Determination of the donor:host blood cell ratio after haematological stem cell transplant by means of semiquantitative detection of short tandem repeat polymorphisms

An improved method of quantifying the donor:host blood cell ratio after haematological stem cell transplantation (PBSCT) using a variable number of tandem repeat (VNTR) markers is presented. The post-transplant DNA is extracted from the patient's blood and amplified by semiquantitative polymerase chain reaction (PCR) without prior mock transplant and plotting of a standard curve. The amplification products are then analysed by ABI PRISM 310 capillary electrophoresis apparatus. The resultant peak areas are correlated with the corresponding microsatellite allele by GeneScan software and the donor:host cell ratio is calculated. Improvements to PCR amplification conditions are essential for the outcome of the quantification since preferential amplification of alleles in the PCR process can account for the marked deviation found between the results gained by measurement of different microsatellite loci. To assess the accuracy of the method, the post-transplant blood samples of 6 patients who had undergone either myeloablative or non-myeloablative transplantation regimens were analysed retrospectively (median observation time 298 days). By analysing 3 or 4 microsatellite loci we were able to detect full engraftment or mixed chimaerism after transplant with a measurement precision of < or = 4.5 (standard deviation). Sensitivity for different primers ranges from 2% to 5%. The results of the microsatellite analysis correlated well with the corresponding clinical findings. We conclude that post-transplant analysis of microsatellite loci using semiquantitative PCR without standard is suitable for clinical purposes.     

Monoclonal antibody labels olfactory and visual pathways in Drosophila and Apis brains

We employed a monoclonal antibody raised against Drosophila brain homogenate for a comparative immunocytochemical analysis of visual and olfactory pathways in brains of two insect species. On Western blots of Drosophila and Apis nervous tissue, antibody fb45 recognized an antigen with an apparent molecular weight higher than 180 kD. Application of the antibody to sections of Drosophila and Apis brain stained certain interneurons which conspicuously fasciculate in common tracts or neuropilar compartments. Both in Drosophila and in Apis, the antigen was also expressed on the perineural sheath and granular cell compartments in the majority of neuronal cell bodies. The antibody stained monopolar cells in the visual system of both species, and in Apis those fibers of the anterior superior optic tract which link the medulla with the mushroom bodies. In Drosophila, bundles of Kenyon cells of the mushroom bodies were stained. In worker bees and drones, the relay neurons of the median and lateral antennoglomerular tracts were labelled. Since the recognition of the antigen does not require fixation, the antibody can be employed to label selectively living neurons in dissociated cell culture. This opens up the possibility for future functional studies on the role of the antigen in vitro. © 1993 Wiley-Liss, Inc.     

Does early stem-cell transplantation have a role in chronic myeloid leukaemia? For the proposal

Allogeneic haemopoietic stem-cell transplantation (HSCT) can induce long-lasting complete molecular remissions in most patients with chronic myeloid leukemia (CML) for more than 20 years. No other intervention for this disease has such a track record, and early HSCT, despite its associated morbidity and mortality, has become standard care for patients with a compatible donor. This situation has changed with the introduction of the specific tyrosine kinase inhibitor imatinib. The excellent short-term results of imatinib have changed attitudes, and the number of transplantations for CML have declined rapidly since 1999. In this debate, Alois Gratwohl and Gianantonio Rosti present opposing arguments as to whether there is still a role for early HSCT in CML.     

Pumpless extracorporeal lung assist (pECLA) in patients with acute respiratory distress syndrome and severe brain injury

BACKGROUND: A retrospective analysis was performed to estimate the practicability of a pumpless extracorporeal lung assist system (pECLA) in trauma patients suffering from severe brain injury and the acute respiratory distress syndrome (ARDS). METHODS: Five patients with acute severe brain injury and ARDS, ventilated in a lung protective mode, were connected to pECLA to avoid the detrimental effects of hypercapnia on intracranial pressure (ICP) and cerebral outcome. With pECLA hypercapnia was eliminated in all patients while the minute volume of artificial ventilation could be reduced. Subsequently, ICP was reduced, systemic hemodynamics and cerebral perfusion pressure remained stable. One patient died due to multi-organ failure as a consequence of multi-trauma. The remaining patients survived showing a good neurologic function. CONCLUSIONS: pECLA is a promising alternative compared with conventional pump-driven systems for patients with ARDS and brain injury, since the pECLA system has minor restrictions, limitations and side effects.     

Rapidly progressive dysphagia caused by Forestier’s disease: a case report

Summary Forestiers disease is a systemic rheumatologic abnormality of unknown etiology, characterized by a flowing ossification of the anterior ligament of the spine. In this case study we report on an atypical appearance of Forestiers disease in a 72-year-old woman. This patient had a one-month history of rapidly progressing dysphonia and dyspnea and at the time of admission was unable to eat. She had been operated for gastric and colon carcinoma seventeen months earlier. Total body CT scans showed a flowing ossification of the anterior ligament between levels C2 and C7 and an osteophyte protruding in the ventral direction at level C2. Before the osteophytes removal, a tumor screening was conducted to exclude the presence of distant metastases or occult malignancy. Resection of the large osteophyte was performed via a typical ventral-cervical approach with horizontal skin incision. Six months after the procedure, the patient experienced distinct improvement of her condition and was able to eat both pulpy and solid foods again. In this case example, a rapidly progressive dysphagia was not caused by secondary malignancy, as initially speculated, but resulted from a ventral-cervical osteophyte, considered a rare cause for acute development of severe dysphagia. Early diagnosis of Forestiers disease in this patient led to immediate surgical intervention, which proved to be adequate treatment and enabled the patient to make a good recovery.

