Isolation and immunoelectronmicroscopical characterization ofTheileria annulata macroschizonts

Theileria annulata macroschizonts were isolated from bovine lymphoblastoid cells grown in cell culture. To release the parasites, the cells were homogenized under hypotonic conditions. Intact host lymphocyte nuclei were lysed and the resulting chromatin precipitate was degraded by DNase. Host cell fragments were removed by ion-exchange chromatography. As revealed by electron microscopy, the preparations were free of intact host lymphocytes, lymphocyte nuclei and organelles. Antisera raised in rabbits against purified macroschizonts showed a specific reaction with the intracellular parasite in the indirect immunofluorescence test and in immuno-electron microscopy.     

High Throughput Fingerprint Analysis of Large-Insert Clones

As part of the Human Genome Project, the Washington University Genome Sequencing Center has commenced systematic sequencing of human chromsome 7. To organize and supply the effort, we have undertaken the construction of sequence-ready physical maps for defined chromosomal intervals. Map construction is a serial process composed of three main activities. First, candidate STS-positive large-insert PAC and BAC clones are identified. Next, these candidate clones are subjected to fingerprint analysis. Finally, the fingerprint data are used to assemble sequence-ready maps. The fingerprinting method we have devised is key to the success of the overall approach. We present here the details of the method and show that the fingerprints are of sufficient quality to permit the construction of megabase-size contigs in defined regions of the human genome. We anticipate that the high throughput and precision characteristic of our fingerprinting method will make it of general utility.     

Extraskeletal myxoid chondrosarcoma: multimodal diagnosis and identification of a new cytogenetic subgroup characterized by t(9;17)(q22;q11)

Extraskeletal myxoid chondrosarcoma is a rare malignant soft tissue tumour that can be difficult to diagnose correctly, especially preoperatively. We describe four cases of extraskeletal myxoid chondrosarcoma of the extremities diagnosed by a multimodal approach. The cytological examination of fine-needle aspirates showed small and round, mildly pleomorphic cells lying in sheets and cords, but also dispersed within a myxoid and metachromatic intercellular substance. Histological, electron microscopic and immunocytochemical examination also yielded findings compatible with the diagnosis of extraskeletal myxoid chondrosarcoma. Cytogenetic analysis demonstrated a t(9;22)(q22;q12) in two tumours and a t(9;17)(q22;q11) in the third and fourth. The translocation t(9;22)(q22;q12) has been described repeatedly in extraskeletal myxoid chondrosarcoma but never in other tumours; hence, the detection of this pathognomonic chromosome abnormality in short-term cultured cells from fine-needle aspirates verified the diagnosis in two of the cases. The t(9;17)(q22;q11) found in the last two cases probably represents a new cytogenetic subgroup of extraskeletal myxoid chondrosarcoma as it, too, is unknown in other contexts. The multimodal approach taken in these four cases enabled a definite diagnosis of a rare malignant tumour whose cytological and histological features alone are usually not sufficiently distinct to rule out other differential diagnostic possibilities. Received: 16 March 1999 / Accepted: 1 June 1999     

Vergleich zwischen cytologischer und cytochemischer Klassifizierung bei 47 Fällen von akuter Leukämie

Zusammenfassung Bei 47 Patienten mit akuter Leukämie wurden die Ergebnisse einer voneinander unabhängigen Klassifizierung aufgrund der panoptischen Färbung einerseits und dem Ausfall cytochemischer Reaktionen andererseits (Peroxydase, PAS-Reaktion, Naphthol-AS-Acetat-Esterase) sowie das Ansprechen auf eine adäquate Induktionstherapie in den cytologisch und cytochemisch definierten Gruppen miteinander verglichen. In 42 Fällen stimmte die cytologische und cytochemische Differenzierung miteinander überein. Hinsichtlich des Ansprechens auf Therapie ergeben sich keine signifikanten Unterschiede, gleichgültig, ob man die AL nach cytologischen oder cytochemischen Kriterien gruppiert. Eine Ausnahme bildet der bei undifferenzierten AL cytochemisch charakterisierbare PAS-Typ, bei dem ein besseres Ansprechen auf Therapie als bei den Fällen ohne diagnostisch verwendbare cytochemische Aktivität wie auch bei den granulocytär differenzierten AL gefunden wurde. Hinsichtlich des Ansprechens auf Therapie verhalten sich die Erwachsenen mit PAS-Typ ähnlich günstig wie die Kinder.Mit Unterstützung der Deutschen Forschungsgemeinschaft.     

Effects of sex steroid analogs on spontaneous autoimmune thyroiditis in obese strain chickens.

