiable streptococci from pigs and laboratory animals as an eventual indication for some new species. In consecuting papers some problems emerging from the presented material and own experiments will be discussed as - interrelation of group B-Streptococci in man and cattle - incidence and importance of enterococci in humans, animals and foodstuffs and - classification of species not to identify by serological grouping.

Animals↗

Candida dubliniensis: an emerging opportunistic pathogen.

The incidence of opportunistic fungal infections continues to increase, partly as a result of the continuing AIDS epidemic. Candida albicans remains the most important fungal pathogen and is frequently associated with oral candidiasis in HIV-infected individuals. Over the past decade, however, there has been an increasing number of reports implicating other Candida species, such as C. tropicalis, C. glabrata and C. krusei, in disease in these patients and in other patient groups. During the same period there have also been frequent reports in the literature describing what have generally been termed "atypical" C. albicans strains. These isolates have usually been recovered from symptomatic HIV-infected individuals and are unidentifiable as any recognized Candida species using conventional criteria. Two such groups of isolates recovered from cases of oral candidiasis in Irish and Australian HIV-infected and AIDS patients have been postulated to constitute a novel species which has been termed C. dubliniensis. These isolates are phenotypically very similar to C. albicans in that they produce germ tubes and chlamydospores. However, they have unusual carbohydrate assimilation patterns and grow poorly or not at all at 42 degrees C. Using a variety of DNA fingerprinting techniques and karyotype analysis, the genomic organization of C. dubliniensis was shown to be distinctly different from that of C. albicans. Classification of C. dubliniensis as a separate species was confirmed by phylogenetic analysis, whereby the comparison of ribosomal RNA sequences demonstrated that C. dubliniensis isolates formed a cluster clearly distinct from other Candida species, including C. albicans, to which it is most closely related. Since its original identification, atypical Candida isolates from around the world have been positively identified as belonging to this species. To date, isolates of C. dubliniensis have been recovered mainly from the oral cavities of HIV-infected individuals and are most frequently implicated in cases of recurrent infection following antifungal drug treatment. The clinical importance of this species and the role of drug resistance in its epidemiology have yet to be determined.

AIDS-Related Opportunistic Infections↗

Sarcocystis in man: a review and report of five cases.

Sarcocystis was identified in biopsy specimens of skeletal muscle from two adults in Singapore and one in Bombay, and in muscle obtained at autopsy from an adult in Uganda, and in the heart of a child in Costa Rica. Review of case reports revealed that in seven reported cases, including three reported by Lindemann, non-parasite objects were misinterpreted as sarcocysts; in four instances organisms of undetermined classification were described, and in 35 cases true sarcocysts were observed. Among the sarcocysts seen in the 40 cases (35 old, 5 new), seven morphological types were recognized, each representing one to several different species, all of which are zoonotic and none of which can be designated Sarcocystis lindemanni. Among the four types of sarcocysts found in skeletal muscle, three closely resembled a corresponding species found commonly in monkeys: one from a man in Uganda corresponding to a species in Ceropithicus talapoin, forms from India resembling one or two species in Macaca mulatta, and forms from Southeast Asia resembling a species in Macaca fascicularis. Among three types of sarcocysts found in the human heart, one resembled a species commonly seen in the heart of cattle. Of the 40 Sarcocystis infections in man, 13 probably were acquired in Southeast Asia, 8 in India, 5 in Central or South America, 4 each in Africa and Europe, 3 in USA, 1 IN China and 2 in unknown localities. Associated conditions include muscle soreness or weakness in 7 cases, subcutaneous swellings in 5, eosinophilia in 2, and periarteritis or polyarteritis nodosa in 2 cases. However, evidence of pathogenicity of the mature sarcocyst is inconclusive.

Adult↗

Bacterial source tracking and shellfish contamination in a coastal catchment.

Introduced pathogens from faecal material can make their way into the aquatic environment from a number of catchment sources. These sources typically include sewage outfalls, seepage from septic tanks, and urban and agricultural runoff. Shellfish as filter feeders are particularly susceptible to contamination in faecally contaminated waters and a range of microbiological indicators have been developed to assess the levels of contamination and likely risks to public health (Hackney and Pierson, 1994). This paper outlines the application of bacterial source tracking (BST) in a shellfish growing area in part of the Port Stephens estuary along the NSW north coast. The approach is based on the premise that bacterial isolates from different faecal sources will have significantly different resistance patterns to the battery of antibiotics and concentrations tested. Faecal streptococci (FS) were isolated from several possible faecal sources: beef and dairy cattle, chickens and humans. The resistance patterns of these isolates to four different concentrations of four antibiotics were compared to those of FS isolates obtained from samples collected upstream and in the vicinity of the oyster leases. Discriminant analysis was performed using the patterns from the known source isolates and the rate of correct classification was determined for each source. The predictive function of discriminant analysis was then used to determine the most probable source of each of the unknown isolates from Tilligerry Creek, the drainage channels to the estuary, and the shellfish leases. Preliminary results are presented here and suggest that there is no single significant source of faecal contamination, rather there are contributions from a range of sources. The findings may have implications for the ways in which land use activities and catchments are managed in similar estuarine locations with a shellfish industry.