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Rasch progrediente Dysphagie durch Morbus Forestier: ein Fallbericht

Zusammenfassung Der Morbus Forestier ist eine rheumatologische Systemerkrankung unbekannter Ätiologie, welche durch eine langstreckige Verkalkung des vorderen Längsbandes der Wirbelsäule gekennzeichnet ist. Im Folgenden wird eine atypische Manifestation des Morbus Forestier in einer 72-jährigen Patientin beschrieben. Die Patientin wurde bei seit einem Monat bestehender Dysphonie, Dyspnoe und völliger Unfähigkeit zu schlucken stationär aufgenommen. Siebzehn Monate zuvor wurde die Patientin an einem Magen- und Kolonkarzinom operiert. Das durchgeführte Ganzkörper-CT zeigte eine ausgedehnte Verkalkung des vorderen Längsbandes von C2–C7 mit einem nach ventral ragenden Osteophyten auf Höhe C2. Vor dessen chirurgischer Entfernung wurden mittels eines Tumorscreenings Fernmetastasen oder Lokalrezidive ausgeschlossen. Die Resektion des prominenten Osteophyten erfolgte über einen typischen ventralen Zugang zur Halswirbelsäule. Sechs Monate nach der Operation konnte die Patientin wieder feste und breiige Nahrung aufnehmen. In diesem Fallbeispiel war die rasch progrediente Dysphagie nicht durch sekundäre Absiedelungen, wie initial vermutet, verursacht, sondern durch einen massiven ventralen zervikalen Osteophyten, welcher an sich selten als Ursache für eine akut progrediente komplette Dysphagie vorkommt. Die frühzeitige Diagnose des Morbus Forestier führte unmittelbar zur operativen Dekompression und erwies sich in diesem Fall als adäquate Behandlung mit guter Remission.

     

Recombinant protein and DNA vaccines derived from hsp60 Trichophyton mentagrophytes control the clinical course of trichophytosis in bovine species and guinea-pigs

Heat shock proteins (hsp) were identified in many infectious agents as immunodominant antigens with a protective effect. Immunization of laboratory animals by selected representants of hsp60, hsp70 and hsp90 isolated from several pathogens induced protective host immunity and significantly reduced clinical manifestation of infection. The present study involves preparation of a recombinant protein vaccine and a DNA vaccine. Both vaccines were derived from the hsp60 of the dermatophyte, Trichophyton mentagrophytes. Challenge trials with evaluation of the protective effect of vaccination were performed on calves and guinea-pigs. Both vaccination procedures reduced, in statistically significant fashion, the clinical course of skin mycosis in calves experimentally inoculated with the dermatophyte, T. mentagrophytes. In experiments with guinea-pigs, increased protection was only seen with DNA vaccination. After DNA vaccine application, no paravaccination side-effects (granulomas at the injection site, changed total state of the animal) were observed. Only vaccination with the recombinant protein in calf's experiment induced specific serum antibodies. This observation indicates that antibodies are not associated with protection. In summary, DNA vaccine hsp60 is the most promising for prevention of bovine trichophytosis.     

Drug history overrides opioid reinforcement in a rat runway procedure

To determine the effect of the drug history of rats on their subsequent operant behaviour in the rat runway procedure, we tested the mu opioid receptor agonists morphine and remifentanil and found a carryover of previous drug/saline experience that was not found for a food reinforcer. Previous exposure to saline significantly decreased the apparent reinforcing effect of subsequently offered morphine or remifentanil, while previous experience with morphine or remifentanil significantly increased responding for saline. This carryover of a previous learning experience on subsequent operant performance in the rat runway procedure cautions that the drug history should be considered and might even invalidate subsequent findings when testing drugs of abuse or other reinforcers.     

Induction of aryl hydrocarbon receptor-mediated and estrogen receptor-mediated activities, and modulation of cell proliferation by dinaphthofurans

A group of heterocyclic aromatic compounds, dinaphthofurans (DNFs), recently have been identified as potentially significant contaminants in freshwater sediments. In the present study, a battery of in vitro assays was used for detection of toxic effects of DNFs that are potentially associated with endocrine disruption and tumor promotion. Dinaphthofurans were found to act as relatively potent inducers of aryl hydrocarbon receptor (AhR)-mediated activity in the chemical-activated luciferase reporter gene expression DR-CALUX assay. The relative AhR-inducing potencies of DNFs were similar or even higher than relative potencies of unsubstituted polycyclic aromatic hydrocarbons (PAHs), with dinaphtho[1,2-b;2'3'-d]furan being the most potent AhR agonist. Two compounds, dinaphtho[2,1-b;2'3'-d]furan and dinaphtho[1,2-b;1'2'-d]furan, induced estrogen receptor (ER)-mediated activity in the estrogen receptor-mediated CALUX (the ER-CALUX) assay. Two types of potential tumor-promoting effects of DNFs were investigated, using in vitro bioassays for detection of inhibition of gap-junctional intercellular communication and detection of a release from contact inhibition. Although the acute inhibition of gap-junctional intercellular communication was not observed, all six tested DNFs were able to release rat liver epithelial WB-F344 cells from contact inhibition at concentrations as low as 100 nM. In summary, the present study indicated that DNFs can exert multiple biological effects in vitro, including induction of the AhR-mediated activity, release of cells from contact inhibition, and induction of ER-mediated activity.