Pediatric visceral leishmaniasis in Austria: diagnostic difficulties in a non-endemic region.

Visceral leishmaniasis is usually fatal if left untreated. In Europe it is mainly caused by Leishmania infantum which is endemic in the whole Mediterranean region. While visceral leishmaniasis classically affects children, adults increasingly suffer infections in regions which are known to be endemic for HIV. Nowadays up to 70% of the patients with visceral leishmaniasis in southern Europe are HIV-infected adults. The diagnosis is known to be especially difficult to establish in this group of patients because of a frequently atypical clinical presentation, but even in non-HIV-infected patients visceral leishmaniasis often represents a diagnostic challenge particularly when the patient is living in a non-endemic region. We report on four children with visceral leishmaniasis diagnosed at St. Anna Children's Hospital, Vienna, in the last decade. Diagnostic difficulties arose (1) from inexperience with this rare disease, (2) from a long incubation period (6 to 8 months) and (3) from a travel history apparently unsuspicious for the contraction of what is considered a 'tropical' disease. In one case, specific problems resulted (4) from clinical appearance and laboratory data mimicking hemophagocytic lymphohistiocytosis. Consequently even in regions where leishmaniasis is not endemic, diagnostic efforts should be undertaken to rule out this disease especially in patients with the presumptive diagnosis of hemophagocytic lymphohistiocytosis.

[Immunofluorescence and immunocytochemical stain methods for simultaneous cytogenetic and phenotypic characterization of mitotic cells]. / Immunfluoreszenz und immunzytochemische Färbemethoden zur simultanen zytogenetischen und phänotypischen Charakterisierung mitotischer Zellen.

Cytogenetic analysis and immunological phenotyping are well established methods for characterizing leukaemic cell populations. The simultaneous evaluation of cell phenotype and karyotype has, however, been established only recently. We present immunocytochemical methods utilizing horseradish peroxidase and alkaline phosphatase monoclonal anti-alkaline phosphatase staining procedures, as well as modified immunoflurescence methods with fluorescein- and rhodamine-conjugated antibodies. Chromosomes were either stained with Giemsa and "stains-all" (immunocytochemical techniques) or chromomycin and distamycin A/DAPI for R- and C-banding (immunofluorescence techniques), respectively. The advantages and disadvantages of these methods are compared and provide representative examples of the various preparations. These methods will contribute to defining the phenotype of cytogenically-normal cells in leukaemias and to assigning different karyotypes to heterogenous cell populations, as well as supporting the search for analysis of residual blast cells.

Immunoenzymatic staining methods for simultaneous demonstration of chromosomes and cell surface markers.

We have developed immunocytochemical staining methods for the simultaneous phenotypic and karyotypic characterization of individual cells. Following a mild hypotonic pretreatment, isolated cells are cytocentrifuged on poly-L-lysine coated slides, fixed in formol buffered acetone, and subsequently labeled with monoclonal antibodies utilizing indirect immunoenzymatic staining procedures with horseradish peroxidase (HRP) or alkaline phosphatase monoclonal anti-alkaline phosphatase (APAAP) as second antibodies. Preparations are refixed consecutively in methanol and 45% acetic acid and counterstained with either "Stains-all" (HRP labeled preparations) or Giemsa (APAAP labeled preparations). C-banding or weak G-banding, which allows the identification of individual chromosomes, can be induced in labeled as well as unlabeled mitotic cells by Ba(OH)2 and/or 2 X SSC treatment after refixation, respectively. Our method has been successfully tested with a variety of monoclonal antibodies against lymphoid, myeloid, erythropoietic, and thrombopoietic cell surface antigens. It is fast, allows the adjustment of the intensity of cell surface staining, and results in permanent preparations suitable for light microscopic analysis.

[The Netherton syndrome: clinical characteristics, differential diagnosis and new ways of therapy]. / Das Netherton-Syndrom: Klinische Charakteristik, differential-diagnostische Abgrenzung und neue Wege der Therapie.

The Netherton-syndrome is a rare disease which is probably inherited through an autosomal recessive trait. It is defined by a triad of symptoms: congenital ichthyosiform erythrodermia , trichorrhexis invaginata et nodosa ("bamboo hair") and atopy. Additional disorders affect the immune system, the metabolism of amino acids and the physical development. On the basis of a new case, the cellular immune defect and the genetic background of the disease are more clearly defined. A new form of treatment--a combination of photochemotherapy (PUVA) and systematic application of aromatic retinoid--has so far proved to be successful. In order to establish an accurate diagnosis--a prerequisite for this promising therapeutic approach--diseases with similar symptoms are discussed for comparison.