Agriculture↗

Nephritogenic lupus antibodies recognize glomerular basement membrane-associated chromatin fragments released from apoptotic intraglomerular cells.

Antibodies to dsDNA represent a classification criterion for systemic lupus erythematosus. Subpopulations of these antibodies are involved in lupus nephritis. No known marker separates nephritogenic from non-nephritogenic anti-dsDNA antibodies. It is not clear whether specificity for glomerular target antigens or intrinsic antibody-affinity for dsDNA or nucleosomes is a critical parameter. Furthermore, it is still controversial whether glomerular target antigen(s) is constituted by nucleosomes or by non-nucleosomal glomerular structures. Previously, we have demonstrated that antibodies eluted from murine nephritic kidneys recognize nucleosomes, but not other glomerular antigens. In this study, we determined the structures that bind nephritogenic autoantibodies in vivo by transmission electron microscopy, immune electron microscopy, and colocalization immune electron microscopy using experimental antibodies to dsDNA, to histones and transcription factors, or to laminin. The data obtained are consistent and point at glomerular basement membrane-associated nucleosomes as target structures for the nephritogenic autoantibodies. Terminal deoxynucleotidyl-transferase-mediated dUTP nick end-labeling or caspase-3 assays demonstrate that lupus nephritis is linked to intraglomerular cell apoptosis. The data suggest that nucleosomes are released by apoptosis and associate with glomerulus basement membranes, which may then be targeted by pathogenic anti-nucleosome antibodies. Thus, apoptotic nucleosomes may represent both inducer and target structures for nephritogenic autoantibodies in systemic lupus erythematosus.

Animals↗

Metabolite profiling of fungi and yeast: from phenotype to metabolome by MS and informatics.

Filamentous fungi and yeast from the genera Saccharomyces, Penicillium, Aspergillus, and Fusarium are well known for their impact on our life as pathogens, involved in food spoilage by degradation or toxin contamination, and also for their wide use in biotechnology for the production of beverages, chemicals, pharmaceuticals, and enzymes. The genomes of these eukaryotic micro-organisms range from about 6000 genes in yeasts (S. cerevisiae) to more than 10,000 genes in filamentous fungi (Aspergillus sp.). Yeast and filamentous fungi are expected to share much of their primary metabolism; therefore much understanding of the central metabolism and regulation in less-studied filamentous fungi can be learned from comparative metabolite profiling and metabolomics of yeast and filamentous fungi. Filamentous fungi also have a very active and diverse secondary metabolism in which many of the additional genes present in fungi, compared with yeast, are likely to be involved. Although the 'blueprint' of a given organism is represented by the genome, its behaviour is expressed as its phenotype, i.e. growth characteristics, cell differentiation, response to the environment, the production of secondary metabolites and enzymes. Therefore the profile of (secondary) metabolites--fungal chemodiversity--is important for functional genomics and in the search for new compounds that may serve as biotechnology products. Fungal chemodiversity is, however, equally efficient for identification and classification of fungi, and hence a powerful tool in fungal taxonomy. In this paper, the use of metabolite profiling is discussed for the identification and classification of yeasts and filamentous fungi, functional analysis or discovery by integration of high performance analytical methodology, efficient data handling techniques and core concepts of species, and intelligent screening. One very efficient approach is direct infusion Mass Spectrometry (diMS) integrated with automated data handling, but a full metabolic picture requires the combination of several different analytical techniques.

Biochemistry↗

Visual Detection and Stratification of Pathogenic mtDNA SNV Heteroplasmy by Balancing FnCas12a Signal Output and Allelic Discrimination.