Treatment of obese strain chickens, either in ovo or after hatching, with testosterone or various testosterone-derived steroid analogs having no or only weak endocrine activities causes significant protection from spontaneous autoimmune thyroiditis occurring in these birds. This effect is not correlated with the endocrinological properties of these steroids. This observation and similar results in other models for autoimmunity may present a basis for a more selective therapy in human autoimmune diseases in that hormonal side effects no longer preclude this approach.     

Polymerisationsauslösung mit substituierten ethanen, 2. Trennung der Oligomeren aus Methylmethacrylat und 1,1,2,2-Tetraphenyl-1,2-diphenoxyethan

The telechelics 1 from methyl methacrylate and 1,1,2,2-tetraphenyl-1,2-diphenoxyethane, can be separated by adsorption chromatography. The structure of the dimers and trimers was identified by ¹H NMR-spectroscopy. The free radical oligomerization is a process in which the syndiotactic structure is favoured above the isotactic one.     

Quantitative reflection spectroscopy at the human ocular fundus

A new model of the reflection of the human ocular fundus on the basis of the adding-doubling method, an approximate solution of the radiative transport equation, is described. This model enables the calculation of the concentrations of xanthophyll in the retina, of melanin in the retinal pigment epithelium and the choroid, and of haemoglobin in the choroid from fundus reflection spectra. The concentration values found in 12 healthy subjects are in excellent agreement with published data. In individual cases of pathologic fundus alterations, possible benefits to the ophthalmologic diagnostics are demonstrated.     

Dosimetric characteristics of a new unshielded silicon diode and its application in clinical photon and electron beams

Shielded p-silicon diodes, frequently applied in general photon-beam dosimetry, show certain imperfections when applied in the small photon fields occurring in stereotactic or intensity modulated radiotherapy (IMRT), in electron beams and in the buildup region of photon beam dose distributions. Using as a study object the shielded p-silicon diode PTW 60008, well known for its reliable performance in general photon dosimetry, we have identified these imperfections as effects of electron scattering at the metallic parts of the shielding. In order to overcome these difficulties a new, unshielded diode PTW 60012 has been designed and manufactured by PTW Freiburg. By comparison with reference detectors, such as thimble and plane-parallel ionization chambers and a diamond detector, we could show the absence of these imperfections. An excellent performance of the new unshielded diode for the special dosimetric tasks in small photon fields, electron beams and build-up regions of photon beams has been observed. The new diode also has an improved angular response. However, due to its over-response to low-energy scattered photons, its recommended range of use does not include output factor measurements in large photon fields, although this effect can be compensated by a thin auxiliary lead shield.     

DNA cross-link-dependent RAD50/MRE11/NBS1 subnuclear assembly requires the Fanconi anemia C protein

Fanconi anemia (FA) is a cancer-predisposition syndrome characterized by hypersensitivity to interstrand-cross-link (ICL) inducers. FA hypersensitivity to ICL has been correlated with alterations in homologous recombination, non-homologous end-joining, telomere maintenance, DNA-damage assessment and checkpoint regulation, processes in which the components of the RAD50/MRE11/NBS1 (RMN) complex are involved. To better characterize the mechanisms by which ICL are processed in human cells and to gain insight into their toxicity in FA, we examined (i). the RMN complex assembling in response to the ICL inducers mitomycin C (MMC) and photoactivated 8-methoxypsoralen and (ii). the proficiency of FA cells to perform RMN activation in response to ICL inducers. We show here that ICL activates the assembly of the RMN proteins into subnuclear foci, and that their formation proceeds independently of ICL incision, a step mainly dependent on XP-F/ERCC1 heterodimer activity. Interestingly, FA cells were unable to form RMN foci in response to either ICL inducer. Analysis by pulsed-field gel electrophoresis and single-cell gel electrophoresis of MMC-treated cells showed that FA cells from complementation group C (FA-C cells, defective in the FANCC gene) form double-strand breaks and unhook MMC-induced ICL similarly to FANCC wild-type cells. These observations imply that the absence of RMN assembly in FA-C cells is not simply due to the absence of DNA ends produced as intermediates of ICL processing, and indicates a direct role for FANCC in RMN focus assembly in response to ICL inducers. Moreover, we show that the formation of foci, including BRCA1 and/or RAD51 proteins, is significantly delayed in FA cells. These alterations in the assembly of DNA-repair proteins in FA provide an interpretation for the DNA-damage processing anomalies observed in FA cells and for the genetic instability and the cancer predisposition of the syndrome.