Assessment of pathogenic mitochondrial DNA (mtDNA) single-nucleotide variant (SNV) heteroplasmy is important for molecular diagnostics, yet rapid visual profiling remains analytically challenging because an assay must combine single-nucleotide allelic discrimination, mutant-fraction-associated readout, and suitable target access. Herein, we report VISTA (visual identification and stratification of targeted mtDNA alleles), a broad-PAM FnCas12a assay that rebalances trans-cleavage signal output and mutant-wild-type discrimination for visual mtDNA SNV heteroplasmy analysis. VISTA uses unmodified FnCas12a with relaxed TTN PAM recognition and integrates crRNA spacer-length engineering with PEG8000/acBSA reaction tuning to improve the practical signal-discrimination balance without nuclease engineering. At the m.3243A>G model locus, spacer truncation enhanced mutant-wild-type discrimination, while molecular-dynamics simulations identified spacer-dependent differences between matched and mismatched complexes at the crRNA-DNA interface. The optimized assay resolved defined synthetic m.3243A>G heteroplasmy gradients by fluorescence imaging and was further adapted to lateral-flow detection. In locus-specific analyses of a deidentified collection of 74 peripheral-blood samples, fluorescence and lateral-flow readouts achieved ROC AUC values above 0.9 for mutant-allele classification after target-region amplification. Fluorescence supported heteroplasmy-associated profiling, whereas lateral flow provided a visual, semiquantitative readout for relative ranking based on the T/C ratio rather than absolute heteroplasmy measurement. VISTA therefore provides an accessible dual-readout analytical strategy for visual detection and heteroplasmy-associated profiling by tuning the FnCas12a signal output and allelic discrimination.

DNA, Mitochondrial↗

Phylogenetic tree and sequence similarity of beta-lactamases.

beta-Lactamases are the main cause of beta-lactam resistance in many pathogenic bacteria. These enzymes can be detected in a variety of pathogenic as well as non-pathogenic bacteria. The cyanobacteria are also known to produce a beta-lactamase. Recently, the amino acid sequences and the three-dimensional structures of some of these beta-lactamases have been clarified. On the basis of the amino acid sequences of 47 beta-lactamases and the computer-aided analysis, a phylogenetic tree is proposed in this paper. According to the tree, beta-lactamases are classified into six groups. Group 1 beta-lactamases are mainly composed of plasmid-mediated enzymes from gram-negative bacteria. However, chromosome-derived beta-lactamases from Klebsiella pneumoniae and Rhodopseudomonas capsulata take part in this group. Group 2 enzymes consist of a part of the chromosome-encoded beta-lactamases from Streptomyces, and chromosome-mediated enzymes from Yersinia enterocolitica, Citrobacter diversus, and Klebsiella oxytoca. Chromosome-encoded beta-lactamases from gram-negative bacteria form group 3. Group 4 is composed of metalloenzymes, whereas group 5 consists of OXA type beta-lactamases. Chromosome-encoded beta-lactamases from gram-positive bacteria form group 6. Comparison of the amino acid sequences among these groups confirmed the phylogenetic tree and the classification: the beta-lactamases in each group have its particular conserved amino acid sequences. In addition, the tree provides more detailed classification and time-scale mutual relationships and predicts new types of beta-lactamases that may be found. Furthermore, the classification deduced from the tree is generally in accord with the one based on the amino acid sequences reported previously. However, the class A beta-lactamases are clearly divided into three groups: groups 1, 2, and 6. RDF2 analysis shows that some combinations between beta-lactamases and beta-lactam-interacting proteins as well as eukaryotic proteins have a low but significant evolutionary relatedness.

Amino Acid Sequence↗

Investigations on the clonality of isolates of Pasteurella gallinarum obtained from turkeys in Germany.

Pasteurella gallinarum has been considered an opportunistic pathogen rather than a primary pathogen for chickens. As P. gallinarum has been found to have a high genotypic diversity, one would expect a polyclonal distribution among isolates from different farms if this organism is a secondary invader. The aims of this investigation were to genetically characterize isolates obtained from outbreaks affecting several turkey farms to confirm the existence of the infection in turkeys and to investigate the genetic relationship between isolates from affected farms. A total of 17 isolates from 14 outbreaks of respiratory disease in Germany were subjected to extended phenotypic and genotypic characterization. All isolates were of the same phenotype, typical of P. gallinarum. Ribotyping of three isolates using either HpaII or HindIII showed that they had identical profiles and indicated that the isolates all originated from the same clone. Comparison with HpaII ribotypes from a previous study showed that the pattern was identical to that obtained with isolates from a Zimbabwean outbreak in chickens during 1999 to 2000. Restriction endonuclease analysis typing of 14 isolates from all 14 farms showed that they had identical profiles but these differed from those obtained with isolates from the Zimbabwean outbreak. Sequencing of the 16S rRNA gene and sequence comparisons with other Pasteurellaceae confirmed their classification as P. gallinarum. Identification of the same clone of P. gallinarum from 14 outbreaks of acute respiratory disease in turkeys within a time period of 2 months suggests a common source of infection, and that P. gallinarum probably played a primary role rather than a secondary role in the outbreaks.

Animals↗

[The state of knowledge of the class of trematodes and the chief problems in studying them].

The paper deals with the current state of knowledge of Trematoda, trends and aims of further investigations. Up to now the classification system of trematodes has been unsatisfactory though for several large groups it is elaborated with due regard to the biology and ecology of these worms. The main deficiency of the present-day taxonomy of trematodes consists in the existence of two separate classifications, for adult worms and for their larvae (cercariae). Among the existing systems the classification suggested by Odening (1960-1963) is of the greatest interest. We do not agree with his division of the class into three subclasses (including Aspidogastrea) and Diginea into two subclasses, "Sporocystoinei" and "Redioinei", but follow the composition of orders established by him. Great difficulties arise in establishing the characters, taxonomic criteria of trematodes. However, a number of characters can be suggested for differentiation of species and genera at various developmental stages. Deficiency in the taxonomy of these worms is due to a wrong evaluation of their characters and to the lack of common collection and treatment techniques. On describing species one should proceed, if possible, from the whole complex of morphological and biological peculiarities of different ontogenetic stages of the forms under study. A study of the ultrafine structure, hysto- and biochemistry of trematodes will enable us to associate better the morphology and the function of organs and to go deeper into the functional morphology and functional biochemistry. The main task of further investigations is closely connected with the elaboration of a natural system of Trematoda, a study of diverse and deep interrelations in the "parasite-host" system and with the development of scientific methods of control of pathogenic species on the biological basis.

Animals↗

Overview on chronic viral cardiomyopathy/chronic myocarditis.

Myocarditis is most often induced by cardiotropic viruses and often resolves with minimal cardiac remodelling and without discernable prognostic impact. Acute myocarditis has a highly diverse clinical presentation (asymptomatic, infarct-like presentation, atrioventricular (AV)-block, atrial fibrillation, sudden death due to ventricular tachycardia, fulminant myocarditis with severely depressed contractility). Progression of myocarditis to its sequela, dilated cardiomyopathy (DCM), has been documented in 20% of cases and is pathogenically linked to chronic inflammation and viral persistence. Persistence of cardiotropic viruses (enterovirus, adenovirus) constitutes one of the predominant aetiological factors in DCM. Additionally, circulating autoantibodies to distinct cardiac autoantigens have been described in patients with DCM, providing evidence for autoimmune involvement. Since clinical complaints of myocarditis and DCM are unspecific, a positive effect of any specific therapy depends on an accurate biopsy-based diagnosis and characterization of the patients with histological, immunohistological and molecular biological methods (PCR), which have developed into sensitive tools for the detection of different viruses, active viral replication, and myocardial inflammation. The immunohistochemical characterization of infiltrates has supported a new era in the diagnosis of myocardial inflammation compared with the Dallas criteria, which has led to a new entity of secondary cardiomyopathies acknowledged by the WHO, the inflammatory cardiomyopathies (DCMi). Immunohistochemically quantified lymphocytes significantly better reflect troponin levels and correlate with findings by anti-myosin scintigraphy compared with the histological analysis. Furthermore, the orchestrated induction of endothelial cell adhesion molecules (CAMs) in 65% of DCM patients has confirmed that CAM induction is a prerequisite for lymphocytic infiltration in DCMi. The combination of these immunohistological with molecular biological diagnostic techniques of virus analysis allows a further classification of dilated cardiomyopathy by differentiating the disease entity in subgroups of virus-positive and virus-negative patients with or without cardiac inflammation. Further analysis of the predominant Th1-/Th2-immune response may provide additional prognostic information on the natural course of the disease. This differential analysis improves the clinical management of patients and is an indispensable prerequisite for the development of specific antiviral or immunomodulatory treatment strategies.

Cardiomyopathies↗

Extrapulmonary pneumocystosis.

Extrapulmonary pneumocystosis is an exceedingly rare complication of Pneumocystis carinii pneumonia (PCP). Prior to the advent of the human immunodeficiency virus type 1 (HIV-1) epidemic, only 16 cases of extrapulmonary pneumocystosis in individuals who were immunocompromised by a variety of underlying diseases had been reported. Since the beginning of the HIV-1 and related PCP epidemic, at least 90 cases of extrapulmonary pneumocystosis have been reported. This review briefly presents a history of the discovery of P. carinii and its recognition as a human pathogen, the controversy regarding its taxonomy, and the epidemiology of this organism. A more detailed analysis of the incidence of extrapulmonary pneumocystosis in HIV-1-infected individuals and its occurrence despite widespread prophylaxis for PCP with either aerosolized pentamidine or systemic dapsone-trimethoprim is presented. The clinical features of published cases of extrapulmonary pneumocystosis in non-HIV-1-infected individuals are summarized and contrasted with those in HIV-1 infected individuals. The diagnosis of extrapulmonary pneumocystosis is discussed, and because clinical microbiologists and pathologists are the key individuals in establishing the diagnosis, the characteristic microscopic morphology of P. carinii as its appears when stained with a variety of stains is presented and reviewed. The review concludes with a brief discussion of treatments for extrapulmonary pneumocystosis.

Classification↗